miR-211对卵巢癌SKOV-3细胞上皮-间质转化功能的影响及机制

目的 研究microRNA-211(miR-211)对卵巢癌SKOV-3细胞上皮-间质转化(EMT)功能的影响及其相关机制.方法 SKOV-3卵巢癌细胞株分别转染miR-211模拟物(211M组)及其阴性对照模拟物(NCM组),并设立未转染对照组,采用RT-PCR法检测各组细胞miR-211含量;Transwell实验检测3组细胞迁移能力和侵袭能力;Western blot法检测3组细胞Snail、α-连环蛋白(α-Catenin)和与性别决定相关的高迁移率基因群4(SOX4)表达水平;采用Western blot法检测SOX4过表达对miR-211抑制EMT的拮抗作用;双荧光素酶实验检测miR-211与SOX4的关系.结果 211M组miR-211的表达水平明显上调,表达水平为未转染对照组的(706.67±30.95)倍(P<0.05).211M组迁移细胞数量为(12.32±0.77)个/视野,明显低于未转染对照组的(82.25±1.05)个/视野(P<0.05).211M组侵袭细胞数量为(9.22±0.32)个/视野,明显低于未转染对照组的(62.10±1.77)个/视野(P<0.05).211M组细胞Snail蛋白表达量明显降低,α-Catenin蛋白表达量明显升高,SOX4蛋白表达量明显降低.SOX4+211M组卵巢癌细胞中Snail蛋白表达量明显升高, α-Catenin蛋白表达量明显降低.双荧光素酶检验结果显示SOX4为miR-211的下游靶基因.结论 miR-211可能通过降低下游靶基因SOX4水平影响EMT相关蛋白表达,抑制卵巢癌SKOV-3细胞的EMT功能.

The effects and mechanism of MicroRNA-211 on epithelial mesenchymal transition function in ovarian cancer SKOV-3 cells

Objective To investigate the inhibitory effect and mechanism of the microRNA-211 (miR-211) on epithelial mesenchymal transition (EMT) in ovarian cancer SKOV-3 cells.Methods SKOV-3 ovarian cancer cell lines were transfected with miR-211 mimics or negative control mimic in 211M or NCM group, respectively.Control group was established with untreated SKOV-3 ovarian cancer cells.miR-211 content in each group was detected by RT-PCR method.Trans-well assay was used to detect the migration and invasion ability.Western-blot assay was used to detect the expression of Snail, α-Catenin and SOX4 proteins.Antagonistic effect of SOX4 over-expression on miR-211 inhibition of EMT was detected by western blot assay.The correlation between miR-211 and SOX4 was detected by dual luciferase assay.Results The expression of miR-211 in 211M group was significantly up-regulated, by 706.67±30.95 times higher than that in control group (P<0.05).The number of migrating cells in 211M group was 12.32±0.77, which was significantly lower than that in control group (82.25±1.05, P<0.05).The number of invasive cells in 211M group was 9.22±0.32, which was significantly lower than that of group control (62.10±1.77, P<0.05).Compared with control group, the expression of Snail and SOX4 proteins in 211M group was significantly reduced, while the expression of and α-Catenin protein was significantly increased.After SOX4 overexpression, the expression of Snail protein in 211M group was significantly increased, while the expression of α-Catenin protein was significantly reduced.Dual luciferase assay showed that SOX4 was a downstream target gene of miR-211 (P<0.05).Conclusion miR-211 may inhibit the expression of EMT related proteins targeting gene SOX4 and inhibit the EMT of ovarian cancer SKOV-3 cells.