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目的研究雌激素对体外培养的卵巢透明细胞癌细胞系ES2增殖、侵袭力的影响和对MTA3、Snail、基质金属蛋白酶2(MMP2)表达的调节作用,以探讨雌激素作用对ES2细胞系的影响。方法体外培养卵巢透明细胞癌ES2细胞系,去激素培养7d后加入17β雌二醇(E2)1×10-7、1×10-8、1×10-9mol/L,MTT法检测细胞增殖情况,流式细胞仪测定细胞周期,改良的Boyden小室检测细胞侵袭力、运动力的变化,RTPCR检测MTA3、Snail、MMP2mRNA表达,免疫组化法检测Snail、MMP2表达,明胶酶谱检测基质金属蛋白酶活性。结果RTPCR、WesternBlot结果示ES2细胞系为ERα阴性表达,ERβ阳性表达,Ecad为阴性表达。17βE21×10-7、1×10-8、1×10-9mol/L均可促进细胞增殖;E21×10-8mol/L可使G0～G1期细胞减少,S期细胞增加,但对细胞凋亡无影响;17βE21×10-8mol/L可显著增强ES2细胞的侵袭力和趋向运动能力,并可降低MTA3mRNA表达,增加Snail、MMP2mRNA及蛋白表达。结论17βE2可显著促进ES2细胞的侵袭能力,此作用可能是通过17βE2对MTA3表达下调和对Snail、MMP2表达的上调实现的。
Objective To investigate the effect of estrogen on the proliferation and invasion of ovarian clear cell carcinoma ES2 cell line and the regulation of MTA3, Snail and MMP2 expression in esophageal carcinoma cell line ES2 to investigate the effect of estrogen on ES2 cell line . Methods The ovarian clear cell carcinoma ES2 cell lines were cultured in vitro. Seven days after the exogenous hormones were cultured, 17β-estradiol (E2) 1 × 10-7, 1 × 10-8 and 1 × 10-9 mol / L were added respectively. The cell proliferation was detected by MTT assay , Cell cycle was measured by flow cytometry, cell invasiveness and motility were detected by modified Boyden chamber. The expressions of MTA3, Snail and MMP2 mRNA were detected by RTPCR. The expressions of Snail and MMP2 were detected by immunohistochemistry. The activity of MMPs . Results The results of RTPCR and Western Blot showed that ES2 cell line was negative for ERα, positive for ERβ, and negative for Ecad. 17βE21 × 10-7, 1 × 10-8, 1 × 10-9mol / L all can promote cell proliferation; E21 × 10-8mol / L can G0 ~ G1 phase cells, S phase cells increased, but the cell apoptosis 17βE21 × 10-8mol / L can significantly enhance the invasiveness of ES2 cells and tend to exercise capacity, and can reduce the MTA3mRNA expression, increase Snail, MMP2mRNA and protein expression. Conclusion 17βE2 can significantly promote the invasiveness of ES2 cells. This effect may be through 17βE2 down-regulation of MTA3 and up-regulation of Snail and MMP2 expression.