signature was established for lung cancer tumours (Fig. 1A). For further experiments, we focused on the characterisation of miR-99a (Fig. 1B). miR-99a reduces the proliferative capacity of lung cancer cell lines To assess the potential tumorigenic properties of miR-99a in cancer cells, the H1299, H1650 and H1975 lung cancer cell lines were transduced with miR-99a mimic (miR-99a) or miR-C (non-target control). mRNA levels of miR-99a expression were verified by qRT-PCR (Fig. 2A). miR-99a decrease proliferation NSCLC cell lines (Fig. 2B-). Proliferation curves of NSCLC cancer cells stably expressing miR-99a were similar to the transient expression experiments previously observed for the H1299, H1650 and H1975 (supplementary Fig. 4-, and data not shown). miR-99a suppresses tumorigenicity by inducing apoptosis and cell cycle arrest To uncover possible mechanisms of the miR-99amediatedsuppression of proliferation cell cycle arrest, apoptosis and senescence were studied with cells transiently expressing miR-99a. An increase in apoptosis was detected after expression in the three cell lines (Fig. 2E). Moreover, the presence of apoptosis was accompanied with cell cycle arrest in miR-99a expressing cells compared to control cells (Fig. 2F). H1299 and H1650 accumulated in G2 and H1975 in G1 phase of the cell cycle. Accordingly, a decrease in the number of miR-99a expressing cells was confirmed by Tripan-blue staining (supplementary Fig. 4). miR-99a expressing cells were negative for -galactosidase staining,
signature was established for lung cancer tumours (Fig. 1A). For further experiments, we focused on the characterisation of miR-99a (Fig. 1B). miR-99a reduces the proliferative capacity of lung cancer cell lines To assess the potential tumorigenic properties of miR-99a in cancer cells, the H1299, H1650 and H1975 lung cancer cell lines were transduced with miR-99a mimic (miR-99a) or miR-C (non-target control). mRNA levels of miR-99a expression were verified by qRT-PCR (Fig. 2A). miR-99a decrease proliferation NSCLC cell lines (Fig. 2B-). Proliferation curves of NSCLC cancer cells stably expressing miR-99a were similar to the transient expression experiments previously observed for the H1299, H1650 and H1975 (supplementary Fig. 4-, and data not shown). miR-99a suppresses tumorigenicity by inducing apoptosis and cell cycle arrest To uncover possible mechanisms of the miR-99amediatedsuppression of proliferation cell cycle arrest, apoptosis and senescence were studied with cells transiently expressing miR-99a. An increase in apoptosis was detected after expression in the three cell lines (Fig. 2E). Moreover, the presence of apoptosis was accompanied with cell cycle arrest in miR-99a expressing cells compared to control cells (Fig. 2F). H1299 and H1650 accumulated in G2 and H1975 in G1 phase of the cell cycle. Accordingly, a decrease in the number of miR-99a expressing cells was confirmed by Tripan-blue staining (supplementary Fig. 4). miR-99a expressing cells were negative for -galactosidase staining,
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