坎地沙坦对卵巢癌Caov-3细胞迁移和侵袭能力的影响

Suppression effects of candesartan on proliferation,migration and invasion of ovarian cancer Caov-3 cells and its mechanism

目的:观察血管紧张素Ⅱ1型受体拮抗剂坎地沙坦(candesartan)对卵巢癌Caov-3细胞增殖、凋亡、迁移和侵袭的影响,及其与Ras同源基因(Ras homologgene,Rho)/Rho相关螺旋卷曲蛋白激酶(Rho-associatedcoiledcoil forming protein kinase,ROCK)通路相关蛋白表达的关系。方法:体外培养人卵巢癌Caov-3细胞。采用不同浓度(0.1、1、10和100μmol/L)的坎地沙坦处理Caov-3细胞,CCK-8法检测Caov-3细胞的存活率,FCM法检测Caov-3细胞的凋亡率。坎地沙坦(100μmol/L)处理Caov-3细胞24 h后,采用划痕愈合实验和Transwell小室侵袭实验分别检测Caov-3细胞的迁移及侵袭能力。蛋白质印迹法检测坎地沙坦(0.1、1、10和100μmol/L)处理24 h后,Caov-3细胞内RhoA、RhoC及ROCK1蛋白表达水平的改变,以及ROCK1蛋白下游底物肌球蛋白轻链磷酸酶(myosin lightchainphosphatase,MLCP)的功能亚基MYPT1(myosinephosphatae targeting subunit 1)磷酸化水平的改变。结果:坎地沙坦具有抑制Caov-3细胞增殖的作用(P <0.05),但对Caov-3细胞的凋亡无明显影响(P> 0.05)。坎地沙坦(100μmol/L)处理细胞24 h后能明显抑制细胞的迁移及侵袭能力(P值均<0.05);可显著下调Caov-3细胞中RhoA、RhoC及ROCK1蛋白的表达水平(P值均<0.05),以及MYPT1蛋白的磷酸化水平(P <0.01)。结论:坎地沙坦对人卵巢癌Caov-3细胞的增殖、迁移和侵袭能力具有抑制作用,这种抑制作用可能与其下调Rho/ROCK通路中RhoA、RhoC及ROCK1蛋白的表达及其下游MYPT1蛋白的磷酸化水平相关。

Objective： To investigated the effects of angiotensin H type 1 receptor （A/IR） antagonist candesartan on the proliferation, migration and invasion abilities of human ovarian cancer Caov-3 cells, and its relationship with Ras homolog gene （Rho）/Rho-associated coiled-coil forming protein kinase （ROCK） pathway-related proteins. Methods： After the treatment with different concentrations of candesartan （0.1, 1, 10 and 100 μmol/L）, the proliferation and apoptosis rate of Caov-3 cells were detected by CCK-8 and FCM, respectively. The migration and invasion abilities of Caov-3 cells treated with candesartan （100μmol/L） for 24 h were examined by the wound-healing assay and Transwell assay, respectively. The expression levels of RhoA, RhoC and ROCK1 proteins and the phosphorylation of myosine phosphatae targeting subunit 1 （MYPT1） [a functional subunit of Rho/ROCK pathway downstream substrate myosin light chain phosphatase （MLCP）] in Caov-3 cells treated with candesartan （0.1,1, 10 and 100 μmol/L） for 24 h were detected by Western blotting. Results： The growth of Caov-3 cells was significantly inhibited by candesartan （P 〈 0.05）, but the apoptotic rate of Caov-3 cells was no significantly changed （P〉0.05）. The invasion and migration abilities of Caov-3 cells treated with candesartan （100μmol/L） for 24 h were significantly inhibited （both P〈0.05）. Furthermore, candesartan could down-regulate the expression levels of RhoA, RhoC and ROCK1 proteins （all P 〈 0.05） as well as the phosphorylation level of MYPT1 （P〈0.01 ） in Caov-3 cells. Conclusion： Candesartan can inhibit the proliferation, migration and invasion of human ovarian cancer Caov-3 cells, which may be related to down-regulating the expressions of Rho/ROCK pathway-related proteins and the phosphorylation of downstream molecular MYPT1.