Venkatachalam/aging_reg_dataset · Datasets at Hugging Face

WIF1

11197

protein coding

PA116,CaExPA79

Primary salivary gland tumor

Salivary gland tumor

Accelerate

Cell cycle analysis//Cell proliferation assay//SA--gal activity assay

Re-expression of WIF1 resulted in a significant growth inhibition (P<0.0001) in both PA and CaExPA cells at all time points.Cells were stained for senescence-associated beta-galactosidase (SA-¦Â-gal). WIF1 increased the number of SA-¦Â-gal-positive cells (P<0.0001) compared with vector.

p53//p21

Upregulation//Upregulation

RT-PCR//Western blot

?WIF1 induced a remarkable increase in both mRNA and protein expression of p53 and its target p21.

--

Human

L

cellular senescence

24,853,424

Gene

0

1

0

1

0

1

0

MUC4

4585

protein coding

UMSCC10B,UMSCC1

--

Squamous cell carcinoma

Prevent

Cell cycle analysis//Growth kinetics analysis//SA--gal activity assay

A significant decrease in growth rates of MUC4 KD SCC1 and SCC10B cells was observed indicating the role of MUC4 in cell proliferation.;Our SA-¦Â-gal staining showed 37¡À4% and 27¡À6% positive in MUC4 KD SCC1 and SCC10B cells compared to 3¡À2% and 4¡À1% in control cells respectively.

--

p16-Rb

--

Western blot//SA-¦Â-gal activity assay

We observed significant upregulation of p16 protein and downregulation of cyclin E, cyclin D1, decreased phosphorylation of pRb in MUC4 KD cells compared to control cells.

Human

L

cellular senescence

24,747,969

Gene

0

1

0

1

0

1

0

NOL12

79159

protein coding

HDF

Skin

Aging

Prevent

BrdU assay//Flow cytometry//SA--gal activity assay

Proliferation rate of NOL12-depleted cell cultures, confirmed by a decreased percentage of living cells that incorporated 5-ethynyl-2¡ä-deoxyuridine (EdU), with cycling cells from NOL12-depleted cell cultures exhibiting a cell cycle delay in comparison to the cycling of the control£»enescence-associated ¦Â-Gal positive .

--

p53

--

Immunostaining//Western blot//Knockdown

Both immunoblot and immunofluorescence quantitative analyses of p53 levels revealed its stabilization upon NOL12 and XRN2 knockdowns?; RPL11 to be required for p53 activation upon NOL12 knockdown. upon NOL12 knockdown, we detected a significant induction of the p53 downstream target p21/CDKN1A.

Human

L

cellular senescence

30,988,155

Gene

0

1

0

1

0

1

0

MEOX2

4223

protein coding

IMR-90,WI-38,HEKn

--

Aging

Accelerate

FACS analysis//SA--gal activity assay

MEOX2 -expressing cells exhibited reduced proliferative capability and progressive cell size increase,£»Fibroblasts with forced MEOX2 expression also harbored significantly higher senescence-associated Beta-galactosidase activity£¬MEOX2 expression also led to an accumulation of cells in the G1 phase of the cell cycle, and a concomitant decrease of cells in S phase, consistent with cells undergoing senescence.

INK4a

Upregulation

Western blot//IP

NK4a protein levels were increased two-fold by MEOX2 overexpression in fibroblasts .Immunoprecipitation of MEOX2-HA strongly enriched for sequences located within the INK4a promoter region.

--

Human

L

cellular senescence

19,340,300

Gene

0

1

0

1

0

ERCC6

2074

protein coding

IMR-90

--

Aging

Prevent

Knockdown//SA--gal activity assay

Proliferation arrest of CSB-knocked down fibroblasts earlier (PN26£¬ PN24) compared to cells transduced with shSCR (PN28). Precocious senescence upon transient CSB knockdown was confirmed by a higher number of SA-¦Â-gal+ cells.

p21

Downregulation

CHIP//qRT-PCR

CSB directly interacts with two regions of the?p21Waf1?promoter that are target of p5329, and a more extended interaction in the distal (5¡¯) region was observed with the C-terminal anti-CSB than with N-terminal anti-CSB.

DDR-p21

--

Immunostaining//Western blot//SA-¦Â-gal activity assay

Decreased CSB levels upon irradiation resulted in the loss of CSB interaction with the p21Waf1 promoter £»Senescence following irradiation was verified by high levels of the DDR-dependent senescence marker p21.

Human

HL

cellular senescence

31,811,121

Gene

0

1

0

1

0

SPOP

8405

protein coding

IMR-90

--

Prostate cancer

Accelerate

BrdU assay//Knockdown//SA--gal activity assay//Western blot

Ectopic SPOP expression induced expression of markers of senescence such as SA-¦Â-gal activity and upregulation of p53 and p21, while it suppressed cell proliferation markers .SPOP knockdown suppressed the expression of senescence markers such as SA-¦Â-gal activity, SAHF formation and upregulation of p53 and p21.cell proliferation markers were significantly increased in shSPOP/RAS cells compared with RAS-expressing cells.

SENP7//HP1¦Á

Downregulation//--

Co-IP//Immunohistochemistry//Knockdown

Ectopic SPOP and SENP7 co-immunoprecipitated with each other;here was a SPOP dose-dependent decrease in SENP7 expression , suggesting that SPOP promotes SENP7 degradation.; SENP7 is expressed at significantly (p<0.0001) higher levels in prostate tumors with SPOP mutations compared to those with wild-type SPOP.SPOP knockdown caused a decrease in the levels of sumoylated HP1¦Á in senescent cells.

--

Human

L

cellular senescence

26,527,005

Gene

0

1

0

1

0

1

0

1

0

SRSF1

6426

protein coding

Primary Fibroblast

--

Aging

Accelerate

Cell proliferation assay//EdU assay//SA--gal activity assay

SRSF1 overexpression resulted in a 5-fold increase in the number of SA-¦Â-gal-stained cells;Prolonged overexpression of SRSF1 resulted in enlarged and flattened cells with an increased number of intracellular vesicles¡ªphenotypes typical of senescence;Within four days of SRSF1 overexpression, cell proliferation was drastically reduced to only 20% of control cells.

RPL5//p53

--//--

RT-PCR//Western blot//Knockdown

siRNA-mediated knockdown of RPL5 in BJ cells severely abrogated the effect of SRSF1 overexpression on p53.

--

Human

L

cellular senescence

23,478,443

Gene

0

1

0

1

0

1

0

NOLC1

9221

protein coding

2BS

--

Hepatocellular carcinoma

Accelerate

Cell cycle analysis//Knockdown//SA--gal activity assay

NOLC1 overexpression could increase the activity of SA©\¦Â©\gal, and knockdown of NOLC1 repressed the enhanced SA©\¦Â©\gal activity induced by CSIG knockdown£»the overexpression of NOLC1 repressed the cell proliferation and promoted cell cycle arrest in 2BS cells.

CSIG

--

Western blot//qRT-PCR//Knockdown

CSIG knockdown increased the expression of NOLC1£»the overexpression of CSIG could clearly repress the expression of NOLC1.

--

Human

HL

cellular senescence

28,493,459

Gene

0

1

0

1

0

1

0

HDAC1

3065

protein coding

HeLa

--

Cervical cancer

Accelerate

Cell morphological analysis//SA--gal activity assay//Western blot

Some characteristics of senescence, including an enlarged and flattened morphology, positive staining for SA-¦Â-galactosidase activityand increased autofluorescence, were observed in HDAC1 over-expressing cells;cells expressing high levels of HDAC1 immediately suppressed cell division.

--

Sp1-PP2A-pRb

--

Western blot//Luciferase reporter assay//Chromatin-immunoprecipitation assay

An approximately 3-fold induction increase in the activity of the PP2Ac promoter in the presence of Sp1;HDAC1-overexpressing cells exhibited a dose-dependent inhibition of pRb1;HDAC1 activated the promoter activity of PP2Ac and increased protein expression of PP2Ac.

Human

L

cellular senescence

21,420,382

Gene

0

1

0

1

0

1

0

TERT

7015

protein coding

RPE

--

Aging

Prevent

SA--gal activity assay

We stained hTRT-RPE clones at or near senescence and compared the level of SA¨C¦Â-Gal staining to that in hTRT+clones that had undergone a similar or greater number of cell divisions. A majority of the cells in the hTRT-clones showed strong staining; by contrast, few of the cells in hTRT+clones at equivalent or greater PD showed staining.

--

Human

L

cellular senescence

9,454,332

Gene

0

1

0

HELLS

3070

protein coding

2BS,WI-38

--

Aging

Prevent

Cell growth assay//Flow cytometry//Knockdown//SA--gal activity assay

Nearly all of the Lsh¨CshRNA-infected cells (PD 42) showed strong levels of blue SA-¦Â-gal staining similar to senescent cells;Lsh blocks the formation of senescence-associated heterochromatin foci (SAHF); growth of Lsh-HA-transfected cells advanced quickly, suggesting a strong proliferative potential relative to the control cells . In contrast to the severe G1 cell-cycle arrest imposed by Lsh-shRNA, the control-shRNA cells showed a decrease in the proportion of 2BS cells in the G0/G1 phases.

p16//HDAC

Downregulation//--

Western blot//RT-PCR//Pull-down assay//IP

A marked increase in p16?INK4a?protein levels in Lsh-shRNA cells comparing to control-shRNA cells.Lsh-shRNA-infected cells showed a significant increase in p16 mRNA levels in 2BS cells .The results showed that Lsh immunoprecipitates from Lsh-deficient cells contained significantly decreased the reverse immunoprecipi- tation displayed the similar result. In addition, GST pull-down experiments with bacteria expressed GST-Lsh and in vitro transcripted/translated HDAC1 were then performed to examine the nature of the interaction between Lsh and HDAC1. HDAC1 levels relative the mock-transfected cells.

--

Human

L

cellular senescence

19,561,196

Gene

0

1

0

1

0

1

0

1

0

TERF2

7014

protein coding

U2OS

--

Aging

Prevent

PI staining//SA--gal activity assay

We analyzed cell cycle profiles in TRF2-depleted cells and found an increase in the proportion of cells in the G2 phase of cell cycle . following addition of DOX ,the number of TRF2-depleted cells compared with control uninduced cells progressively decreases,inducible clones became strongly positive for acidic ¦Âgal (SA-¦Âgal), a marker of cellular senescence.

p53

--

Western blot

TRF2-depleted cells (G10) p53 is phosphorylated and its levels increased.

