MiR-610通过下调双微体基因2的表达抑制卵巢癌细胞的侵袭和迁移*

目的：探讨miR-610 对卵巢癌细胞侵袭和迁移能力的作用及机制。方法：通过RT-PCR 检测卵巢癌组织及细 胞、正常组织及细胞中miR-610 与鼠双微体基因2（MDM2）mRNA的表达；通过Lipofectamine 2000 将miR-NC、 miR-610 mimic、miR-6101 inhibitor、pc-MDM2质粒分别或联合转染进入HO8910细胞；划痕实验检测细胞迁移 能力，Transwell 法检测细胞侵袭能力，免疫印迹检测E- 钙黏着蛋白（E-cadherin）、N- 钙黏着蛋白（N-cadherin） 和MDM2蛋白表达水平，双荧光素酶报告基因检测miR-610 与MDM2靶向关系。 结果：与正常卵巢组织和上皮 细胞系HOSEpiC相比，人宫颈癌组织和细胞系中miR-610 表达明显下调，选择下调效果较明显的HO8910细胞系 进行后续实验。与Control 组相比，miR-610 mimic 组卵巢癌HO8910细胞划痕闭合率明显降低、侵袭细胞数目明 显减少、E-cadherin 明显上调、N-cadherin 表达明显下调，miR-610 inhibitor 组卵巢癌HO8910细胞划痕闭合率明显 升高、侵袭细胞数目明显增多、E-cadherin 明显下调、N-cadherin 表达明显上调。MiR-610 与MDM2 3'UTR 区存在 结合位点，且miR-610 靶向下调MDM2 表达。共转染pc-MDM2逆转了miR-610 mimic 对卵巢癌HO8910细胞迁移 和侵袭能力抑制作用。 结论：MiR-610 通过靶向下调MDM2抑制卵巢癌HO8910细胞迁移和侵袭能力。

MiR-610 inhibits invasion and migration of ovarian cancer cells by downregulating double minute 2 expression

Objective To explore the effect and mechanism of miR-610 on the invasion and migration ability of ovarian cancer cells. Methods The expression of miR-610 and murine double minute 2（MDM2）mRNA in tissues and cells from ovarian cancer and nomal condition was detected by RT-PCR. The plasmids miR-NC， miR- 610 mimic， miR-6101 inhibitor and pc-MDM2 were transfected into HO8910 cells separately or in combination by Lipofectamine 2000. The cell migration ability was tested by scratch test. The cell invasion ability was detected by Transwell method. The expression of E-cadherin， N-cadherin and MDM2 protein was detected by Western blotting. The dual luciferase report was used to detect the targeting relationship between miR-610 and MDM2. Results Compared with normal ovarian tissue and epithelial cell line HOSEpiC， miR-610 expression was significantly downregulated in human cervical cancer tissues and cell lines， and HO8910 cell line with a more significant effect of down-regulation was selected for subsequent experiments. Compared with the control group， the scratch closure rate of ovarian cancer HO8910 cells in the miR-610 mimic group was significantly reduced， the number of invading cells was significantly reduced， E-cadherin was significantly up-regulated， and N-cadherin expression was significantly down-regulated. Compared with the control group， the scratch closure rate of ovarian cancer HO8910 cells in the miR-610 inhibitor group was significantly increased， the number of invading cells was significantly increased， E-cadherin was significantly down-regulated， and N-cadherin expression was significantly up-regulated. MiR-610 had a binding site with MDM2 3'UTR region， and miR-610 targeted down-regulation of MDM2 expression. Cotransfection of pc-MDM2 reversed miR-610 mimic's inhibitory effect on ovarian cancer HO8910 cell migration and invasion. Conclusion MiR-610 inhibits the migration and invasion of ovarian cancer HO8910 cells by targeting down-regulation of MDM2.