miR-211对卵巢癌SKOV-3细胞上皮-间质转化功能的影响及机制
发布时间：2019-02-11 17:34
【摘要】：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(P0.05)。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(P0.05)。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(P0.05)。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。
[Abstract]:Objective to study the effect of microRNA-211 (miR-211) on epithelial-interstitial transformation of SKOV-3 cells and its related mechanism. Methods SKOV-3 ovarian cancer cell lines were transfected with miR-211 analogue (211M group) and negative control group (NCM group), and the untransfected control group was set up. The miR-211 content of each group was detected by RT-PCR assay. The expression levels of Snail, 伪-catenin (伪-Catenin) and high mobility gene group 4 (SOX4) were detected by Transwell assay and; Western blot assay. Western blot assay was used to detect the antagonistic effect of SOX4 overexpression on miR-211 inhibiting EMT, and double luciferase assay was used to detect the relationship between miR-211 and SOX4. Results the expression level of miR-211 in 211M group was (706.67 卤30.95) times higher than that in untransfected control group (P0.05), and the number of migration cells in 211M group was (12.32 卤0.77) / visual field. The number of invasive cells in 211M group was (9.22 卤0.32) / visual field, which was significantly lower than that in untransfected control group (82.25 卤1.05) / visual field (P0.05). The expression of Snail protein in 211M group was significantly lower than that in untransfected control group (62.10 卤1.77) / visual field (P0.05). In SOX4 211M group, the expression of Snail protein was significantly increased, and the expression of 伪 -Catenin protein was significantly decreased. Double luciferase assay showed that SOX4 was the downstream target gene of miR-211. Conclusion miR-211 inhibits the EMT function of ovarian cancer SKOV-3 cells by decreasing the SOX4 level of the downstream target gene and affecting the expression of EMT related proteins.
【作者单位】： 中山大学附属第一医院妇产科;
【基金】：国家自然科学基金青年科学基金项目(编号:81602723) 广东省科技发展专项(公益研究与能力建设)(编号:2016A020215214) 广东省医学科研基金资助项目(编号:A2015130)
【分类号】：R737.31
，
本文编号：2419950
[Abstract]:Objective to study the effect of microRNA-211 (miR-211) on epithelial-interstitial transformation of SKOV-3 cells and its related mechanism. Methods SKOV-3 ovarian cancer cell lines were transfected with miR-211 analogue (211M group) and negative control group (NCM group), and the untransfected control group was set up. The miR-211 content of each group was detected by RT-PCR assay. The expression levels of Snail, 伪-catenin (伪-Catenin) and high mobility gene group 4 (SOX4) were detected by Transwell assay and; Western blot assay. Western blot assay was used to detect the antagonistic effect of SOX4 overexpression on miR-211 inhibiting EMT, and double luciferase assay was used to detect the relationship between miR-211 and SOX4. Results the expression level of miR-211 in 211M group was (706.67 卤30.95) times higher than that in untransfected control group (P0.05), and the number of migration cells in 211M group was (12.32 卤0.77) / visual field. The number of invasive cells in 211M group was (9.22 卤0.32) / visual field, which was significantly lower than that in untransfected control group (82.25 卤1.05) / visual field (P0.05). The expression of Snail protein in 211M group was significantly lower than that in untransfected control group (62.10 卤1.77) / visual field (P0.05). In SOX4 211M group, the expression of Snail protein was significantly increased, and the expression of 伪 -Catenin protein was significantly decreased. Double luciferase assay showed that SOX4 was the downstream target gene of miR-211. Conclusion miR-211 inhibits the EMT function of ovarian cancer SKOV-3 cells by decreasing the SOX4 level of the downstream target gene and affecting the expression of EMT related proteins.
【作者单位】： 中山大学附属第一医院妇产科;
【基金】：国家自然科学基金青年科学基金项目(编号:81602723) 广东省科技发展专项(公益研究与能力建设)(编号:2016A020215214) 广东省医学科研基金资助项目(编号:A2015130)
【分类号】：R737.31
，
本文编号：2419950
本文链接：https://www.wllwen.com/yixuelunwen/fuchankeerkelunwen/2419950.html
最近更新
教材专著