--

Human

L

cellular senescence

18,056,407

Gene

0

1

0

1

0

PML

5371

protein coding

IMR-90

--

Prostate cancer

Accelerate

Crystal violet assay//SA--gal activity assay

PML-expressing fibroblasts arrested their proliferation rapidly after selection and developed characteristics of cellular senescence, including a flat morphology, positive staining for the senescence-associated ¦Â-galactosidase (SA-¦Â-Gal), expression of high levels of IL-8,and markers of mitochondrial dysfunction and biogenesis.

E2F//p53

Downregulation//--

CHIP//qPCR

Several E2F target genes that play a role in DNA replication, cell cycle progression, and DNA repair were found to be down-regulated in normal fibroblasts expressing PML.E2F targets were enriched sixfold among PML down-regulated genes in comparison with a random set of genes;The p53-dependant gene GADD45¦Á was significantly up-regulated 10 d after PML expression.

--

Human

HL

cellular senescence

21,205,865

Gene

0

1

0

1

0

TPR

7175

protein coding

HeLa

--

Cancer

Prevent

Colony formation assay//FACS analysis//Knockdown//SA--gal activity assay

Treatment with Tpr siRNAs impaired cell proliferation as compared to mock siRNA-treated cells.The cell cycle was arrested at the G0¨CG1 phase in Tpr knockdown cells A substantial decrease (36%) in the number of colonies was apparent when cells were transfected with Tpr siRNAs.most of the enlarged and flattened cells were ¦Â-gal positive and colored blue (about 40%) in Tpr knockdown wells.

--

p53

--

Knockdown//Western blot

Levels of p53 and p21 were both up-regulated in Tpr-depleted Hela cells and U2OS cells .Knocking down both Tpr and p53 resulted in a substantial reversion of the senescence phenotype as assessed by the percentage of SA-¦Â-galactosidase-positive cells.

Human

L

cellular senescence

21,811,608

Gene

0

1

0

1

0

1

0

1

0

USP1

7398

protein coding

WI-38,BJ

--

Aging

Prevent

SA--gal activity assay//Western blot//qRT-PCR

We confirmed that oncogenic RAS senescent cells have reduced USP1 transcript and protein levels compared to pre-senescent cells .Using of shUSP1-1, shUSP1-2, and shUSP1-3 induced a cell-cycle arrest with senescence features, including a perma-nent proliferative arrest,a flat cell morphology, and increased senescence-associated beta-galactosidase (SABG)positivity with a concomitant decrease in Ki67 expression.

KDM4A

--

Human

HL

cellular senescence

27,160,904

Gene

0

1

0

1

0

1

0

NEDD4

4734

protein coding

IMR-90,2BS

--

Aging

Prevent

SA--gal activity assay

Nearly all of the NEDD4-1-shRNA-infected cells displayed stronger levels of blue SA-¦Â-gal staining similar to senescent cells, the control cells showed a lower frequency of SA-¦Â-gal staining. However, only sporadic SA-¦Â-gal-positive cells were seen in NEDD4-1 -transfected cells, although the corresponding vector control cells showed strong SA-¦Â-gal staining.

PPAR¦Ã

Downregulation

Western blot

A high level of NEDD4-1 overexpression also caused a decrease of endogenous PPAR¦Ã protein in the Hela cells. Such a decrease of PPAR¦Ã protein level can be blocked by proteasomal inhibitor MG132£»we demonstrated that endogenous NEDD4-1 negatively regulates PPAR¦Ã levels quantitatively following increased RNAi concentration.

--

Human

L

cellular senescence

23,549,616

Gene

0

1

0

UBC

7316

protein coding

BM-MSC

--

Aging

Prevent

Flow cytometry//Knockdown//SA--gal activity assay

SA-¦Â-gal-positive senescent cells were accumulated as 4.7¡À1.9%, 7.8¡À1.9% and 13.7 ¡À5.1% at 24, 48 and 72 h after UBC KD, respectively,while they were not increased in NC-transfected cells (2.6 ¡À1.9%, 2.6¡À1.3% and 0.8¡À0.2% after 24, 48 and 72 h, respectively);The impaired proliferation of hBM-MSCs occurred in a UBC dose-dependent manner.

hub

--

qRT-PCR

The hub molecules including CDK1, CCNA2, MCM10, E2F1, HIST1H1A, HIST1H3B and BRCA1 were sequentially decreased during passaging accompanied by the decrement of UBC.

--

Human

HL

cellular senescence

29,382,826

Gene

0

1

0

1

0

1

0

PML

5371

protein coding

MRC-5,WI-38

--

Aging

Prevent

Knockdown//SA--gal activity assay

PML knockdown caused a dose-dependent decrease in cell proliferation, morphological changes consistent with a senescent phenotype and an increased expression of the senescence-related ¦Â-galactosidase enzyme.

PML/RAR¦Á//p53//p16//p21

--//--//--//--

Western blot

However, in PML/RAR¦Á expressing HPCs colocalization was only found in 15% of the cells. Moreover, 50% of the control cells had >10% of the PML signals colocalizing with telomeres whereas this was observed in only 3% of the PML/RAR¦Á cells.An increased expression of the p53, p16INK4a?and p21WAF1?proteins in PML-depleted cells with respect to control cells.

--

Human

L

cellular senescence

26,119,943

Gene

0

1

0

1

0

MET

4233

protein coding

MDCK epithelial cell

--

Tumor

Accelerate

BrdU assay//Cell cycle analysis//SA--gal activity assay

RON c5 and c8 were found to be largely arrested in the G0/G1?phase, in comparison to the broader cell-cycle distribution of MDCK and RON c1.BrdU staining indicated that RON c5 and c8 had significantly lower BrdU incorporation compared to that of MDCK and RONc1£»RON c5 and c8 were undergoing senescence, in contrast to that of parental MDCK and RON c1 .

--

Human

L

cellular senescence

17,588,532

Gene

0

1

0

1

0

1

0

PRKCA

5578

protein coding

HEK293,HCT116

--

Aging

Prevent

SA--gal activity assay

Inhibition of cPKC or aPKC resulted in increased SA-¦Â-gal staining as compared with control.

--

Akt-FOXO3a-ROS-p53-p21

--

Immunostaining//Flow cytometry//Western blot

Treatment with triciribine markedly abolished PKC inhibition-mediated nuclear exportation of FoxO3a£»We observed an increase in ROS accumulation, as shown by the rightward shift in DCF and ETH fluorescence in cells due to PKC downregulation£»Accumulation of p53 and p21Cip1/WAF1 proteins was also observed in cells treated with G?o6983 or Ad-DN-PKCz, as compared with untreated control cells, suggesting that downregulation of cPKC and aPKC induces senescence through stabilization of p53 and transcriptional activation of p21Cip1/WAF1.

Human

L

cellular senescence

28,989,024

Gene

0

1

0

SIRT6

51548

protein coding

WI-38

--

Aging

Prevent

Knockdown//SA--gal activity assay

SIRT6?knockdown (S6KD) cells have a strikingly shortened replicative lifespan, undergoing premature cellular senescence about ten population doublings before control cells, and show increased levels of senescence-associated ¦Â-galactosidase (SA-¦Â-gal) staining.

WRN

--

Western blot//Flow cytometry

In both U2OS and IMR90 cells, SIRT6 knockdown significantly inhibited the association of WRN with telomeric chromatin.

--

Human

L

cellular senescence

18,337,721

Gene

0

1

0

1

0

PML

5371

protein coding

IMR-90

--

Aging

Accelerate

BrdU assay//SA--gal activity assay

Cells arrested proliferation shortly after selection and remained arrested with features of cellular senescence, that is, a flat cell morphology and positive staining for the senescenceassociated ¦Â-galactosidase;PML induced a growth arrest with the characteristics of cellular senescence.

--

Rb

--

SA-¦Â-gal activity assay//3H-thymidine incorporation assay

Cells expressing this mutant entered senescence in response to PML with higher efficiency than wild-type HDFs, according to a3 H-thymidine incorporation assay that measures cell cycle arrest, and according to the SA-¦Â-gal assay that measures senescence,Therefore, the ability of E7 to by pass PML-induced senescence requires inactivation of the Rb tumor-suppressor pathway.

Human

L

cellular senescence

14,712,214

Gene

0

1

0

1

0

HMGB1

3146

protein coding

IMR-90,HCA2,HMEC,MEF

Embryo

Aging

Prevent

Knockdown//SA--gal activity assay//Western blot

We depleted HMGB1 in human fibroblasts (IMR-90, HCA2) that reduced intracellular HMGB1 protein levels by >90%. HMGB1-depleted cells underwent senescence, as judged by arrested growth, senescence-associated heterochromatin foci (SAHF) formation (Narita et al., 2003), and SA-¦Â-Gal activity.HMGB1 depletion also caused senescence in human mammary epithelial cells (HMECs) and MEFs, indicating the response was not restricted to human fibroblasts.

p53//NF-¦ÊB

--//--

SA-¦Â-gal activity assay//Western blot//ELISA assay

Consistent with the IL-6 ELISA results, NF-¦ÊB activity was robust in HMGB1-overexpressing cells, but not HMGB1-depleted cells,senescence caused by HMGB1 overexpression or depletion was p53 dependent, as determined by SA-¦Â-Gal activity, growth arrest (not depicted), and SAHF formation.

