miR-1271靶向MTDH基因调控子宫内膜癌细胞侵袭迁移的分子机制  

Molecular mechanism of miR-1271 targeting MTDH gene regulating invasion and migration of endometrial cancer cells

作　　者：王乾印 高文君 丁鑫[1] WANG Qianyin;GAO Wenjun;DING Xin(Department of Gynecology and Obstetrics,Qinghai Red Cross Hospital,Qinghai,Xi’ning 810000,China)

机构地区：[1]青海红十字医院妇产科,西宁市810000

出　　处：《河北医药》2021年第23期3539-3543,共5页Hebei Medical Journal

摘　　要：目的探讨miR-1271靶向调控MTDH对子宫内膜癌细胞侵袭迁移的影响及机制。方法将miR-1271 mimics转染子宫内膜癌RL95-2细胞,转染48h,qRT-PCR检测miR-1271基因表达;Transwell小室检测细胞侵袭和迁移能力;Western blotting检测E-cadherin、N-cadherin和Vimentin蛋白表达。采用双荧光素酶报告基因实验检测MTDH-WT/MUT与miR-1271 mimics共转染后的荧光素酶活性,Western blotting检测miR-1271 mimics/inhibitor转染RL95-2细胞后MTDH的蛋白表达,以此验证miR-1271和MTDH的靶向关系。将MTDH siRNA(si-MTDH组)、阴性对照siRNA(si-NC组)及si-MTDH+miR-1271inhibitor转染RL95-2细胞,Transwell小室及Western blotting分别检测细胞侵袭迁移及E-cadherin、N-cadherin和Vimentin蛋白表达。结果转染miR-1271 mimics的RL95-2细胞中miR-1271基因表达明显升高,过表达miR-1271及抑制MTDH表达均可明显降低RL95-2细胞侵袭和迁移能力,上调E-cadherin表达,下调N-cadherin和Vimentin表达(P<0.05),同时抑制miR-1271和MTDH表达可减弱抑制MTDH表达对细胞侵袭迁移及E-cadherin、N-cadherin和Vimentin表达的影响。结论miR-1271可通过下调MTDH表达抑制子宫内膜癌细胞侵袭和迁移。Objective To investigate the effects of and mechanism of miR-1271 targeting MTDH on invasion and migration of endometrial cancer cells.Methods The miR-1271 mimics were transfected into RL95-2 cells for 48 hours,and the expression levels of miR-1271 were detected by qRT-PCR,cell invasion and migration were detected by Transwell cell,and the expression levels of E-cadherin,N-cadherin and vimentin protein were detected by Western Blot.The luciferase activities of MTDH-WT/MUT co transfected with miR-1271 mimics were detected by double luciferase reporter gene assay,after miR-1271 mimics/inhibitor were transfected with RL95-2 cells,the expression levels of MTDH were detected by Western Blot to verify the targeting correlation between miR-1271 and MTDH.Moreover the MTDH siRNA(si-MTDH group),negative control siRNA(si-NC group)and si-MTDH+miR-1271inhibitor were transfected into RL95-2 cells,then Transwell chamber and Western Blot were used to detect the cell invasion and migration as well as the expression levels of E-cadherin,N-cadherin and vimentin protein,respectively.Results The expression levels of miR-1271 gene were significantly increased in RL95-2 cells infected with miR-1271 mimics,and the overexpression of miR-1271 and inhibition of MTDH expression could significantly reduce the invasion and migration abilities of RL95-2 cells,up regulate the expression levels of E-cadherin,down regulate the expression levels of N-cadherin and vimentin(P<0.05).Moreover to inhibit simultaneously the expressions of miR-1271 and MTDH could attenuate the effects of inhibition of MTDH expression on cell invasion and migration as well as expressions of E-cadherin,N-cadherin and vimentin.Conclusion The miR-1271 can inhibit the invasion and metastasis of endometrial cancer cells by down regulating the expression of MTDH.

关 键 词：子宫内膜癌 miR-1271 MTDH 侵袭 迁移 

分 类 号：R737.33[医药卫生—肿瘤]