The Effect And Mechanism Of Angiotensin ? On The Invasion And Migration Of Ovarian Cancer Caov-3 Cells

Objective: To investigate the effects of angiotensin II and angiotensin II type I receptor on proliferation,migration and invasion abilities of human ovarian cancer Caov-3 cells,and their relationships with Ras homologgene / Rho-associated coiled coil-forming protein kinase(Rho/ROCK)pathway-related proteins.Methods: 1.After treated with different concentrations of Ang II,candesartan or Y-27632 on 12 h,24 h,48 h,the proliferation rate of Caov-3 cells were detected by Cell Counting Kit-8.2.To further detect the effect of Candesartan on the proliferation of Caov-3 cells,Flow Cytometry(FCM)was performed to evaluate the effects of candesartan on Caov-3 cells apoptosis.3.Caov-3 cells were treated with Ang II,candesartan or Y-27632 for 24 h,then the migration and invasion abilities were detected The migration and invasion ability were examined by the wound-healing assays and Transwell assays.4.The expression of Rho A,Rho C,ROCK I and the phosphorylation of MYPT1 [a functional subunit of Rho/ROCK pathway downstream substrate myosin light chain phosphatase(MLCP)] were detected by Western blotting.Results: 1.The Cell Counting kit-8 assay revealed that,with the extention of the culture time,Ang II can promote the proliferation of ovarian cancer Caov-3 cells(P<0.01),while Candesartan and Y-27632 significantly inhibited the growth of ovarian cancer Caov-3 cells(both P<0.01);2.The apoptosis of Caov-3 cells was almost undisturbed after treated with Candesartan(P>0.05);3.In the wound-healing assay and transwell assay,Candesartan significan-tly inhibited cellular invasion(30.9±3.18)% and migration(both P<0.01).Further-more,Candesartan can down-regulate the expression of Rho A(71.04±6.96)%,Rho C(67.96±13.23)%,ROCKI(59.06±16.30)% and the phosphorylation of MYPT1(60.55±9.55)%in Caov-3 cells(all P<0.01).4.In the wound-healing assay and transwell assay,Y-27632 significantly inhibited cellular invasion(34.4±2.43)% and migration(both P<0.01).Furthermore,Y-27632 can down-regulate the expression of Rho A(69.30±7.46)%(P<0.01),Rho C(68.78±11.17)%(P<0.01),ROCKI(78.06±6.41)%(P<0.05)and the phosphorylation of MYPT1(56.36±7.74)%(P<0.01)in Caov-3 cells.5.The wound-healing assays and Transwell assays showed that exogenous Ang II could promote the invasion and migration ability of Caov-3 cell(all P<0.05),up-regulate the protein expression of Rho A(112.87±1.50)%,ROCK I(121.20±6.66)% and the phosphorylation of MYPT1(116.32±3.35)% in Caov-3 cells(all P<0.01).However,all of these effects were inhibited by Candesartan and Y-27632.Conclusion: 1.Inhibition of Rho/ROCK pathway may down-regulate the proliferation,migration and invasion abilities of human ovarian cancer Caov-3 cells.2.Antagonistic AT1 R may cause a decrease in the proliferation,migration and invasion abilities of human ovarian cancer Caov-3 cells by inhibiting the Rho/ROCK pathway,but it has no obvious effect on the apoptosis of the cells.3.Exogenous Ang II binds to AT1 R which promotes the invasion of ovarian cancer Caov-3 cells by upregulated the Rho A/ROCK pathway.