--

Human

L

cellular senescence

23,649,808

Gene

0

1

0

1

0

1

0

CCN1

3491

protein coding

BJ

--

Aging

Accelerate

BrdU assay//Immunostaining//SA--gal activity assay

Plication of CCN1 arrested cell proliferation within 3 days, as shown by stag-nant cell numbers and >50% decrease in 5-bromodeoxyuridine (BrdU)incorporation and Ki-67-positive cells when compared with bovine serum albumin (BSA)-treated controls ,CCN1-treated fibroblasts exhibited an enlarged and flattened cell morphology characteristic of senescent cells, and fourth, these cells expressed markers of senescence including SA-¦Â-gal, p53 and p16INK4a.

p53//NOX1//p38MAPK//ERK

Activation//Upregulation//Activation//Activation

Western blot//Knockdown//RT-PCR

Both p53 and p16INK4a proteins accumulated on CCN1 treatment, indicating their activation£»CCN1 upregulated NOX1 expression in BJ cells,Knockdown of NOX1 by either of two shRNAs (#1 and #2) partially bypassed CCN1-induced growth arrest.Both kinases were activated by CCN1 in a biphasic manner (3 h and 24 h), and apocynin effectively abrogated ERK and p38 MAPK activation at both early and late phases

p16-pRb//Integrin

--//--

Western blot//Knockdown//SA-¦Â-gal activity assay

Both p53 and p16INK4a proteins accumulated on CCN1 treatment, indicating their activation£»silencing of p16INK4a by either expression of shRNA (shp16INK4a) or its suppressor Bmi-1 partially restored CCN1-induced growth suppression .Pretreatment of cells with either soluble heparin or a function-blocking monoclonal antibody against ¦Á6 integrin inhibited CCN1-induced SA-¦Â-gal expression, whereas an antibody against ¦Áv¦Â3 integrin had no effect, indicating the involvement of ¦Á6¦Â1 integrin and HSPGs. Furthermore, the addition of a peptide that competitively binds ¦Á6¦Â1 integrin specifically blocked CCN1-induced senescence, whereas a peptide with a two-residue substitution (T1-mut) that abrogated binding of ¦Á6¦Â1 integrin was ineffective .

Human

L

cellular senescence

20,526,329

Gene

0

1

0

1

0

1

0

TLR2

7097

protein coding

IMR-90

--

Aging

Accelerate

Crystal violet assay//SA--gal activity assay

Overexpression of TLR2 induced cell cycle arrest and an increase in the number of senescence-associated ¦Â-galactosidase (SA-¦Â-Gal)¨Cpositive cells.

p21//p16//p15INK4b//p53

--//--//--//--

Western blot

?Suppression of TLR2 and TLR10 resulted in a decrease in p21CIP1, p16INK4a, and p15INK4b?mRNA expression and a reduction of p53 protein levels.

NF-¦ÊB//p38 MAPK//A-SAA

--//--//--

Western blot//qRT-PCR

A significant enrichment in genes regulated by NF-¦ÊB in OIS (22) was observed in the transcriptome of control cells compared to TLR2-depleted cells £»We also observed a marked decrease in p38 MAPK phosphorylation in TLR2- and TLR10-depleted cells in OIS .Western blot of the conditioned medium from IMR90 ER:RAS cells showed an accumulation of A-SAAs, which was reduced following siRNA knockdown of SAA1 and SAA2, suggesting that A-SAAs are components of the SASP.Further mRNA analysis by quantitative reverse transcription polymerase chain reaction (qRT-PCR) of AA1, SAA2, SERPINA3, PTGS2, STAT3, and IL-6R expression con-firmed these results, indicating that the expression of A-SAAs is highly induced during OIS and is dependent on TLR2 and TLR10.

Human

HL

cellular senescence

31,183,403

Gene

0

1

0

1

0

GOT2

2806

protein coding

8988T

--

Pancreatic ductal adenocarcinoma

Prevent

Knockdown//SA--gal activity assay

GOT2 knockdown significantly elicited senescence in PDAC cells and also resulted in morphological changes, impaired PDAC growth and accumulation of cells in the G1 phase. Next, we found that overexpression of human GOT2 in PDAC cells can rescue the induction of senescence by GOT2 knockdown.

p27

--

Knockdown//Western blot

When GOT2 was knocked down, p27 mRNA and protein levels were robustly induced in PDAC cells£»to directly test whether p27 is required for the GOT2 knockdown-mediated senescence, we suppressed p27 expression using short interfering RNA (siRNA) in 8988T cells and assessed senescence upon GOT2 knockdown. Remarkably, silencing of p27 almost completely diminished senescence induction.

--

Human

L

cellular senescence

29,352,139

Gene

0

1

0

1

0

PDGFB

5155

protein coding

HDF

--

Aging

Accelerate

SA--gal activity assay//SAHF

Indeed they displayed a flattened morphology , an increase Senescence Associated ¦Â Galactosidase (SA-¦Â-Gal) activity and an increase Senescence associated Heterochromatin Foci (SAHF).

p53

--

Knockdown//SA-¦Â-gal activity assay//SAHF

p53 knockdown strongly decreased the appearance of the senescence population in the PDGFB expressing cells as measured by loss of the SA-¦Â-Gal positive cells and the decreased SAHF positive cells.

--

Human

L

cellular senescence

23,934,686

Gene

0

1

0

1

0

AR

367

protein coding

PC-3

--

Prostate cancer

Accelerate

Cell cycle analysis//SA--gal activity assay

Longterm AR activation resulted in G1 growth arrest; the cells assumed flattened vacuolized morphology, suggestive of either autophagy or senescence. The levels of the principal autophagy mediator Beclin-1 [23,28] remained stable in the presence of DHT pointing to senescence. SA-¦Â-Gal positivity increased from the background 4% to nearly 40% in the presence of DHT (P,0.0002), suggesting senescence.

p53//p21//P63//Rb

--//Upregulation//Downregulation//--

Knockdown//SA-¦Â-gal activity assay//Western blot//RT-PCR

PC-3 cells are p53 and p16 null, RWPE-1 cells are p53 and Rb null. LNCaP cells express wild-type p53, but its levels were unchanged by DHT exposure;p21 was critical for AR-induced senescence since p21 silencing significantly reduced SA-¦ÂGal-positive population after DHT-treatment cells;p21 knock-down increased p63 protein and mRNA.AR activation also caused p63 exclusion from the nuclei,suggesting that nuclear DNp63 is important for blocking senescence. .P63 deficit likely contributed to the AR-driven senescence, since p63 knock-down elevated senescence in parental PC-3 cells.In PC3-AR and LNCaP treated with DHT, Rb phosphorylation visibly diminished.

--

Human

L

cellular senescence

22,403,609

Gene

0

1

0

1

0

PINK1

65018

protein coding

HBEC

Lung

Aging

Prevent

SA--gal activity assay//Western blot

PINK1 knockdown also enhanced CSE-induced mitochondrial ROS production and HBEC senescence.

--

Human

L

cellular senescence

25,714,760

Gene

0

1

0

1

0

PRKN

5071

protein coding

HBEC

Lung

Aging

Prevent

Knockdown//SA--gal activity assay

PARK2 knockdown enhanced HBEC senescence in response to CSE exposure as measured by SA-¦Â-gal staining and expression of CDKN1A.

--

Human

L

cellular senescence

25,714,760

Gene

0

1

0

1

0

HAS2

3037

protein coding

Fibroblast

Lung

Lung fibrosis

Prevent

BrdU assay//Flow cytometry//Knockdown//SA--gal activity assay

Deletion of HAS2 markedly suppressed the proliferation of fibrotic fibroblasts. Flow cytometry analysis revealed that HAS2 depletion significantly increased the proportion of cells in G1 phase.there was less BrdU incorporation in HAS2 siRNA transfected fibroblasts. Most interestingly, the proportion of cells with positive SA-¦Â-gal staining was dramatically increased following HAS2 depletion.

--

p27-CDK2-SKP2

--

Western blot//Knockdown

p27 protein levels were significantly increased in HAS2 deficient fibrotic fibroblasts at various time points after HAS2 siRNA transfection . Characterization of p27 in the presence of cycloheximide revealed that down-regulation of HAS2 increased p27 protein stability .CDK4 and cyclin B levels were also decreased in HAS2 deficient fibrotic fibroblasts.We observed a decrease (~ 30%) in SKP2 expression in HAS2 deficient fibrotic fibroblasts compared with control transfectants. We found that CDK2 protein levels were dramatically down-regulated upon HAS2 knock down in fibrotic fibroblasts ,which supported the hypothesis that HAS2 deletion-induced proliferative inhibition was through p27 accumulation and its effector pathway.

Human

L

cellular senescence

26,987,798

Gene

0

1

0

1

0

1

0

1

0

PLD2

5338

protein coding

IMR-90

--

Aging

Prevent

Knockdown//SA--gal activity assay

We downregulated PLD2 using siRNA in proliferating IMR-90 cells (PDL 32) and found an increased rate of SA-¦Â-gal staining in proliferating cells, compared with the control cells.

CK2¦Á

--

SA-¦Â-gal activity assay//Knockdown

Proliferating IMR-90 (PDL 32) and HCT116 cells were transfected with PLD2 siRNA, and a significant increase in the SA-¦Â-gal activity was found in response to PLD2 knockdown, whereas co-transfection of cells with CK2¦Á resulted in a significant decrease in the level of SA-¦Â-gal staining.

p53-p21

--

Knockdown//SA-¦Â-gal activity assay//Western blot

In the wild-type HCT116 cells, significant increase in SA-¦Â-gal activity in response to PLD2 knockdown was observed. However, p53- and p21Cip1/WAF1-negative cells did not show senescence£»When the protein levels of p53 and p21Cip1/WAF1were determined by Western blot, the expression of p53 was found to be upregu-lated in both the wild type and p21Cip1/WAF1-/- cells treated with PLD2 siRNA, but not in p53-/-cells. The expression of p21Cip1/WAF1 was upregulated only in the wild-type HCT116 cells treated with PLD2 siRNA.

Human

L

cellular senescence

25,064,843

Gene

0

1

0

1

0

TP53

7157

protein coding

IMR-90

--

Aging

Accelerate

SA--gal activity assay//Western blot

In TP53-deficient primary and tumour cell lines senescence decreased markedly and malic enzyme depletion lost its ability to induce this phenotype.By contrast, malic enzyme depletion did not cause cell death; it induced the expression of p53 target genes implicated in senescence but not apoptosis.

ME1//ME2

--//--

Western blot

We knocked down TP53 in human osteosarcoma U2OS cells and normal diploid fibroblast IMR90 cells using short hairpin RNA (shRNA)£¬This led to a significant increase in messenger RNA levels of ME1 and ME2, accompanied by increased protein levels and total enzymatic activity of ME1 and ME2 .

