南方医科大学学报

南方医科大学学报 ›› 2023, Vol. 43 ›› Issue (10): 1761-1770.doi: 10.12122/j.issn.1673-4254.2023.10.14

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LncRNA LINC00342通过靶向 miR-596 促进胃癌细胞的增殖、迁移和侵袭

辛宝山，刘 刚，张成雄，汪 兵，史良会   

  1. 皖南医学院第一附属医院弋矶山医院胃肠外科，安徽 芜湖 241001
  • 出版日期:2023-10-20 发布日期:2023-11-02

LncRNA LINC00342 regulates gastric cancer cell proliferation, migration and invasion by targeting miR-596

XIN Baoshan, LIU Gang, ZHANG Chengxiong, WANG Bing, SHI Lianghui   

  1. Department of Gastrointestinal Surgery, Affiliated Yijishan Hospital of Wannan Medical College, Wuhu 241001, China
  • Online:2023-10-20 Published:2023-11-02

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摘要： 目的 研究LINC00342在胃癌组织、细胞中的表达水平及影响胃癌细胞生物学功能的具体通路。方法 运用生物信息学技术，通过GEO数据库及相关预测软件，进行相关筛选及文献查询后，确定所研究的LncRNA及其下游所调控的miRNA；采用qRT-PCR检测LINC00342、miR-596在胃癌细胞系及人胃黏膜细胞中、胃癌组织及相应癌旁组织中的表达差异；细胞生物学功能上，过表达胃癌BGC-823细胞系中的LINC00342 ，将其分为pcDNA-LINC00342组和pcDNA组；过表达胃癌MGC-803细胞系中的miR-596，将其分为miR-596-mimic组及mimic-NC组；敲低胃癌MGC-803 细胞系中LINC00342的表达，将其分为si-LINC00342组及si-NC组、敲低胃癌BGC-823细胞系中miR-596的表达，将其分为miR-596-inhibitor组及inhibitor-NC组；分别进行 CCK-8、Edu细胞增殖实验、划痕实验、Transwell细胞侵袭实验、细胞周期实验以验证LINC00342 及 miR-596对胃癌细胞功能的影响 ；双荧光素酶标记实验验证LINC00342与miR-596的靶向调控关系。结果 通过生物信息学技术及相关软件预测分析，确定所研究LncRNA为 LINC00342，预测其下游调控的miRNA为miR-596；LINC00342在胃癌组织中表达高于癌旁组织（P<0.05）；在人胃癌细胞系中表达高于人胃黏膜细胞系（P<0.05），miR-596则相反（P均<0.05）；在胃癌细胞中，LINC00342 的过表达能促进胃癌细胞的增殖（P<0.05）、迁移（P<0.01）和侵袭（P<0.001），且减少处于G0/G1期的细胞比率（P<0.01）；LINC00342敲低后，胃癌细胞的增殖（P<0.05）、迁移（P<0.01）和侵袭（P<0.001）则受抑制，增加处于G0/G1期的细胞比率（P<0.01）；miR-596则发挥着相反的作用。LINC00342和miR-596能特异性结合（P=0.0067）。结论 在胃癌细胞中，LINC00342通过调控miR-596，促进胃癌细胞的增殖、迁移和侵袭。

关键词: 生物信息学技术, MGC-803 和 BGC-823细胞系, 长链非编码核糖核酸, LINC00342, miR-596

Abstract: Objective To investigate the expression levels of LINC00342 in gastric cancer (GC) tissues and cells and the pathways mediating its effects on biological behaviors of GC cells. Methods Bioinformatic analysis was performed to identify the lncRNAs and their downstream miRNAs involved in regulation of biological behaviors of GC cells. qRT-PCR was used to analyze the differential expression of LINC00342 and miR-596 in GC cell lines, human gastric mucosal cells, and GC and adjacent tissues. In human GC MGC-803 and MGC-823 cells, the effects of LINC00342 overexpression, miR-596 overexpression, LINC00342 knockdown, or miR-596 knockdown on cell proliferation, migration, invasion and cell cycle changes were examined using Edu assay, CCK-8 assay, wound healing assay, Transwell assay, and flow cytometry. The regulatory interaction between LINC00342 and miR-596 was investigated using a dual-luciferase reporter assay. Results Informatic analysis identified LINC00342 as the candidate lncRNA regulating biological behaviors of GC cells, with miR-596 as its downstream miRNA. LINC00342 expression levels were significantly higher while miR-596 expression levels were lower in GC tissues and cell lines than in the paired adjacent tissues and human gastric mucosal cell lines (all P<0.05). In MGC-803 and MGC-823 cells, overexpression of LINC00342 significantly enhanced cell proliferation (P<0.05), migration (P<0.01), and invasion (P<0.001) and reduced the percentage of G0/G1 phase cells (P<0.01), while knocking down LINC00342 significantly suppressed cell proliferation (P<0.05), migration (P<0.01), and invasion (P<0.001) and increased G0/G1 phase cell percentage (P<0.01). Modulation of miR-596 expression levels produced the opposite effects. Dual-luciferase reporter assay confirmed the specific binding between LINC00342 and miR-596 (P=0.0067). Conclusion In GC cells, LINC00342 regulates cell proliferation, migration, and invasion by targeting miR-596.

Key words: bioinformatics, MGC-803 and BGC-823 cell lines, long-stranded non-coding ribonucleic acid, LINC00342, miR-596