摘要
【目的】研究microRNA-320d(miR-320d)对子宫内膜癌JEC细胞上皮-间质转化功能的影响及其相关机制。【方法】JEC子宫内膜癌细胞株分别转染miR-320d mimics和negative control mimic,分别作为M320d、NCM组,并设立未转染对照control组,采用RT-PCR法检测各组细胞miR-320d含量。Transwell实验检测3组细胞迁移能力和侵袭能力。Westernblot法检测3组细胞α-Catenin和PBX3表达水平,及PBX3过表达对miR-320d抑制EMT的拮抗作用。双荧光素酶实验检测miR-320d与PBX3的关系。【结果】M320d组miR-320d的表达水平为control组的(808.25±15.58)倍(P<0.05)。M320d组迁移细胞数量29.56±0.59低于control组的94.48±1.02(P<0.05)。M320d组侵袭细胞数量7.33±0.84低于control组的86.28±3.51(P<0.05)。M320d组细胞α-Catenin、E-cadherin蛋白表达量(0.365±0.017、0.261±0.008)高于对照组(0.114±0.010、0.151±0.021),Vimentin蛋白表达量(0.105±0.009)低于对照组(0.211±0.025),PBX3蛋白表达量(0.181±0.002)低于对照组(0.311±0.007)。PBX3过表达M320d组子宫内膜癌细胞中α-Catenin、E-cadherin蛋白表达量(0.139±0.019、0.143±0.007)低于对照组(0.399±0.034、0.261±0.017),Vimentin蛋白表达量(0.234±0.008)高于对照组(0.105±0.009),差异均有统计学意义(P<0.05)。双荧光素酶检验结果显示PBX3为miR-320d的下游靶基因。【结论】miR-320d可能通过降低下游靶基因PBX3水平影响EMT相关蛋白表达,抑制子宫内膜癌JEC细胞的上皮-间质转化功能。
【Objective】To investigate the inhibitory effect and mechanism of the microRNA-320d（miR-320d）on epithelial mesenchymal transition in endometrial carcinoma JEC cells. 【Methods】JEC endometrial carcinoma cell lines were transfected with miR-320 d mimics or negative control mimic,respectively,as M320 d or NCM group. Control group was established with untreated JEC endometrial carcinoma cells. miR-320 d content in each group was detected by RT-PCR method. Transwell assay was used to detect the migration and invasion ability of the 3 groups. Western-blot assay was used to detect the expressions of α-Catenin,E-cadherin,Vimentin and PBX3 protein in 3 groups. Antagonistic effect of PBX3 overexpression on miR-320 d inhibition of EMT was detected by western blot assay. The relationship between miR-320 d and PBX3 was detected by dual luciferase assay.【Results】The expression level of miR-320 d in M320 d group was significantly up-regulated,and the expression level of miR-320 d was 808.25 ± 15.58 times higher than that of control group（P〈0.05）. The number of migrating cells in M320 d group was 29.56 ± 0.59,which was significantly lower than that of control group at 94.48 ± 1.02（P〈0.05）. The number of invasive cells in M320 d group was 7.33±0.84,which was significantly lower than that of group control 86.28 ± 3.51（P〈0.05）. Compared with control group,the expression ofα-Catenin and E-cadherin protein was significantly increased,the expression of Vimentin protein was significantly decreased,and the expression of PBX3 protein was significantly decreased. After PBX3 overexpression,the expression of α-Catenin and E-cadherin protein were significantly decreased,the expression of Vimentin protein were significantly increased. Dual luciferase assay showed that PBX3 is a downstream target gene of miR-320d（P〈0.05）.【Conclusion】miR-320 d may inhibit the expression of EMT related protein through the downstream target gene PBX3 and inhibit the epithelial mesenchymal transition function of endometrial carcinoma JEC cells.
作者
王晶
龚凤球
何科
姚书忠
牛刚
WANG Jing GONG Feng-qiu HE Ke YAO Shu-zhong NIU Gang(Department of Obstetrics and Gynecology, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou 510080, China)
出处
《中山大学学报（医学科学版）》
CAS
CSCD
北大核心
2017年第5期651-657,共7页
Journal of Sun Yat-Sen University:Medical Sciences
基金
国家自然科学青年基金(81602723)
广东省科技发展专项(公益研究与能力建设
2016A020215214)
广东省医学科研基金(A2015130)