--

Human

L

cellular senescence

23,334,421

Gene

0

1

0

1

0

WT1

7490

protein coding

NCI-H522,NCI-H1437,NCI-H1568,NCI-H1650,NCI-H1975,NCI-H2126,NCI-H23,NCI-H358,NCI-H441,NCI-H460,NCI-H727,NCI-H1299,NCI-H2009,NCI-A549

--

Aging

Prevent

MTT assay//SA--gal activity assay

A significant decrease in the cell viability of cell lines harboring KRAS mutations was observed in response to WT1 loss.Mutant KRAS NSCLC cell lines adopted a senescence-like appearance in response to WT1 knockdown, and this was confirmed by staining with SA-¦Âgal.

--

Kras

--

SA-¦Â-gal activity assay//Tumor formation assay

KrasG12D/+;Wt1¦¤/¦¤ MEFs demonstrated a flattened morphology and stained positive for senescence-associated ¦Â-gal (SA-¦Âgal). Wt1 suppression significantly decreased the size of tumors when compared with controls and to a level similar to that caused by suppression of Kras.

Human

HL

cellular senescence

20,972,333

Gene

0

1

0

1

0

WFDC1

58189

protein coding

HT-1080,WI-38,IMR-90

--

Aging

Prevent

Colony formation assay//qRT-PCR

We found that overexpression of WFDC1 exerted an inhibitory effect on the colony formation efficiency of HT1080 cells£»RNAwas produced from pre-senescent and senescent cells and the expression level of WFDC1 were measured by QRT-PCR. WFDC1 mRNA level was increased as well.

--

Human

HL

cellular senescence

18,842,679

Gene

0

1

0

1

0

ATF6

22926

protein coding

HDF

--

Aging

Accelerate

Knockdown//SA--gal activity assay//qRT-PCR

ATF6¦Á silencing significantly reduced the number of SA-¦Â-Gal positive-cells from about 60% in control cells to 25% in A TF6¦Á silenced NHDFs. A small decrease of SA-¦Â-Gal positive-cells was also observed upon IRE1¦Á knock-down, but not upon PERK knockdown.

--

Human

HL

cellular senescence

27,563,820

Gene

0

1

0

1

0

1

0

PLK1

5347

protein coding

MDA-MB-468

--

Breast cancer

Prevent

DAPI staining//EdU assay//SA--gal activity assay//Western blot

PLK1 inhibition also resulted in senescence induction and p16 up-regulation in addition to p21 up-regulation in MDA-MB-468 cells.

miR-200c//miR-141//BMI1//PRC1

Downregulation//Downregulation//Upregulation//--

qRT-PCR//Western blot//Knockdown

Our data indicated that indeed PLK1 knockdown results in up-regulation of miR-200c and miR-141, while PLK1 overexpression resulted in down-regulation of miR200c and miR-141 £»Our results indicated that indeed PLK1 overexpression leads to up-regulation of BMI1 and PRC1 activity.

--

Human

L

cellular senescence

25,505,268

Gene

0

1

0

1

0

1

0

1

0

NR2E1

7101

protein coding

IMR-90

--

Aging

Prevent

BrdU assay//Colony formation assay//SA--gal activity assay//SAHF

NR2E1 expression resulted in an appreciable extension of replicative lifespan, as judged by cumulative population doublings and colony formation assays .A higher percentage of the NR2E1-expressing cells incorporated BrdU and a smaller percentage stained positively for SA-¦ÂGal activity, or showed evidence of senescence-associated heterochromatin foci£»The expression of NR2E1 partially prevented the growth arrest and senescence observed upon RAS induction.

CBX7

Upregulation

qRT¨CPCR

We observed a direct correlation, suggesting that enhanced levels of NR2E1 associated with glioblastoma multiformeformation result in a concomitant increase in CBX7 expression .

--

Human

HL

cellular senescence

25,328,137

Gene

0

1

0

1

0

1

0

1

0

SPRY1

10252

protein coding

--

Thyroid gland

Aging

Accelerate

SA--gal activity assay

Virtually all follicular cells from wild-type mice were strongly positive for SA-¦Â-Gal activity, whereas only a minority of cells from knockout mice was faintly stained. together with increased Ki67 labeling of knockout follicular cells and increased proliferation in vitro,suggest that these cells bypass a program of cellular senescence engaged in the normal thyroid glands.

--

NF-¦ÊB

--

Western blot

Levels of IkBa, which is responsible for retention of NFkB dimers in the cytoplasm, were increased in the knockout thyroid.IkBa degradation was not impaired, IkBa levels after LPS challenging remained higher in thyroids from knockout mice when compared with wild-type mice.

Human

L

cellular senescence

24,270,409

Gene

0

1

0

HBP1

26959

protein coding

2BS

--

Aging

Accelerate

BrdU assay//Growth curve assay//SA--gal activity assay

HBP1 played a role in growth suppression, as shown by BrdU incorporation assay and growth curve.HBP1 also induced premature senescence, as demonstrated by an increase of SA-¦Â-gal staining.

DNMT1

Downregulation

Western blot//RT-PCR//Knockdown

Exogenous HBP1 expression reduced DNMT1 protein (left) and mRNA (right) levels in both 2BS and WI-38 human fibroblasts.we used short hairpin RNA (shRNA) to knock down the HBP1 gene. An HBP1 knockdown increased DNMT1 protein and mRNA levels.

--

Human

L

cellular senescence

23,249,948

Gene

0

1

0

1

0

1

0

HRAS

3265

protein coding

REF

--

Aging

Accelerate

SA--gal activity assay

We confirmed that oncogenic H-Ras induced senescence as scored by senescence-associated ¦Â-Gal activity.

Myc

--

SA-¦Â-gal activity assay

Ras-induced senescence was blocked efficiently by Myc.

--

Human

L

cellular senescence

19,966,300

Gene

0

1

0

WNT16B

51384

protein coding

MRC-5

--

Aging

Accelerate

BrdU assay//SA--gal activity assay

When overexpressed in young MRC5 fibroblasts, WNT16B resulted in an increased percentage of cells positive for SA-¦Â-Gal, a decrease in cell proliferation, and a decrease in bromodeoxyuridine incorporation.

p21//p16

--//Upregulation

Western blot

Downregulation of WNT16B also prevented the accumulation of p21WAF1protein£»WNT16B overexpression induced moderate p16INK4AmRNA and protein expression.

PI3K-Akt

Activation

Western blot

AKT is also activated in WNT16Bexpressing MRC5 fibroblasts. Moreover, AKT activation was dependent on phosphoinositide 3-kinase (PI3K) activity and was abrogated through treatment with the specific PI3K inhibitor LY294002 .

Human

HL

cellular senescence

19,951,988

Gene

0

1

0

1

0

MCM

4171

protein coding

EPC2

--

Aging

Accelerate

Western blot

All of the examined MCM family members were found downregulated as EPC2 cells underwent senescence as documented by a reduced phosphorylation level of pRB protein£»the MCM2-related fragment was expressed weakly in subconfluent EPC2 cells, where subpopulation cells undergoes spontaneous senescence in primary culture.

--

Human

L

cellular senescence

19,001,876

Gene

0

1

0

PFDN5

5204

protein coding

MCF 10A,H1299

--

Aging

Accelerate

SA--gal activity assay//Western blot

SA-¦Â-Gal staining demonstrates that MM1 overexpression significantly increases the percentage of SA-¦Â-Gal-positive cells, which is a hallmark of cell senescence, the protein level of cellular senescence marker PAI-1 is elevated.

c-Myc//CD4//¦¤Np63¦Á//HERC3

Downregulation//Downregulation//--//--

Western blot//Knockdown//qRT-PCR

The IB analysis demonstrates that c-Myc and its downstream target CDK4 are dramatically down-regulated by MM1.The results showed that up-regulation of MM1 induced by depletion of HERC3 also occurs in MCF-10A cells. Further study revealed that knockdown of HERC3 abrogates ¦¤Np63¦Á-mediated down-regulation of MM1 protein levels.

--

Human

L

cellular senescence

29,880,857

Gene

0

1

0

1

0

TERF1

7013

protein coding

Bone marrow cell

Bone marrow,Peripheral blood

Bone marrow failure

Prevent

Knockdown//SA--gal activity assay//Telomere length assay

To directly assess induction of senescence in vivo caused by TRF1 deletion, we used SA-¦Â-gal actosidase staining directly on freshly isolated bone marrow cells.TRF1flox/floxMx1-Cre mice undergoing long-term Cre-induction/TRF1 deletion showed a dramatic telomere shortening of approximately 15 kb within 7-9 weeks of treatment in comparison with TRF1flox/floxMx1-wt mice after 13 weeks of treatment.

p53//p21

--//--

Western blot//Immunostaining

We found significantly greater p53 levels in pI-pC¨Ctreated TRF1flox/floxMx1-Cre mice compared with similarly treated TRF1flox/floxMx1-wt controls;a significant increase in p21 protein levels in the treated TRF1flox/floxMx1Cre group compared with the TRF1flox/floxMx1-wt controls.

--

Human

L

cellular senescence

22,932,806

Gene

0

1

0

1

0

1

0

CLCA2

9635

protein coding

MCF-7

--

Aging

Accelerate

SA--gal activity assay

We found an increase in positive staining for SA-¦Â-gal in cells expressing CLCA2 compared with Mock-transfected cells.

p53

--

qRT-PCR//Western blot//Northern blot

We selected CLCA2 for further analysis because CLCA2 was remarkably increased during the cellular senescence process as well as by the ectopic induction of wild-type p53 .We also confirmed the p53-mediated CLCA2 induction by quantita-tive RT-PCR, Northern, and Western blot analyses.

--

Human

HL

cellular senescence

22,431,922

Gene

0

1

0

DNMT3A

1788

protein coding

HCT116

--

Aging

Prevent

Knockdown//SA--gal activity assay

We transfected HCT116 cells with DNMT3a siRNA plasmid, and we found that the number and intensity of SA-¦Â-gal staining cells were increased at 1 lM Dox after DNMT3a was knocked down.

p21

--

Western blot//qRT-PCR

We found that after transfection with DNMT3a siRNA, p21 protein level was upregulated.Transfection of DNMT3a expression plasmid downregulated p21 protein level.

--

Human

HL

cellular senescence

20,473,858

Gene

0

1

0

1

0

LMNA

4000

protein coding

HCT116

--

Hutchinson-Gilford progeria syndrome

Accelerate

SA--gal activity assay

However, longerterm adenoviral overexpression of prelamin A for 7 days induced a significant increase in SA¦ÂGstaining.

