摘要
目的分析长链非编码RNA(lncRNA)LHX5-AS1靶向微小RNA-532-5p(miR-532-5p)抑制胃癌细胞增殖和迁移的分子机制。方法GEPIA数据库分析LHX5-AS1在人胃癌组织中的表达水平,定量反转录聚合酶链反应(qRT-PCR)检测LHX5-AS1在人胃癌SGC7901、HS-746T、MGC803、BGC823细胞及人正常胃黏膜GES-1细胞中的表达水平。将SGC7901细胞瞬时转染阴性对照pcDNA质粒(对照组)和pcDNA-LHX5-AS1质粒(LHX5-AS1组),采用MTT实验、Transwell实验检测两组SGC7901细胞增殖、迁移能力的变化。采用双荧光素酶报告基因验证LHX5-AS1与miR-532-5p之间的相互作用,采用qRT-PCR检测LHX5-AS1过表达后miR-532-5p表达水平,采用蛋白质印迹法检测LHX5-AS1过表达后Wnt/β-catenin信号通路蛋白表达情况。结果与癌旁组织比较,LHX5-AS1在胃癌组织中表达水平明显下降(P<0.01);与GES-1细胞比较,人胃癌细胞中LHX5-AS1表达水平均明显降低(P<0.05),以SGC7901细胞中LHX5-AS1水平降低最明显(P<0.01)。与对照组比较,LHX5-AS1组SGC7901细胞的增殖、迁移能力均下降(P<0.05)。转染了野生型(WT)-LHX5-AS1和miR-532-5p的SGC7901细胞荧光素酶活性较转染了WT-LHX5-AS1和miR-NC的SGC7901细胞明显降低(P<0.01)。与对照组比较,LHX5-AS1组miR-532-5p表达水平明显下降(P<0.01),且Wnt/β-catenin信号通路蛋白Axin2、cyclin D1、c-Myc、β-catenin表达水平降低。结论LHX5-AS1在胃癌组织和细胞中表达下调,其可靶向调控miR-532-5p表达、抑制Wnt/β-catenin信号通路的活化,进而降低胃癌SGC7901细胞的增殖和迁移能力。
Objective To analyze the molecular mechanism of long non-coding RNA(lncRNA)LHX5-AS1 to inhibit the proliferation and migration of gastric cancer cells by targeting microRNA-532-5p(miR-532-5p).Methods The GEPIA database was used to analyze the expression level of LHX5-AS1 in gastric cancer tissues,and quantitative reverse transcription polymerase chain reaction(qRT-PCR)was used to detect the expression level of LHX5-AS1 in human gastric cancer SGC7901,HS-746T,MGC803,BGC823 cell and human normal gastric mucosal GES-1 cells.SGC7901 cells were transiently transfected with negative control pcDNA plasmid(the control group)and pcDNA-LHX5-AS1 plasmid(the LHX5-AS1 group),and MTT assay and transwell assay were used to detect the changes of proliferation and migration ability of SGC7901 cells in two groups.Dual-luciferase reporter gene was used to verify the interaction between LHX5-AS1 and miR-532-5p,qRT-PCR was used to detect the expression level of miR-532-5p after LHX5-AS1 overexpression,and Western blotting was used to detect the protein expression of Wnt/β-catenin signaling pathway after LHX5-AS1 overexpression.Results Compared with in adjacent tissue,the expression level of LHX5-AS1 in gastric cancer tissue significantly decreased(P<0.01);compared with in GES-1 cells,the expression level of LHX5-AS1 in human gastric cancer cells significantly decreased(P<0.05),and the level of LHX5-AS1 decreased most significantly in SGC7901 cells(P<0.01).Compared with in the control group,the proliferation and migration abilities of SGC7901 cells in the LHX5-AS1 group decreased(P<0.05).The luciferase activity of SGC7901 cells transfected with wild-type(WT)-LHX5-AS1 and miR-532-5p was significantly lower than that of SGC7901 cells transfected with WT-LHX5-AS1 and miR-NC(P<0.01).Compared with in the control group,the expression level of miR-532-5p in the LHX5-AS1 group significantly decreased(P<0.01),and the expression levels of Wnt/β-catenin signaling pathway proteins Axin2,cyclin D1,c-Myc andβ-catenin decreased.Conclusion The exp
作者
田希贵
首汉琼
魏钢
张军
杜超
王安银
TIAN Xigui;SHOU Hanqiong;WEI Gang;ZHANG Jun;DU Chao;WANG Anyin(Department of Gastrointestinal Surgery,Liangping District People′s Hospital,Chongqing 405200,China;Department of Gastrointestinal Surgery,the First Affiliated Hospital of Army Medical University,Chongqing 400038,China)
出处
《国际检验医学杂志》
CAS
2022年第15期1797-1801,共5页
International Journal of Laboratory Medicine
基金
国家自然科学基金项目(81702931)。
