雌激素对上皮性卵巢癌细胞转移相关蛋白OPN调控作用的机制研究

The mechanism of estrogen regulation of metastasis related protein osteopontin(OPN) in epithelial ovarian cancer

背景与目的:卵巢上皮性癌具有高度侵袭性,雌激素在女性卵巢癌发生、转移中所起的作用以及肿瘤的转移、侵袭的机制研究日益引起国内外关注,骨桥蛋白(osteopontin,OPN)是一个重要的趋化因子,能促进细胞趋化、粘附和迁移,增加肿瘤细胞转移、侵袭能力,研究发现雌激素能通过信号通路影响OPN表达。本实验旨在研究雌激素对上皮性卵巢癌细胞转移相关蛋白OPN的调控作用以及可能的机制。方法:17-β雌二醇作用于卵巢浆液性腺癌细胞株SKOV3细胞,采用Transwell穿膜小室体外侵袭实验观察肿瘤细胞侵袭能力的变化;采用蛋白印迹法(Western blot)检测SKOV3细胞HIF-1α、OPN蛋白表达;以CoCl2促进HIF-1α过表达,以西罗莫司抑制HIF-1α表达后,采用Western blot方法分别检测雌激素作用下OPN蛋白表达的变化。结果:Transwell小室培养细胞24 h,雌激素组穿过微孔膜的SKOV3细胞数为162±8个,对照组为121±6个,雌激素组明显多于对照组(P=0.001)。采用Western blot方法检测,17β雌二醇促进SKOV3细胞OPN蛋白及HIF-1α蛋白表达(P=0.002,P=0.001)。促进HIF-1α过表达后,雌激素对SKOV3细胞OPN蛋白表达升调节作用增强(P=0.005);西罗莫司抑制HIF-1α表达后,雌激素对SKOV3细胞OPN蛋白表达升调节作用减弱(P=0.011)。结论:雌激素通过HIF-1α调控上皮性卵巢癌细胞转移相关蛋白OPN的表达。

Background and purpose： Estrogen has played an important role in the metastasis of epithelial ovarian cancer. Researches have revealed estrogen may regulate osteopontin （OPN）, a chemokine, to increase the invasion ability of cancer cells by signal pathway. This study is to assess estrogen＇s regulation on the expression of metastasis related protein OPN in epithelial ovarian cancer. Methods： Ovarian serous adenocarcinoma cell line SKOV3 was treated by 1713 estradiol. The invasion ability of tumor cells was detected by Transwell chambers. HIF-1α and OPN expression was measured by Western blot. The change of OPN expression when HIF-1α was over-expressed by CoC12 or suppressed by Sirolimus was measured by Western blot. Results： SKOV3 cells were cultured in transwell after 24 hours. The invasion of cells treated with estrogen was 162-4-8, significantly elevated （P=-0.001）, compared with vehicle. Western blot result showed that 17β estradiol increased expression of HIF-lct and OPN （P=-0.002, P=-0.001）. When SKOV3 cells were transfected by CoC12 which over-expressed HIF-1α, the effects of estrogen on OPN expression were attenuated （P=0.005）. When SKOV3 cells were transfected by Sirolimus which suppressed HIF-1α, the effects of estrogen on OPN expression were accenuated （P=0.011）. Conclusion： Estrogen can affect the expression of OPN by regulating HIF-1α.