ROCK在卵巢癌组织中的表达及其对卵巢癌细胞增殖与凋亡的影响

Expression of ROCK in epithelial ovarian tumors and its effects on proliferation and apoptosis in ovarian carcinoma cells

目的:探讨Rho相关卷曲螺旋形成蛋白激酶(Rho-associated coiled-coil forming protein kinase,ROCK)在人卵巢癌组织中的表达及其对人卵巢癌SKOV3细胞增殖和凋亡的影响。方法:免疫组化SP法检测ROCK在80例人卵巢上皮性肿瘤中的表达。用ROCK抑制剂Fasudil处理SKOV3细胞,细胞计数试剂盒(cell counting kit-8,CCK-8)检测细胞增殖,流式细胞仪(flow cytometry,FCM)分析细胞凋亡水平,分光光度法检测半胱氨酸蛋白酶-3(cysteinyl aspartate specific proteinase-3,caspase-3)活性,Western blot检测凋亡相关蛋白p53表达,比较不同浓度Fasudil对细胞增殖、细胞凋亡、caspase-3活性以及凋亡相关蛋白表达的影响。结果:ROCK在恶性卵巢上皮性肿瘤的阳性表达明显高于良性肿瘤(χ2=20.961,P=0.000),在晚期上皮性卵巢癌组织中其表达高于早期肿瘤(χ2=16.675,P=0.000)。不同浓度Fasudil处理组SKOV3细胞24 h增殖抑制率和凋亡率均明显高于未加药组(F=402.537,P=0.000;F=124.251,P=0.000),并均随药物浓度增加而逐渐升高,且Fasudil呈浓度依赖性地增强caspase-3活性(F=32.423,P=0.000)和p53蛋白的表达(F=31.599,P=0.000)。结论:ROCK在卵巢癌组织中有较高表达,可能在卵巢癌细胞增殖和凋亡抑制中扮演一定角色。

Objective：To investigate the expression of Rho-associated coiled-coil forming protein kinase （ROCK） in epithelial ovarian tumors and its effects on proliferation and apoptosis in ovarian carcinoma cells. Methods：Expression of ROCK in 80 patients with epithelial ovarian tumors was assessed by immunohistochemical staining. After being treated with Fasudil, the inhibitor of ROCK, the proliferation and apoptosis of SKOV3 cells were tested by cell counting kit-8 （CCK-8） and flow cytometry（FCM） respectively,the activity of cysteinyl aspartate specific proteinase-3 （caspase-3）, caspase-3 was detected by spectrophotometry, and the expression of p53 was detected by Western blot. Results ：The expression of ROCK was higher in epithelial ovarian carcinoma tissues than in benign tumors （χ^2=20.961 ,P=0.000）, and higher in advanced carcinoma tissues than in early carcinoma tissues （χ^2=16.675,P=0.000）. Compared with the untreated SKOV3 cells, the inhibition ratios and apoptosis rates in Fasudil treated cells were increased in an dose dependent relationship （F=402.537, P=0.000 ; F= 124.251, P=0.000）. In addition, Fasudil significantly enhanced caspase-3 activity （F= 32.423,P=0.000） and increased the expression of p53 （F=31.599,P=0.000） in concentration-dependent manner. Conclusion：The expression of ROCK is higher in ovarian carcinoma than in benign tumor. ROCK may play a certain role in the proliferation and apoptosis of ovarian carcinoma cells.