沉默RORα通过活化Wnt/β-catenin通路靶基因促进人胃癌细胞增殖  

Silencing RORαpromote the proliferation of human gastric cancer cells by activating the Wnt/β-catenin pathway

作　　者：裴大兵 张翼臻 刘芳 苏坚 夏红 苏琦 PEI Da-Bing;ZHANG Yi-Zhen;LIU Fang(Cancer Research Institute,Hengyang Medical College,Hunan Province Key Laboratory of Cancer Cellular and Molecular Pathology,Center for Gastric Cancer Research of Hunan Province,Hengyang 421001,Hunan,China)

机构地区：[1]南华大学衡阳医学院肿瘤研究所、湖南省肿瘤细胞与分子病理学重点实验室、湖南省胃癌研究中心,湖南衡阳421001 [2]井冈山大学附属医院普外科 [3]南华大学衡阳医学院附属第二医院病理科

出　　处：《中国老年学杂志》2023年第6期1408-1413,共6页Chinese Journal of Gerontology

基　　金：国家自然科学基金(No.81374013);湖南省卫计委课题(No.B2015-182)。

摘　　要：目的探讨人胃癌细胞中维甲酸相关孤核受体(ROR)α与Wnt/β-连环素(catenin)通路的相互作用。方法噻唑蓝(MTT)实验验证沉默RORα对胃癌细胞系MGC803的增殖作用;实时定量聚合酶链反应(RT-PCR)、Western印迹与免疫共沉淀检测敲低RORα对Wnt/β-catenin通路相关分子与靶基因表达;荧光素酶报告检测c-Myc基因启动子活性。结果敲低RORα可显著促进胃癌细胞增殖(P<0.05)。当RORα被敲低时可显著上调Wnt1 mRNA和蛋白表达(P<0.05);但对β-catenin无明显影响(P>0.05)。免疫共沉淀提示敲低RORα可显著降低RORα与β-catenin的结合效率(P<0.05)。RORα敲低后,核内β-catenin与转录因子(TCF)-4表达水平明显升高(P<0.05)。Wnt通路下游基因Axin、c-Myc、c-Jun和基质金属蛋白酶(MMP)-9的mRNA和蛋白水平显著上调(P<0.05)。荧光素酶报告提示,敲低RORα可显著增强c-Myc启动子活性(P<0.05)。结论敲低RORα可活化Wnt/β-catenin信号通路从而促进胃癌细胞增殖。Objective To investigate the interaction between retinoic acid-associated solitary nucleus receptors(ROR)αand Wnt/β-catenin pathway in human gastric cancer cells.Methods Multiple talbe tourmament(MTT)assay was used to verify the effect of knockdown RORαon the proliferation of gastric cancer cell line MGC803.Real-time quantitative polymerase chain reaction(RT-PCR),Western blot and immunoprecipitation were used to verify the knockdown of RORαexpression on Wnt/β-catenin pathway related molecules and target genes.The luciferase report was used to detect the promoter activity of c-Myc gene.Results Knocking down RORαcould significantly inhibit the proliferation of gastric cancer cells(P<0.05).When RORαwas knocked down,Wnt1 mRNA and protein expression were significantly up-regulated(P<0.05).But there was no significant effect onβ-catenin(P>0.05).Immunoprecipitation indicated that knocking down RORαcould significantly reduce the binding efficiency of RORαtoβ-catenin(P<0.05).After RORαknockdown,the expression levels ofβ-catenin in the nucleus and transcription factor(TCF)-4 were significantly increased(P<0.05).mRNA and protein levels of Axin,c-Myc,c-Jun and matrix metalloproteinase(MMP)-9 downstream genes of Wnt pathway were significantly up-regulated(P<0.05).The luciferase report suggested that the knockdown of RORαcould significantly enhance the activity of c-Myc promoter(P<0.05).Conclusions Knocking down;RORαcould activate the Wnt/β-catenin signaling pathway and promote the proliferation of gastric cancer cells.

关 键 词：维甲酸相关孤核受体(ROR)α 沉默 人胃癌MGC803细胞 增殖 WNT/Β-CATENIN通路 靶基因 

分 类 号：R735[医药卫生—肿瘤] R329.24[医药卫生—临床医学]