ROCK在卵巢癌中的表达及对卵巢癌细胞生物学特性影响的研究

Research on the Expression of ROCK in Ovarian Cancer and its Effect on Biological Characteristics of Ovarian Cancer Cells

目的分析ROCK在人卵巢癌中的表达,探究其对人卵巢癌A2780细胞增殖与凋亡等生物学特性的影响。方法选择66例人卵巢上皮性肿瘤,应用免疫组化法检测ROCK,并在不同浓度下的ROCK抑制剂Fasudil作用于A2780细胞,同时分别应用CCK-8检测对细胞增殖的影响,流式细胞术检测对细胞凋亡的影响,运用分光光度法对半胱氨酸蛋白酶-3的活性进行检测,Western blot检测p53蛋白的表达,探讨不同浓度Fasudil对增殖、凋亡及对caspase-3活性、p53蛋白表达的影响。结果免疫组化结果显示ROCK在卵巢上皮性恶性肿瘤阳性表达高于良性肿瘤,Ⅲ~Ⅳ期癌组织表达高于Ⅰ~Ⅱ期;和未加药组对比,不同浓度的Fasudil作用于A2780细胞24 h后,其增殖抑制率和凋亡率均升高,并依药物浓度增加而升高,caspase-3活性、p53蛋白表达量也随Fasudil浓度增高而增强。结论ROCK在卵巢癌中表达水平升高,对促进细胞增殖以及抑制凋亡中发挥一定的作用。

Objective to analyze Rho associatedcoiled coilforming proteinkinase expressed in human ovarian cancer in the exploration on human ovarian cancer A2780 cell proliferation and apoptosis of the influence of the biological characteristics,etc.Methods A total of 66 cases of human ovarian epithelial tumors were selected,ROCK was detected by immunohistochemical method,and ROCK inhibitor Fasudil was applied to A2780 cells at different concentrations.Meanwhile,the effect of CCK-8 on cell proliferation was detected,and the effect of flow cytometry on apoptosis was detected.The activity of cysteine proteinase-3 was detected by spectrophotometry,the expression of p53 protein was detected by Westernblot,and the effects of different concentrations of Fasudil on proliferation,apoptosis,caspase-3 activity and p53 protein expression were investigated.Results Immunohistochemical results showed that the positive expression of ROCK in ovarian epithelial malignant tumors was higher than that in benign tumors,and the expression of Rock in stageⅢ~Ⅳcancer tissues was higher than that in stageⅠ~Ⅱ.Compared with the non-treated group,the proliferation inhibition rate and apoptosis rate of A2780 cells increased after 24 h treatment with different concentrations of Fasudil,and the activity of caspase-3 and the expression of p53 protein also increased with the increase of Fasudil concentration.Conclusions The expression level of ROCK is increased in ovarian cancer,which plays a certain role in promoting cell proliferation and inhibiting apoptosis.