--

Human

L

cellular senescence

20,458,013

Gene

0

1

0

ING1

3621

protein coding

Hs68

--

Aging

Accelerate

BrdU assay//SA--gal activity assay

Cells treated with the agents noted above were tested periodically for SA-¦Â-gal staining and BrdU incorporation,ING1a induced senescence by 36 hours and became maximal by 48 hours.

--

p16-Rb

Activation

Western blot

Consistent with ING1a inducing changes similar to those seen in replicative senescence, Rb and p16INK4a levels increased to similar degrees in response to ING1a expression and replicative senescence.

Human

L

cellular senescence

26,439,691

Gene

0

1

0

1

0

ALOX15B

247

protein coding

--

Prostate

Prostate cancer

Accelerate

SA--gal activity assay

There was a noticeable increase in SA-¦Âgal-positive glands in fl26 prostates compared with WT prostates, as previously observed.18 Important, there were also significantly more SA-¦Âgal-positive glands in the prostates of Myc;LOX animals compared with Hi-Myc mouse prostates.

p27//Rb//Rb1CC1

--// --// --

Western blot

Together, the results on p27 and Rb suggest that 15-LOX2-mediated tumor suppression in Hi-Myc model might involve cell senescence induction associated with molecular changes in these 2 molecules.The fact that increased Rb1cc1 expression was observed in 15-LOX2fl26 animals, which produce enzymatically Activate 15-LOX2 but not in WT or in 15-LOX2svb, which is enzymatically inActivate,18 suggests that 15-LOX2 metabolites may be mediating the Rb1cc1 induction.

--

Human

L

cellular senescence

24,732,589

Gene

0

1

0

PRKCI

5584

protein coding

MCF-7,T47D

--

Breast cancer

Prevent

Cell morphological analysis//SA--gal activity assay

PKCi depletion significantly increased the percentage of SA-¦Â-gal-positive cells in both MCF7 and T47D cells. Cells that were SA-¦Â-gal-positive showed an enlarged, flattened morphology.

p21

--

Western blot//Immunostaining//SA-¦Â-gal activity assay

As with MCF7 breast cancer cells, senescence occurred in the absence of a detectable DNA-damage response and was dependent on the presence of p21.

--

Human

HL

cellular senescence

22,120,720

Gene

0

1

0

1

0

SIRT1

23411

protein coding

ADSC

--

Aging

Prevent

SA--gal activity assay

Cells were treated with SRT1720, a specific SIRT1 activator. Consistent with expected result, SRT1720 decreased the number of SA-¦Âgalimmunopositive cells.

p53

Downregulation

Western blot

p53 protein was decreased by SIRT1 overexpression (pb0.05), suggesting that SIRT1 negatively regulated P53 protein expression.

--

Human

L

cellular senescence

29,803,744

Gene

0

1

0

MAP2K3

5606

protein coding

184A1,MCF 10A

--

Breast cancer

Accelerate

MTT assay//SA--gal activity assay

Overexpression of MAP2K3 in 184A1 and MCF10A cells resulted in pronounced senescence of cells.PD rate of MAP2K3-expressing cells was decreased already after 3 days, and was halted by day 6, while the control cells continued to proliferate. When SA-¦Â-galactosidase was measured, we observed that expression of MAP2K3 strongly increased its activity. Measurement of cell proliferation showed that expression of MAP2K3 had an inhibitory effect.

p53//p38//pRb

Upregulation//Upregulation//Upregulation

Western blot

We measured the levels of p53, p38, pRB and Erk1/2 in cells transfected with MAP2K3 expression vector. We observed that enhanced expression of MAP2K3 led to increase of p53, pRB and p38 expression, but not Erk1/2.

--

Human

HL

cellular senescence

21,137,025

Gene

0

1

0

1

0

EGLN1

54583

protein coding

HHUA

--

Endometrial cancer

Accelerate

Cell morphological analysis//SA--gal activity assay

Seven clones out of HHUA transfectants, 8 of Ishikawa and 9 of HWCA showed flattened cytoplasm, frequently accompanied with multinuclei formation analogous to replicative cell senescence. A significantly increased fraction of acid-¦Â-gal positive cells in the transected clones indicated the induction of cancer cell senescence by exogenous EGLN1 expression.

p21//hTERT//HIF-1¦Á

Upregulation//Downregulation//Downregulation

Western blot

In contrast, only the p21 CDK inhibitor was significantly upregulated in response to the EGLN1 transfection.Downregulation of hTERT was also observed in passage 5 of HHUA clone 3-2 and 122.As expected, both HHUA and Ishikawa cells silenced HIF-a expression in response to EGLN1 expression, followed by the suppressed expression of VEGF, which is a downstream transcriptional target of HIF-1a.

--

Human

L

cellular senescence

16,161,047

Gene

0

1

0

1

0

THBS1

7057

protein coding

HPAEC,HAEC

--

Aging

Accelerate

BrdU assay//Cell morphological analysis//Flow cytometry//MTT assay//PI staining//SA--gal activity assay

We found that TSP1 induced cell cycle arrest of HPAECs, characterized by increased numbers of cells in G1-G0 and decreased numbers of cells in S and G2-M; TSP1 significantly inhibited HPAEC proliferation, as detected by trypan blue exclusion,MTT assay , and BrdU incorporation; TSP1 exposure significantly increased the number of SA-¦Â-Gal¨Cpositive HPAECs and human aortic endothe-lial cells, which showed concentration dependency,and elicited an enlarged and flattened cell shape characteristic of senescence .

NOX1//p53//p21

Upregulation//Upregulation//Upregulation

Flow cytometry//qRT-PCR//Western blot//Immunostaining

FACS and mRNA analysis confirmed that TSP1 exposure significantly increased Nox1 abundance.We observed that Nox1 protein abundance was significantly lower in middle-aged TSP1?/?lungs compared to controls;We observed an increase in total p53 and p21cipin middle-aged mice.Moreover, we discovered that TSP1 exposure increased p53 abundance in HPAECs in a time-dependent fashion and increased p53 activation, as evidenced by its increased nuclear localization.

--

Human

L

cellular senescence

29,042,481

Gene

0

1

0

1

0

1

0

1

0

1

0

1

0

HIC1

3090

protein coding

PANC-1,BxPC-3,AsPC-1

--

Pancreatic cancer

Accelerate

Flow cytometry//MTT assay//SA--gal activity assay

Restoration of HIC1 function with 5-aza-dC treatment leaded to a reduction in cell proliferation, obvious cell senescence, cell cycle arrest and apoptosis, accompanied with acetylated p53 and p21WAF1 of Cip1 upregulation.Compared to untreated cells, cell proliferation of PANC-1, BXPC-3 and AsPC-1 treated with 5-aza-dC were significantly repressed . After treated with 5-aza-dC, the senescence positive rate of PANC-1, BXPC-3 and AsPC-1 cells were 43.6 ¡À 4.0, 69.9 ¡À 3.3 and 12.2 ¡À 2.2 %, all of which were significantly higher than in corresponding control group.Compared with control group, 5-aza-dC treated cells in G1/G0 phase were notably increased .

p53//p21/WAF1//SIRT1

Upregulation//Upregulation//Downregulation

Western blot//MTT assay//SA-¦Â-gal activity assay//Flow cytometry

After transfected with pCDNA3FlagHIC1, the acetylated p53 and p21WAF1/Cip1expression of PANC-1 cells significantly increased.After transfection with pCDNA3FlagHIC1, the cells demonstrated high-level expression of HIC1 protein and robustly downregulation of endogenous SIRT1 protein, which resulted in marked cell growth inhibition , cell senescence, cell cycle arrest and apoptosis.

--

Human

L

cellular senescence

22,552,606

Gene

0

1

0

1

0

1

0

IGFBP7

3490

protein coding

U87 MG,T98G

Glioma

Aging

Accelerate

SA--gal activity assay

U87MG and T98G cultures treated with IGFBP7 (10 ¦Ìg/ml) for 6 and 9 d, respectively, exhibited a larger number of senescence- associated ¦Â-galactosidase (SA-¦Â-gal) positive cells compared with control cultures.

--

Human

L

cellular senescence

21,795,858

Gene

0

1

0

CD38

952

protein coding

NPC,5-8F

--

Nasopharyngeal carcinoma

Prevent

CCK-8 assay//Colony formation assay//Flow cytometry//SA--gal activity assay

The effect of CD38 on the proliferation of 5-8F cells was examined using a CCK-8 assay. The results demonstrated that CD38 promoted the proliferation of 5-8F cells .Acolony formation assay was performed. The results indicated that the high expression of CD38 clone number is increased, compared with the control group; The results demonstrated that the number of ¦Â-galactosidase-positive 5-8F/CD38 cells was reduced, compared with 5-8F/Vector cells. Additionally, it was demonstrated that CD38 overexpression reduced cellular senescence in NPC cells.This demonstrated that CD38 promoted S phase DNA replication and promoted the proliferation of NPC cells.

CDK4//mitogen-activatedproteinkinase9//Cyclin D1

Upregulation//Upregulation//Upregulation

Western blot

Upregulation of cell proliferation-associated genes CDK4, mitogen-activated protein kinase 9 and Cyclin D1 progressed the cell cycle from the G1 phase to the S phase resulting in increased cell proliferation.

--

Human

L

cellular senescence

30,535,454

Gene

0

1

0

1

0

1

0

1

0

NOTCH1

4851

protein coding

HMVEC,HAEC

--

Atherosclerosis

Accelerate

Cell morphological analysis//SA--gal activity assay

We tested the effect of enforced activation of Notch1 signaling on HAEC proliferation and observed that Notch activation resulted in significantly retarded cell growth and induced an enlarged, flattened and dendritic senescent morphology.Notch activation resulted in EC senescence since more senescent((¦Â-gal©) cells were detectable from NICeGFP/HMVEC and NICeGFP/HAEC than GFP/HMVEC and GFP/HAEC.

