compound on the cell migration. A migration assay on BMA 178-2 cells shows that the wound/scratch closed to 90%, 96% and 100% in 24 hours, 48 hours and 72 hours respectively, see However, the addition of Figures 3.25 and 3.26. AM exogenously with three different doses (1µM, 1nM and 1pM) was not able to stimulate or inhibit the migration rate of BMA178-2 cells when compared to the controls, see Figure 3.27. This result is similar to that of the Martínez et al. group, where they found that two different human breast cancer cell lines (T47D and MCF7) transfected with the AM gene did not alter their motility and migration rate, but it was shown to affect the human ovarian cancer cell line (ECV) transfected with the same AM construct (). AM22-52 which is known as the AM antagonist shows no effect on the migration of BMA178-2 in three different doses - 1µM, 1nM and 1pM compared to the control cells, see Figure 3.28. From these results it is plausible to conclude that the concentration of the AM secreted in the media by the BMA178-2 cells was high enough to prevent that action of AM22-52. This contradicts the result of Deng et al. (), where the AM22-52 inhibited the migration of the ovarian cancer cell line HO8910 cells and antagonized the stimulation effect of AM on migration. Conclusion It can be concluded that the BMA178-2 cells express RAMP1, RAMP2, RAMP3 and CLR at the mRNA and protein level which leads to the presence of the AM1, AM2 and CGRP receptors. This result suggests that AM may work in an autocrine manner to stimulate proliferation, migration and invasion as well as to reduce the apoptosis of the BMA178-2 cells through the AM2 receptor because the AM1 receptor, as illustrated in Chapter 1, plays an important role
compound on the cell migration. A migration assay on BMA 178-2 cells shows that the wound/scratch closed to 90%, 96% and 100% in 24 hours, 48 hours and 72 hours respectively, see Figures 3.25 and 3.26. However, the addition of AM exogenously with three different doses (1µM, 1nM and 1pM) was not able to stimulate or inhibit the migration rate of BMA178-2 cells when compared to the controls, see Figure 3.27. This result is similar to that of the Martínez et al. group, where they found that two different human breast cancer cell lines (T47D and MCF7) transfected with the AM gene did not alter their motility and migration rate, but it was shown to affect the human ovarian cancer cell line (ECV) transfected with the same AM construct (). AM22-52 which is known as the AM antagonist shows no effect on the migration of BMA178-2 in three different doses - 1µM, 1nM and 1pM compared to the control cells, see Figure 3.28. From these results it is plausible to conclude that the concentration of the AM secreted in the media by the BMA178-2 cells was high enough to prevent that action of AM22-52. This contradicts the result of Deng et al. (), where the AM22-52 inhibited the migration of the ovarian cancer cell line HO8910 cells and antagonized the stimulation effect of AM on migration. Conclusion It can be concluded that the BMA178-2 cells express RAMP1, RAMP2, RAMP3 and CLR at the mRNA and protein level which leads to the presence of the AM1, AM2 and CGRP receptors. This result suggests that AM may work in an autocrine manner to stimulate proliferation, migration and invasion as well as to reduce the apoptosis of the BMA178-2 cells through the AM2 receptor because the AM1 receptor, as illustrated in Chapter 1, plays an important role
Emad%27s%20Thesis%20final%20after%20correction%201-12-2015%20last.docx