目的探讨miR-372在卵巢癌细胞上皮间质转化(EMT)、侵袭迁徙中的作用及相关的机制。方法利用Lipofectamine2000瞬时转染miR-372 mimics于OVCAR3和A2780细胞中,并通过RT-PCR法检测在转染前后OVCAR3和A2780细胞中miR-372的表达,Transwell及细胞划痕实验检测OVCAR3和A2780细胞的侵袭迁徙能力,蛋白印迹法检测EMT相关蛋白N-钙黏蛋白和a-SMA蛋白的表达水平。结果转染miR-372 mimics后OVCAR3和A2780细胞的miR-372相对表达量显著升高(P〈0.05),细胞的侵袭迁徙能力显著降低(P〈0.05);Western Blotting结果表明转染miR-372 mimics后OVCAR3和A2780细胞的EMT相关间质蛋白N-钙黏蛋白和a-SMA蛋白的表达水平显著降低。结论miR-372过表达抑制卵巢癌细胞的体外侵袭及迁徙,并通过抑制N-钙黏蛋白和a-SMA蛋白水平的表达来抑制卵巢癌细胞的EMT过程。
Objective To investigate the role and mechanism of miR-372 in epithelial-mesenchymal transition(EMT), invasion and migration of ovarian cancer cells. Methods miR-372 mimics were transfected into OVCAR3 and A2780 cells by Lipofeetamine2000, the expression of miR-372 was detected by RT-PCR. Transwell and cell scratch test were used to detect the invasion and migration ability. The expression of EMT-related proteins N-cadherin and a-SMA protein was detected by Western blotting. Results The miR-372 expression level in OVCAR3 and A2780 cells was significantly increased after transfeetion of miR-372 mimics(P〈0,05). The invasion and migration ability of OVCAR3 and A2780 cells were significantly decreased after miR-372 mimics transfection(P〈0.05). Western blotting results showed that miR-372 overexpression indueed the expression level of EMT-related interstitial proteins N-cadherin and a-SMA proteins. Conclusion miR-372 overexpression inhibits ovarian cancer cell invasion and migration, and inhibits EMT via inhibiting the expression of N-eadherin and a-SMA protein in ovarian cancer cells.