--

Human

HL

cellular senescence

23,078,884

Gene

0

1

0

1

0

FGF21

26291

protein coding

MSC

Bone marrow

Aging

Prevent

Immunostaining//Knockdown//SA--gal activity assay

FGF21 siRNA treatment also markedly enhanced SA-¦Â-gal-positivity in P4 BM-MSCs.Ki-67 staining revealed that the number of Ki-67-positive cells was significantly reduced in FGF21 siRNA-treated P4 BM-MSCs compared with control BMMSCs.

p53//p21//Mfn2//p-Drp1

--//--//--//--

Western blot

We found that FGF21 siRNA treatment greatly reduced the protein level of FGF21 but upregulated the p53 and p21 protein level in P4 BM-MSCs,Silencing FGF21 with FGF21 siRNA elevated the level of Mfn2 and decreased the level of p-Drp1.

AMPK

--

Western blot//Knockdown

Western blotting analysis showed that silencing FGF21 with FGF21 siRNA reduced p-AMPK and p-Drp1 and upregulated the Mfn2 level in P4 BM-MSCs.

Human

L

cellular senescence

31,178,962

Gene

0

1

0

1

0

1

0

TERT

7015

protein coding

--

Lung

Inflammation

Prevent

Immunostaining//SA--gal activity assay

¦Â-Gal staining for cellular senescence was markedly increased in G3 TERT-null lung sections compared with control.We analyzed the levels of heterochromatin protein 1¦Ã(HP1¦Ã), a marker for senescence-associated heterochromatin foci, and found that immunoreActivate HP1¦Ã was also markedly increased, with the level of HP1¦Ãin G3 TERT-/- being more than 2.5 times that in WT control animals.

--

Human

L

cellular senescence

26,518,879

Gene

0

1

0

1

0

GPX7

2882

protein coding

OE33,FLO-1

--

Oesophageal adenocarcinoma

Accelerate

Colony formation assay//Flow cytometry//Growth curve assay//SA--gal activity assay//Western blot

Using stable ( pcDNA-GPX7) and transient reconstitution models of GPX7 in OAC cell lines, we detected a significant reduction in growth rates in GPX7-expressing cells.Our analysis demonstrated that GPX7-expressing OAC cells (OE33 and FLO-1) had a significant increase in ¦Â-galactosidase staining (p¡Ü0.01), as compared with control cells.About half of the control cells (Ad-CTRL) had proceeded tothe middle S phase after 4 h while only 32.4% ofGPX7-expressing cells (Ad-GPX7) proceeded to the early S phase.

--

Human

L

cellular senescence

23,580,780

Gene

0

1

0

1

0

1

0

1

0

1

0

HIC1

3090

protein coding

TPC-1

--

Tumor

Accelerate

CCK-8 assay//Flow cytometry//SA--gal activity assay

The number of cells in the G1 phase in the HIC1 transfected group (69.86 ¡À 1.22%) was slightly higher than in untransfected TPC-1 cells (62.36 ¡À 1.45%). The levels of ¦Â-galactosidase staining were also greater in the HIC1 overexpressing cells than the control TPC-1 cells. Cell proliferation in the HIC1 transfected group was significantly lower than in controls.

SIRT1

Downregulation

qRT-PCR//Western blot

The SIRT1 mRNA and protein expression levels were lowers in the HIC1 transfected group than in untransfected cells,showing that increased expression of HIC1 downregulated expression of the SIRT1 gene in TPC-1 cells.

--

Human

L

cellular senescence

27,793,057

Gene

0

1

0

1

0

1

0

HSF1

3297

protein coding

MCF 10A

--

Aging

Prevent

Cell morphological analysis//Growth curve assay

When we expressed HER2 in HSF1-depleted cells, a significant growth inhibition was observed, associated with a significant change in cells¡¯ appearance, including enlarged, flattened morphology and extensive vacuolization reminiscent of senescence.

--

Human

L

cellular senescence

20,622,894

Gene

0

1

0

1

0

ERBB2

2064

protein coding

MCF 10A

--

Aging

Accelerate

Cell morphological analysis//Growth curve assay//SA--gal activity assay

When we expressed HER2 in HSF1-depleted cells, a significant growth inhibition was observed, associated with a significant change in cells¡¯ appearance, including enlarged, flattened morphology and extensive vacuolization reminiscent of senescence.Expression of HER2 in shHSF1 MCF10A cells resulted in about 70% of ¦Â-gal-positive cells.

p21

Upregulation

Western blot

There was a marked induction of p21 upon expression of HER2 in shHSF1 cells.

--

Human

L

cellular senescence

20,622,894

Gene

0

1

0

1

0

1

0

AKT1

207

protein coding

BJ-T

--

Aging

Accelerate

Cell counting//Cell morphological analysis//SA--gal activity assay//SAHF

Cells expressing activated AKT isoforms and H-RASV12 showed a significant increase in SA-¦Â-GAL and cell size, indicating that AKT activation is sufficient to induce cellular senescence. The formation of senescence-associated heterochromatic foci (SAHF), a marker of RAS- and DNA damage-induced senescence;We found that AKT rapidly induced proliferative arrest.

p53

Upregulation

SA-¦Â-gal activity assay//Immunostaining//Knockdown//DAPI staining

A 45% increase in p53 synthesis was observed in the 30 min pulse period in myr-AKT-expressing cells compared with control cells.SA-¦Â-GAL positivity was significantly reduced in BJ-T-myr-AKT/p53 stable knockdown cells as compared with control BJ-T-myr-AKT cells.

--

Human

L

cellular senescence

21,909,130

Gene

0

1

0

1

0

1

0

1

0

SUPT5H

6829

protein coding

RKO,HT29

--

Colorectal cancer

Prevent

Colony formation assay//Knockdown//SA--gal activity assay

A higher number of senescent cells was observed in SUPT5H-knockdown cells (shSUPT5H) compared to that observed in wild-type cells or negative control cells (shNSC);Colony formation assays showed that the inhibition of SUPT5H expression effectively suppressed the colony formation efficiency of transfected cells in both monolayer cultures and soft agar, compared to that observed in wild-type cells and negative control cells (p < 0.05).

hTERT

--

Flow cytometry//Immunostaining

The results showed that inhibition of SPT5 expression by SUPT5H-specific shRNA attenuated hTERT promoter-driven GFP expression in RKO and HT29 cells transfected with hTERT promoter-driven GFP vector, whereas no obvious effect on CMV promoter-driven GFP expression was observed. Representative graphs of fluorescence-activated cell sorting (FACS) .These results indicated that SPT5 is essential for hTERT promoter activity and SUPT5H transcriptionally activated hTERT promoter-driven gene expression in human colon cancer cells.

--

Human

L

cellular senescence

26,418,880

Gene

0

1

0

1

0

1

0

CTNNB1

1499

protein coding

Ish-bcat#7,Ishikawa,Ish-tet#8

--

Endometrial cancer

Accelerate

Immunostaining//SA--gal activity assay

Some Ishbcat#7 cells with nuclear ¦Â-catenin expression demonstrated increase in size, and were more spread and flattened and multinucleated but not squamoid in appearance .After tetracycline treatment, there was an increase in numbers of senescence-like Ish-tet#8 cells similar to the case with Ish-bcat#7 cells. Four and five days after tetracycline treatment, SA-¦Â-gal-positive cells were significantly increased with Ish-tet#8, but not the mock clones.

Cyclin D1

Upregulation

Western blot

In addition, increased expression of cyclin D but not PML was evident in Ish-bcat#7 cells.

p53-p21

Upregulation

Western blot

In addition, increased expression of p53, and p21WAF1 but not PML was evident in Ish-bcat#7 cells.

Human

L

cellular senescence

15,111,320

Gene

0

1

0

1

0

RAPGEF4

11069

protein coding

EM1

--

Aging

Prevent

Knockdown//SA--gal activity assay//Western blot

The intensity of SA-¦Â-Gal staining significantly increased in the EPAC2 or CALR siRNA-treated EM1 cells compared with the control cells. Furthermore, knockdown of either EPAC2 or CALR in EM1 cells suppressed the level of p53 but increased the level of p21.

CALR

--

Western blot//Knockdown

To determine whether knockdown of EPAC2 downregulated expression of CALR protein, the level of CALR in EPAC2-silenced EM1 cells was examined by immunoblot analysis. EPAC2 knockdown significantly suppressed CALR expression.

--

Human

L

cellular senescence

25,378,661

Gene

0

1

0

1

0

1

0

CALR

811

protein coding

EM1

--

Aging

Prevent

Knockdown//SA--gal activity assay//Western blot

The intensity of SA-¦Â-Gal staining significantly increased in the EPAC2 or CALR siRNA-treated EM1 cells compared with the control cells. Furthermore, knockdown of either EPAC2 or CALR in EM1 cells suppressed the level of p53 but increased the level of p21.

CALR

--

Western blot//Knockdown

To determine whether knockdown of EPAC2 downregulated expression of CALR protein, the level of CALR in EPAC2-silenced EM1 cells was examined by immunoblot analysis. EPAC2 knockdown significantly suppressed CALR expression.

--

Human

L

cellular senescence

25,378,661

Gene

0

1

0

1

0

1

0

MED12

9968

protein coding

PC-9,SPCA1

--

Lung cancer

Prevent

Colony formation assay//Histological staining//MTT assay//SA--gal activity assay

The MTT assay showed that MED12 KO clones proliferated significantly slower from day 2 compared with the MED12 WT or MED12 reconstitution single clones. This phenomenon was further confirmed by a colony formation assay.We found that MED12 knockout caused multinucleation in PC9 cells and restoring MED12 expression in MED12 KO cells was sufficient to avert multinucleation. A similar phenomenon was observed in SPC-A1 cells. Next, we investigated the cell fate of multinucleated cells and found that they exhibited senescence.

--

Human

HL

cellular senescence

31,072,327

Gene

0

1

0

1

0

1

0

1

0

GADD45A

1647

protein coding

SCL-1

--

Skin cancer

Accelerate

Knockdown//SA--gal activity assay

Compared with the blank group,these nescence of tumor cells was postponed in the siGadd45a group (p< 0.05), while significantly improved in the Nutlin-3 group (p< 0.05). No significant difference showed in senescence among the blank, NC, and siGadd45a + Nutlin-3 groups (all p> 0.05). Cells transfected with siGadd45a had postponed senescence.

p53

--

Immunostaining

The result showed that in the tumor tissues, the positive expression rate of mutant p53 was significantly higher than the normal group (all p< 0.05).Compared with the blank group, the positive expression rate of mutant p53 was significantly increased in the siGadd45a group (p< 0.05). The Nutlin-3 group presented decreased yellow granules, indicating that mutant p53 protein expression was decreased (p< 0.05).

