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目的:探讨在卵巢癌细胞株A2780中,雌激素通过与不同受体结合对miR-214产生的调控作用。方法:通过在卵巢癌细胞株A2780中转染包裹雌激素受体α和β的脂质体,建立稳定表达受体的A2780-α与A2780-β。雌激素组:添加10-6mol/L的17-β雌二醇处理A-2780、A2780-α、A2780-β,于0、6、12和24 h分别提取3组细胞的RNA,采用RT-PCR法检测miR-214的表达。拮抗剂组:先在A-2780、A2780-α、A2780-β3组细胞株中加入100 n M的17-β雌二醇受体拮抗剂(ICI182、780)处理6 h,然后再添加17-β雌二醇处理12 h,分别提取3组细胞的RNA,采用RT-PCR法检测miR-214的表达。结果:10-6mol/L 17β雌二醇作用于各细胞株后,A-2780、A2780-α、A2780-β的miR-214表达量分别为0.802±0.367、0.094±0.09和1.325±0.678。miR-214表达在A2780-α较A-2780降低,A2780-β较A-2780升高。以上结果能够被17-β雌二醇受体拮抗剂ICI182、780抑制。结论:雌激素能够通过与ERα结合而降低miR-214的表达,而与ERβ结合则结果相反,表明雌激素能通过与不同的雌激素受体结合对细胞内基因进行差异性调控,这也许能为卵巢癌发病的机制及治疗提供新的思路。
AIM: To investigate the regulatory effect of estrogen on miR-214 production in ovarian cancer cell line A2780 by binding to different receptors. Methods: A2780-α and A2780-β were stably expressed by transfection of liposomes encapsulating estrogen receptor α and β in ovarian cancer cell line A2780. Estrogen: A-2780, A2780-α and A2780-β were treated with 10-6mol / L 17-β-estradiol. The RNA of three groups were extracted at 0, 6, 12 and 24 h respectively. PCR method to detect the expression of miR-214. Antagonist Group: 100 nM 17-beta estradiol receptor antagonist (ICI 182, 780) was added to A-2780, A2780-α and A2780-β3 cell lines for 6 h and then 17- β estradiol for 12 h, RNA of three groups were extracted, and the expression of miR-214 was detected by RT-PCR. Results: After treated with 10-6mol / L 17β-estradiol, the expression of miR-214 in A-2780, A2780-α and A2780-β were 0.802 ± 0.367, 0.094 ± 0.09 and 1.325 ± 0.678, respectively. miR-214 expression was lower in A2780-α than A-2780 and A2780-β was higher than A-2780. The above result can be inhibited by the 17-beta estradiol receptor antagonist ICI182,780. Conclusion: Estrogen can reduce the expression of miR-214 by binding to ERα, while the opposite result with ERβ indicates that estrogen can differentially regulate intracellular genes through binding to different estrogen receptors, which may be able to Provide a new way for the pathogenesis and treatment of ovarian cancer.