miR-7159-5p介导TRIM26表达对胃癌细胞增殖和侵袭的影响

Effect of miR-7159-5p on the proliferation and invasion of gastric cancer cells by regulating the expression of TRIM26

目的观察微小RNA(miRNA)-7159-5p在胃癌组织中的表达,探讨其对胃癌细胞增殖和侵袭的影响及其分子机制。方法收集湖北六七二中西医结合骨科医院2018年3月至2019年11月手术切除的29例胃癌患者的癌组织和癌旁组织。通过实时荧光定量PCR(qRT-PCR)检测miR-7159-5p在癌组织和癌旁组织、癌细胞系和正常胃黏膜上皮细胞系中的表达。选择miR-7159-5p表达最低的细胞系,设置对照组和实验组,分别转染control和miR-7159-5p mimics。qRT-PCR检测转染细胞中miR-7159-5p的表达。淋巴细胞增殖检测(MTS)法检测转染细胞的增殖情况,Transwell小室法检测转染细胞的侵袭活力。miRNAMap数据库和双荧光素酶报告基因实验预测并验证miR-7159-5p的靶基因。qRT-PCR和Western blot检测转染细胞中目的基因的表达。结果胃癌组织中miR-7159-5p表达低于癌旁组织(P<0.01),胃癌细胞系miR-7159-5p表达低于正常胃黏膜上皮细胞(均P<0.01),miR-7159-5p表达最低的细胞系是SGC7901细胞(P<0.01)。转染后实验组SGC7901细胞中miR-7159-5p表达明显高于对照组(P<0.01)。转染后实验组SGC7901细胞增殖活力、侵袭活力明显低于对照组(均P<0.05)。miR-7159-5p的靶基因是三结构域蛋白26(TRIM26)。转染后实验组SGC7901细胞中TRIM26 mRNA表达明显低于对照组(P<0.01)。Western blot结果显示,转染后实验组TRIM26、c-Myc、cyclin D1、β-catenin蛋白表达量均低于对照组(均P<0.05)。结论胃癌组织中miR-7159-5p低表达,miR-7159-5p可通过下调TRIM26的表达,抑制SGC7901细胞的增殖和侵袭活力。

Objective To observe the expression of microRNA(miRNA)-7159-5p in gastric cancer tissues,and explore its effect on the proliferation and invasion of gastric cancer cells and its molecular mechanism.Methods The cancer tissues and adjacent tissues of 29 patients with gastric cancer who underwent surgical resection in Hubei 672 Orthopedic Hospital of Integrated Traditional Chinese and Western Medicine from March 2018 to November 2019 were collected.Fluorescence real-time quantitative polymerase chain reaction(qRT-PCR)was used to detect the expression of miR-7159-5p in gastric cancer tissues and adjacent tissues,gastric cancer cell lines and normal gastric epithelial cell lines.Select the cell line with the lowest expression of miR-7159-5p,set the control group and the experimental group,and be transfected with control and miR-7159-5p mimics respectively.qRT-PCR was used to detect the expression of miR-7159-5p in transfected cells.The lymphocyte proliferation test(MTS method)was used to detect the proliferation of transfected cells,and the Transwell chamber method was used to detect the invasion activity of transfected cells.The miRNAMap database and dual luciferase reporter gene experiment were used to predict and verify the target genes of miR-7159-5p.qRT-PCR and Western blot were used to detect the expression of target genes in transfected cells.Results The expression of miR-7159-5p in gastric cancer tissues was lower than that in adjacent tissues(P<0.01),the expression of miR-7159-5p in gastric cancer cell lines was lower than that of normal gastric epithelial cells(all P<0.01),and the expression of miR-7159-5p was the lowest in SGC7901 cells(P<0.01).After transfection,the expression of miR-7159-5p in SGC7901 cells of the experimental group was significantly higher than that in the control group(P<0.01).After transfection,the proliferation activity and the cell invasion activity of the experimental group was significantly lower than that of the control group(all P<0.01).The target gene of miR-7159-5p was tripartite motif 26(TRIM26).After transfection,the expression of TRIM26 mRNA in SGC7901 cells of the experimental group was significantly lower than that in the control group(P<0.01).Western blot showed that after transfection,the expression of TRIM26,c-Myc,cyclin D1,β-catenin protein were lower than those in the control group(all P<0.05).Conclusions The expression of miR-7159-5p is low in gastric cancer tissues,and miR-7159-5p can inhibit the proliferation and invasion of SGC7901 cells by down-regulating the expression of TRIM26.