--

Human

L

cellular senescence

29,663,367

Gene

0

1

0

1

0

TERT

7015

protein coding

hCMEC/D3

--

AIDS

Prevent

SA--gal activity assay

¦Â-galactosidase was histochemically detectable in cells with silenced TERT (arrows), indicating that a partial loss of TERT expression is connected with accelerated hCMEC/D3 senescence.

--

Human

L

cellular senescence

20,139,322

Gene

0

1

0

SUV39H1

6839

protein coding

BEL-7402,SMMC-7721,MHCC97L,HuH-7

--

Hepatocellular carcinoma

Prevent

Cell proliferation assay//Colony formation assay//Knockdown//SA--gal activity assay

We showed that overexpression of SUV39H1 remarkably enhanced HCC cell clonogenicity, whereas SUV39H1 knockdown HCC cells reduced colony-forming ability;Knockdown of SUV39H1 significantly decreased cell proliferation and anchorage-independent growth of HCC cells;We observed an elevated senescence-associated lysosomal ¦Â-Gal activity in SUV39H1 knockdown cells.

--

Human

L

cellular senescence

22,991,213

Gene

0

1

0

1

0

1

0

1

0

BRCA1

672

protein coding

PT-5006

--

Breast cancer

Prevent

Knockdown//SA--gal activity assay//Western blot

The beta-galactosidase assay demonstrated a higher degree of cell senescence in both BRCA1-knockdown adipose-derived stem cells and breast adipose-derived stem cells with the BRCA1 mutation£»Western blotting also showed increased expression of p21, an established intermediate in the cell senescence pathway.

--

Human

L

cellular senescence

30,817,646

Gene

0

1

0

1

0

1

0

DUSP

17362377

protein coding

RPMI8226

--

Myeloma

Accelerate

SA--gal activity assay//Western blot

The increase of DUSP evidently elevated the expression of P53, while reduced the level of TLR4. However, the block of DUSP reversed the effect on P53 and TLR4, indicating that DUSP participated in H2O2 induced myeloma cell line RPMI8226 aging and might regulate the level of TLR4.

--

TLR4

Downregulation

Western blot

The increase of DUSP evidently reduced the level of TLR4.

Human

L

cellular senescence

30,280,787

Gene

0

1

0

1

0

SIRT6

51548

protein coding

KM-HM_(31)

--

Myeloma

Accelerate

SA--gal activity assay//Western blot

The increase of Sirtuin 6 apparently up-regulated the expression of P53,however, the block of Sirtuin 6 contributed to the opposite effect on P53 , indicating that Sirtuin 6 participated in CP-induced myeloma cell KM-HM_(31) aging.

--

Hippo

Downregulation

Western blot

The increase of Sirtuin 6 apparently reduced the level of Hippo. However, the block of Sirtuin 6 contributed to the opposite effect on Hippo, indicating that Sirtuin 6 might regulate the level of Hippo.

Human

L

cellular senescence

30,402,853

Gene

0

1

0

1

0

LMNA

4000

protein coding

MSC

Umbilical cord blood

Aging

Accelerate

MTT assay//SA--gal activity assay//Western blot

Immunoblots confirmed the over-expression of GFP-progerin and increased expression of p16INK4a, a marker of senescence in hMSCs;Induction of cellular senescence was also confirmed by the increase in SA-¦Â-gal activity;MTT activity was decreased in response to progerin over-expression, indicating decreased proliferative activity.

MCP-1//NF-?B

Upregulation//Upregulation

qRT-PCR//ELISA//Western blot//Immunostaining

Except for MCP-1, most cytokines that have been reported to increase in senescent fibroblasts were not increased in senescent hMSCs expressing progerin.The secretion of MCP-1 was also increased in the CM from progerin-expressing cells.The activation of NF-?B was also increased in response to progerin expression.

--

Human

L

cellular senescence

29,760,459

Gene

0

1

0

1

0

1

0

GATA4

2626

protein coding

MSC

Umbilical cord blood

Aging

Accelerate

Knockdown//SA--gal activity assay

The suppression of GATA4 in the progerin-expressing hMSCs significantly decreased SA-¦Â-gal activity;The decrease in GATA4 also inhibited the induction of senescence mediated by CM from progerin expressing cells.

MCP-1//NF-?B

--//--

qRT-PCR//Western blot

GATA4 downregulation suppressed MCP-1 gene expression in progerin expressing hMSCs, suggesting that GATA4 regulates the SASP of hMSCs through MCP-1;The activation of NF-?B was also increased in response to progerin expression£¬however,the depletion of GATA4 decreased this activity.

--

Human

L

cellular senescence

29,760,459

Gene

0

1

0

1

0

DUSP22

56940

protein coding

HS-1

--

Skin cancer

Accelerate

Western blot

Transfection of DUSP22 plasmid significantly elevated DUSP22 expression level in HS-1 skin cancer cells,inducing cell aging. Transfection of siRNA DUSP22 significantly depressed DUSP22 expression level in HS-1 skin cancer cells, decreasing expression levels of aging proteins P53 and P21.

--

MAPK

Activation

Western blot

Transfection of DUSP22 plasmid significantly elevated DUSP22 expression level in HS-1 skin cancer cells, activating MAPK .Transfection of siRNA DUSP22 significantly depressed DUSP22 expression level in HS-1 skin cancer cells,inhibiting MAPK signal pathway.

Human

L

cellular senescence

30,536,326

Gene

0

1

0

HDAC6

10013

protein coding

PDLSC

Periodontal Ligament

Aging

Prevent

BrdU assay//Cell morphological analysis//Colony formation assay//MTT assay//SA--gal activity assay

The results indicated that PDLSCs with HDAC6-inhibited by Rocilinostat or Tubastatin A displayed significantly lower proliferation rate compared with cells treated with DMSO as a control.Such inhibitory effect was further demonstrated using colony formation assay, as limited crystal violet staining of stable clones observed in HDAC6-inhibited cells versus control cells;Characteristic morphologies of senescence, including elevated SA-¦Â-gal activity, enlarged and flattened cell size, and accumulation of granular cytoplasmic inclusions could be observed in Rocilinostat or Tubastatin A treated cells rather than control cells;Moreover, reduced BrdU incorporation further supported the pro-senescence action of Rocilinostat and Tubastatin A .

p27

Downregulation

Western blot

We overexpressed HDAC6 and found significantly decreased p27Kip1 acetylation upon HDAC6 overexpression.Moreover, ectopic expression of HDAC6 also resulted in down-regulation of p27Kip1 proteins.

--

Human

L

cellular senescence

27,562,218

Gene

0

1

0

1

0

1

0

1

0

1

0

NAMPT

10135

protein coding

--

Aortas

Human Thoracic Aortic Aneurysm Disease

Prevent

SA--gal activity assay

However, in SMC-Nampt deficient mice there was SA-¦Â-gal activity throughout the ascending aorta, arch,and proximal descending thoracic aorta after 7 days of Ang II infusion.

p16

--

Western blot

This revealed a 6.1-fold increase in the abundance of p16-expressing cells in the media of the ascending aorta of Ang II-infused SMC-Nampt KO mice relative to Ang II-infused control mice and a 4.7-fold increase in the thoracic aorta.

--

Human

L

cellular senescence

28,356,339

Gene

0

1

0

VDR

7421

protein coding

MSC

Femoral head

Aging

Prevent

Cell proliferation assay//SA--gal activity assay

1,25D3 treatment of hMSC leads to a significant inhibition of cell proliferation after 48 h (17% reduction;p,0.01) and 72 h (27% reduction; p,0.001);In stimulated hMSC(gray bar) a significant reduction of ¦Â-galactosidase staining was observed compared to unstimulated cells (black bar) (p,0.001).

p16

Downregulation

qPCR

The gene expression of the senescence-associated genes cyclin dependent kinase inhibitor 2B (P15), cyclin-dependent kinase inhibitor 2A (P16), cyclin-dependent kinase inhibitor 1A (P21) and proteasome 26S subunit, non-ATPase, 9 (P27) in hMSC cultured with or without 1,25D3 for four passages led to a significant 0.4-fold(p,0.05) downregulation of P16 gene expression.

--

Human

L

cellular senescence

22,242,193

Gene

0

1

0

1

0

LMNA

4000

protein coding

Primary dermal Fibroblast

Skin

Aging

Accelerate

SA--gal activity assay

These analyses demonstrate that cells expressing progerin display a progressive, passage-dependent increase in the percentage of senescent cells over a time frame which spanned passages 5¨C10 compared to control cells ,which levels off after passage 10.

--

Human

L

cellular senescence

18,363,904

Gene

0

1

0

BMP4

652

protein coding

A549

--

Lung cancer

Accelerate

Cell morphological analysis//SA--gal activity assay

A change in morphology was apparent after 2 wk of treatment. The slower-growing cells were larger, flattened, and more granular than untreated cell;A fourfold increase in S-A-¦Â-gal activity was seen after 17 days of BMP4 treatment;Laddering was reduced in untreated A549 cells grown for 17 days in low-serum conditions making them essentially quiescent and dramatically reduced in cells treated with BMP4 for 17 days and expressing the senescence biomarker SA-¦Â-gal.

PERK//VEGF//Bcl2

Downregulation//Downregulation//Downregulation

Western blot

A decrease in proliferative and angiogenic phenotypic markers compared with untreated cells cultured under parallel low serum conditions, as shown by decreased p-ERK and VEGF expression, respectively.The expression of survival factor Bcl2 is lower in BMP4-treated cells, suggesting an increased potential for apoptosis in the senescent population.

--

Human

L

cellular senescence

12,959,928

Gene

0

1

0

1

0

E6

1489078

protein coding

SiHa

--

Cervical cancer

Prevent

Cell morphological analysis//Knockdown//SA--gal activity assay

The depression of E6 caused the SiHa/16AS cells to cease proliferation and adopt a flattened and enlarged appearance, suggesting that the cells efficiently underwent senescence.The colonies arising from the mock-transfected showed faint background staining, whereas almost all of the cells transfected with 16AS displayed intense blue staining indicative of SA-¦Â-gal activity and senescence.

