雌二醇对卵巢透明细胞癌ES2细胞增殖的影响及机制探讨

The effects of 17β-estradiol on proliferation of ovarian clear cell carcinoma ES2 cell line and the possible mechanism

目的探讨17β-雌二醇(estradiol,E2)对卵巢透明细胞癌细胞系ES2细胞增殖的影响及可能机制。方法实验分为E2 10-7 mol/L组、E2 10-8 mol/L组、E2 10-9 mol/L组、E2 10-10 mol/L组、E2 10-11 mol/L组、溶剂对照组和空白对照组。采用MTS法检测不同浓度E 2作用不同时间(24,48,72 h)后各组细胞的增殖活性,以吸光度值(optical density,OD)表示;体外侵袭实验检测E 2作用后各组细胞的侵袭能力;蛋白质印记(Western blot)法检测各浓度E 2作用于ES2细胞后AT丰富结合域1A(AT rich interaction domain 1A,ARID1A)蛋白、p53蛋白、基质金属蛋白酶9(matrix metalloproteinase-9,MMP-9)蛋白表达水平的变化。结果E2作用24,48,72 h时,E2各浓度组细胞的增殖活性均高于溶剂对照组和空白对照组(P<0.05)。侵袭实验检测显示,E2各浓度组穿膜细胞数明显高于对照组(P<0.05)。Western blot法检测显示,ARID1A的表达随着E 2浓度的升高而逐渐降低,MMP-9和P53蛋白的表达随着E 2浓度的升高而增加。结论E2可使ARID1A蛋白表达量减少,p53、MMP-9蛋白表达量增加,E2增强卵巢透明细胞癌ES2细胞的增殖、侵袭能力,故卵巢透明细胞癌术后患者使用雌激素治疗时应更加慎重。

Objective To investigate effects of 17β-estradiol(E2)on cell proliferation of ovarian clear cell carcinoma cell line ES2,and to explore its possible mechanism.Methods The experiment was divided into E2 10-7 mol/L group,E2 10-8 mol/L group,E2 10-9 mol/L group,E2 10-10 mol/L group,E 2 10-11 mol/L group,solvent control group and blank control group.The proliferative activity of different E 2 concentrations at different time(24,48,72 h)was measured by MTS method and expressed as absorbance(optical density OD)value,the invasive capability was detected after treated with different concentrations of E 2 through transwell experiment.The expression of AT rich interaction domain 1A(ARID1A)protein,p53 and matrix metalloproteinase-9(MMP-9)protein in ES2 cells treated with different concentrations of E2 was measured by western blot.Results The proliferative activity of E2 group cells after treated with different concentrations of E2 for 24,48,72 hours was higher than solvent control group and blank control group(P<0.05).The transwell experiment showed that the number of transmembrane cells in the E2 group were significantly higer than that in the control group(P<0.05).Western blot analysis showed that with the increase of E2 concentration,the expression of ARID1A decreased gradually compared with the control group,while the expression of MMP-9 and p53 increased with the increased(P<0.05).Conclusion E2 could down-regulate the expressions of ARID1A protein,up-regulate the expressions of p53 and MMP-9 protein.E2 could enhance the ability of proliferation and invasion of ES2 cell.Patients with clear cell carcinoma of the ovary who had undergone surgery should be more cautious in the use of estrogen therapy.