llinc000522调控Wnt通路影响胃癌侵袭转移的机制研究  

Study on the mechanism of llinc00052 regulating Wnt pathway affecting invasion and metastasis of gastric cancer

作　　者：余国庆 赫永金 许迪锋 许勇 Yu Guoqing;He Yongjin;Xu Difeng;Xu Yong(Department of the Second Surgery,Dajiangdong Hospital of Hangzhou,Hanghzou,Zhejiang 311225,China)

机构地区：[1]杭州市大江东医院外二科,311225

出　　处：《中国基层医药》2020年第2期174-178,共5页Chinese Journal of Primary Medicine and Pharmacy

基　　金：浙江省杭州市卫生计生科技计划一般项目(2018B024)。

摘　　要：目的探讨llinc000522调控Wnt通路影响胃癌侵袭转移的机制。方法选取胃癌细胞系中的SGC-7901细胞,通过长链非编码RNA(IncRNA)表达谱与逆转录定量聚合酶链锁反应(qRT-PCR)结合,筛选出与INC0005相关且在胃癌细胞中具有侵袭转移作用的微小RNA(miRNA);通过体外细胞转染实验,研究Ilinc00052和miRNA的表达变化;通过分子实验,研究Ilinc00052表达及其对Wnt/β-连环蛋白(β-catenin)表达的影响,探索linc00052能否通过调控miRNA影响Wnt/β-catenin信号通路,影响胃癌细胞的侵袭转移。结果通过Transwell小室试验测得未转染质粒组,穿膜细胞数为(134.10±4.29)个,llinc000522过表达转染质粒组穿膜细胞数为(169.24±6.99)个,差异有统计学意义(t=8.956,P=0.001)。划痕实验显示,llinc000522过表达转染质粒组迁移距离显著高于空载质粒转染组(r=0.907,P<0.01)。llinc000522过表达转染质粒组克隆形成率与空载质粒组相比明显提高[(92.75±6.32)%比(73.34±9.14)%,t=5.998,P<0.05]。与空白质粒转染相比,llinc000522过表达转染组Wnt1、Wnt3a、Wnt2、β-catenin的miRNA表达均显著上调(均P<0.05),而miRNA转染组对其表达并无明显作用,而当llinc000522、miRNA过表达共转染胃癌细胞后,Wnt1、Wnt3a、Wnt2、β-catenin miRNA转录水平提高[(1.82±0.11)、(1.52±0.15)、(1.42±0.21)、(1.71±0.19),P<0.05],但其表达仍低于单独llinc000522转染的基因miRNA转录水平。与空白质粒转染相比,llinc000522过表达转染组Wnt1、Wnt3a、Wnt2、β-catenin蛋白表达均显著上调(均P<0.05),而miRNA转染组对其表达并无明显作用,而当llinc000522、miRNA过表达共转染胃癌细胞后,Wnt1、Wnt3a、Wnt2、β-catenin蛋白表达也明显提高[(1.53±0.09)、(1.4±0.21)、(1.33±0.07)、(1.47±0.19),P<0.05],但其表达仍低于单独llinc000522转染的基因蛋白表达水平。结论在胃癌细胞中,llinc000522可通过调控miRNA水平,影响Wnt/β-catenin通路,进而影响胃癌的侵袭转移。Objective To explore the mechanism of llinc00052 regulating Wnt pathway affecting invasion and metastasis of gastric cancer.Methods SGC-7901 cells were selected from gastric cancer cell lines,and siRNAs related to INC0005 and had invasion and metastasis effects in gastric cancer cells were screened by binding of IncRNA expression profiles to qRT-PCR.Ilinc00052 and miRNA expression changes were studied by in vitro cell transfection experiments.Through molecular experiments,the expression of llinc00052 and the effect on Wnt/β-catenin expression were investigated to explore whether llinc00052 could affect the invasion and metastasis of gastric cancer cells by regulating miRNAs affecting Wnt/β-catenin signaling pathway.Results Transwell chamber test showed that the number of transmembrane cells in the untransfected plasmid group was(134.10±4.29),and the number of transmembrane cells in the overexpressed llinc000522 plasmid group was(169.24±6.99)(t=8.956,P=0.001).The scratch test showed that the migration distance in the llinc000522 overexpression transfection plasmid group was significantly higher than that in the no-load plasmid transfection group(r=0.907,P<0.01).The clone formation rate of llinc000522 overexpressed transfected plasmid group was significantly higher than that of the empty plasmid group[(92.75±6.32)%vs.(73.34±9.14)%](t=5.998,P<0.05).Compared with the transfection of blank plasmid,the expressions of Wnt1,Wnt3a,Wnt2 andβ-catenin mRNA in the llinc000522 overexpression transfection group were significantly up-regulated(P<0.05),while the miRNA transfection group had no significant effect on the expression.The expressions of Wnt1,Wnt3a,Wnt2,andβ-catenin mRNA were significantly increased[(1.82±0.11),(1.52±0.15),(1.42±0.21),(1.71±0.19)](P<0.05),but their expressions were still lower than those of the genes transfected with llinc000522 alone.Compared with the blank plasmid transfection,the expressions of Wnt1,Wnt3a,Wnt2 andβ-catenin protein in the llinc000522 overexpression transfection group wer

关 键 词：胃肿瘤 肿瘤侵润 肿瘤转移 WNT蛋白质类 长链非编码RNA 质粒 转染 克隆细胞 

分 类 号：R735[医药卫生—肿瘤]