--

p53-Rb

--

Western blot

Downregulation of HPV-16 E6 and E7 by 16AS transfection resulted in remarkably elevation of p53 expression in SiHa cells. Furthermore, the introduction of 16AS led to an obvious increase in the levels of hypophosphorylated p105Rb.

Human

L

cellular senescence

17,586,029

Gene

0

1

0

1

0

1

0

E7

1489079

protein coding

SiHa

--

Cervical cancer

Prevent

Cell morphological analysis//Knockdown//SA--gal activity assay

The depression of E6 caused the SiHa/16AS cells to cease proliferation and adopt a flattened and enlarged appearance, suggesting that the cells efficiently underwent senescence.The colonies arising from the mock-transfected showed faint background staining, whereas almost all of the cells transfected with 16AS displayed intense blue staining indicative of SA-¦Â-gal activity and senescence.

--

p53-Rb

--

Western blot

Downregulation of HPV-16 E6 and E7 by 16AS transfection resulted in remarkably elevation of p53 expression in SiHa cells. Furthermore, the introduction of 16AS led to an obvious increase in the levels of hypophosphorylated p105Rb.

Human

L

cellular senescence

17,586,029

Gene

0

1

0

1

0

1

0

PEA15

8682

protein coding

HDF

Foreskin

Aging

Accelerate

Cell proliferation assay//Immunostaining//Knockdown//SA--gal activity assay//Western blot

Nuclear translocation of pErk1/2 by siPEA-15 was accompanied with significant reductions of senescence markers in HDF old cells; over 10% decrease of SA-¦Â-gal activity, and 50% decrease of p53 and p21WAF1expressions.Furthermore, treatment of old cells with siPEA-15 significantly reduced senescence markers such as PML body formation, and expressions of 53BP1 and H3K9me2,as opposed to no response in young cells;The senescent cells transfected with siPEA-15 showed significant increase of cell proliferation in 2 days as compared with the siControl transfected cells,and the difference was more significant in 4 days after the transfection.

PERK1/2

--

Knockdown//Immunostaining

Knockdown of PEA-15 by siPEA-15 transfection significantly induced pErk1/2 translocation.

--

Human

L

cellular senescence

25,725,291

Gene

0

1

0

1

0

1

0

1

0

1

0

FGF2

2247

protein coding

B-MSC

Bone marrow

Aging

Prevent

SA--gal activity assay

¦Â-Gal-positive cells were highly observed in control cells and those treated with EGF and HGF, but senescent cells were detected only rarely in FGF-2-treated groups after 2 months of culture.

--

Human

L

cellular senescence

24,491,556

Gene

0

1

0

FGF4

2249

protein coding

B-MSC

Bone marrow

Aging

Prevent

SA--gal activity assay

¦Â-Gal-positive cells were highly observed in control cells and those treated with EGF and HGF, but senescent cells were detected only rarely in FGF-4-treated groups after 2 months of culture.

--

Human

L

cellular senescence

24,491,556

Gene

0

1

0

SETD1A

9739

protein coding

MDA-MB-231

--

Aging

Prevent

Cell proliferation assay//Knockdown//SA--gal activity assay

Remarkably, knockdown of SETD1A(SETD1A-KD) suppresses proliferation and triggers prompt (72h) and massive cellular senescence, with very large cells expressing characteristic ?-galactosidase (?-gal) activity.

SKP2

--

Western blot//qPCR//SA-¦Â-gal activity assay

We first confirmed that endogenous SKP2 mRNA and protein are indeed suppressed in MDA-MB-231 cells following SETD1A-KD:taining for ?-Galactosidase shows that induction of SKP2 partially suppressed the emergence of ?-Gal-positive senescent cells by 50% following SETD1A-KD.

--

Human

HL

cellular senescence

31,253,781

Gene

0

1

0

1

0

1

0

SKP2

6502

protein coding

MDA-MB-231,BT-549

--

Aging

Prevent

SA--gal activity assay

Staining for ¦Â-Galactosidase shows that induction of SKP2 partially suppressed the emergence of ?-Gal-positive senescent cells by 50% following SETD1A-KD.

p21//p27

Downregulation//Downregulation

Western blot

Inducible expression of SKP2 in SETD1A-KD cells(using two different shSET1A constructs) effectively reduced the expression of p27 and p21 proteins.

--

Human

HL

cellular senescence

31,253,781

Gene

0

1

0

CXCR2

3579

protein coding

MEF,IMR-90

Embryo

Aging

Accelerate

BrdU assay//Growth curve assay//Immunostaining//SA--gal activity assay

When viruses expressing CXCR2 were used to infect IMR-90 cells,both caused retarded growth culminating in premature senescence.The proportion of cells staining positively for senescence-associated ¦Â-ga-lactosidase (¦Â-Gal) activity and displaying senescence-associated heterochromatin foci was elevated in cultures transduced with CXCR1 compared with controls.The cells showed reduced incorporation of BrdU and displayed characteristic features of senescence.

--

p53//Rb

--//--

Crystal violet staining

To this end we analyzed the effects of CXCR2 in WI-38 human fibroblasts in which p53 or pRb functions were disrupted using the HPV E6 and E7 proteins, respectively. The data confirm the importance of p53 but imply that the Rb pathway is also involved in establishing the CXCR2-induced arrest in human cells.

Human

L

cellular senescence

18,555,777

Gene

0

1

0

1

0

1

0

1

0

NSUN2

54888

protein coding

2BS

--

Aging

Prevent

Knockdown//SA--gal activity assay

Knockdown of NSun2 inhibited cell growth and increased the proportions of cells displaying the protein marker senescence-associated (SA)-¦Â-galactosidase (~37.5% vs. ~91.0%), while overexpression of NSun2 promoted cell growth and reduced SA-¦Â-galactosidase activity (~33.5% vs. ~10.6%).

p27//CDK1//CDC25C

Downregulation//Upregulation//Upregulation

Western blot

Infection with the NSun2-expressing lentiviruses reduced the levels of p27 by ~70%, while infection with NSun2 shRNA-expressing lentiviruses increased the levels of p27 by ~5.1 fold.In contrast, the levels of proteins CDK1 and CDC25C increased in cells with overexpressed NSun2 (by ~2.8 fold for CDK1; by ~1.5 fold for CDC25C) but decreased in cells with silenced NSun2 (by ~70% for both CDK1 and CDC25C).

--

Human

HL

cellular senescence

26,687,548

Gene

0

1

0

1

0

CXCL1

2919

protein coding

NOF,OSCC cell

Mucosa

Oral cancer

Accelerate

SA--gal activity assay

Upon treatment with CXCL1 recombinant proteins, a significantly increased percentage of SA-¦Â-Gal-positive cells was detected in treated cells compared with non-treated NOFs (p < 0.05).

--

Human

L

cellular senescence

29,360,827

Gene

0

1

0

IL6

3569

protein coding

NOF,OSCC cell

Mucosa

Oral cancer

Accelerate

SA--gal activity assay

Upon treatment with IL-6 recombinant proteins, a significantly increased percentage of SA-¦Â-Gal-positive cells was detected in treated cells compared with non-treated NOFs (p < 0.05).

--

Human

L

cellular senescence

29,360,827

Gene

0

1

0

PIK3CA

5290

protein coding

EC

Umbilical cord

Vascular malformations

Accelerate

Cell morphological analysis//SA--gal activity assay

The expression of Activate PI3K evidently modified EC morphology by dramatically increasing average cell size;Indeed the expression of Activate PI3K increased the amount of ¦Â-galactosidase positive cells.

--

Human

HL

cellular senescence

29,352,118

Gene

0

1

0

1

0

CCN1

3491

protein coding

A549,NCI-H460,NCI-H520

--

Lung cancer

Accelerate

BrdU assay//Cell morphological analysis//Flow cytometry//SA--gal activity assay

The CCN1-treated H460 cells showed prominent growth retardation starting at passage number 8. For A549 and H520 cells,significant growth suppression was also observed starting at passage number 11 and 13,respectively£»Analysis by flow cytometry for cell cycle distributions of H460, A549, and H520 cells at passage number 11, 14, and 16, respectively, indicated that CCN1 induced cell cycle arrest in the G1 phase in all three NSCLC cell lines. The CCN1-treated cells exhibited a flattened morphology with concomitant cell enlargement, the presence of vacuole-rich cytoplasm, and high expression of perinuclear senescence-associated-¦Â-galactosidase.Addition of CCN1 reduced BrdU incorporation to ~20 % of that observed in control samples with no or mock treatment in all three cell lines, suggesting the cessation of DNA synthesis in the senescence-like cells.

--

p53-p21

Upregulation

Cell morphological analysis//SA-¦Â-gal activity assay//Knockdown

Addition of CCN1 to the p21-deficient cells failed to suppress cell growth, to induce senescence-like morphologies (6C), or to enhance SA-¦Â-Gal activities (6D). Similar results were found in the p53-knockdown cells.

Human

L

cellular senescence

23,553,737

Gene

0

1

0

1

0

1

0

1

0

DUSP4

1846

protein coding

WI-38

--

Aging

Accelerate

Cell morphological analysis//MTT assay//SA--gal activity assay

In contrast to the vigorously proliferating, spindle-like control fibroblasts, MKP2 cells exhibit the distinct flattened, enlarged morphology of senescent cells, sensitivity to contact inhibition, and loss of replicative capacity.we found that throughout their lifespan, the percentage of stained cells was always higher in the cultures expressing MKP2.

--

ERK

Downregulation

Knockdown

MKP2 expression resulted in a dramatic decrease of thelifespan of the ERK2:NLS line while it had no effect in the lifespan of the phosphatase resistant ERK2(D319N):NLS cells. Thus, we concluded that MKP2 overexpression inhibits proliferation and accelerates senescence by inhibiting nuclear ERK signaling and not by inactivating other MAPK family members.

Human

L

cellular senescence

17,145,763

Gene

0

1

0

1

0

1

0

RAC1

5879

protein coding

HBMEC

--

Aging

Prevent

SA--gal activity assay

Quantification of ¦Â-galactosidase positive HBMECs showed a significant increase in ¦Â-galactosidase staining following Rac 1 inhibition,suggesting that Rac 1 inhibition plays a critical role in senescence induction in the HBMECs.

--

Human

L

cellular senescence

31,513,781

Gene

0

1

0