长非编码RNA LINC01197调节胃癌进展的机制研究
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1.江苏大学附属金坛医院;2.南京医科大学第一附属医院

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国家自然科学基金项目(面上项目,重点项目,重大项目)


Long non-coding RNA LINC01197 regulates progression in gastric cancer
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    摘要:

    目的:探究长非编码RNA LINC01197在胃癌中的表达以及调节胃癌细胞进展及其相关分子机制。方法:在数据库分析和胃癌组织中检测LINC01197的表达。培养人胃癌细胞株(SGC-7901,BGC-823,MGC-803,AGS)和人胃粘膜上皮细胞(GES-1),实时荧光定量PCR(qRT-PCR)法检测LINC01197表达,CCK8,克隆形成实验检测胃癌细胞增殖,Transwell实验检测胃癌细胞侵袭迁移,裸鼠皮下成瘤实验检测胃癌细胞体内增殖。转录组测序技术(mRNA-seq)检测差异基因。qRT-PCR法、免疫印迹法分析SERPINA1相关表达水平。结果:LINC01197在胃癌组织和胃癌细胞株中表达上调 (P < 0.05);敲低LINC01197抑制胃癌细胞的增殖、侵袭和迁移(P < 0.05),以及抑制裸鼠肿瘤增殖水平(P < 0.01);LINC01197敲低后SERPINA1表达量降低(P < 0.05)。结论:LINC01197在胃癌组织和细胞中高表达,下调LINC01197可能通过抑制SERPINA1的表达从而抑制胃癌细胞进展。LINC01197有望成为胃癌治疗的新靶点。

    Abstract:

    Objective: To investigate the expression and role of long non-coding RNA LINC01197 in gastric cancer. Methods: The expression of LINC01197 was analyzed and detected by datasets and gastric cancer tissues and paracancerous tissues. Human gastric epithelial cell line (GES-1)and different gastric cancer cell lines (SGC-7901, BGC-823, MGC-803 and AGS)were cultured respectively and the expression of LINC01197 was assessed . Cell lines SGC-7901 and BGC-823 were transfected with short-hairpin RNA targeting LINC01197 (shLINC01197),qRT-PCR assays were conducted to assess LINC01197 expression. CCK8 assays and colony formation assays were carried out to evaluate proliferation,transwell assays was performed to evaluate the capability of invasion and migration. Nude murine with tumor subcutaneously implanted was to evaluate cell growth of LINC01197 in vivo. mRNA-seq assays were used to find the differential genes after LINC01197 knockdown. The expression of SERPINA1 was detected by qRT-PCR and western blotting assays. Results:The expression of LINC01197 in gastric cancer tissues and cells was higher than that in paracancerous tissues and GES-1 cells respectively (P < 0.05). The cell proliferation, invasion and migration were significantly suppressed after LINC01197 knockdown in vitro (P < 0.05). The growth ability was significantly inhibited after LIN01197 knockdown in vivo (P < 0.01). The loss of LINC01197 substantially decreased the expression of SERPINA (P < 0.05). Conclusion:LINC01197 was up-regulated in gastric cancer tissues compared to their paired paracancerous tissues and in gastric cancer cell lines compared to GES-1. Downregulation of lncRNA LINC01197 would inhibit cell proliferation,invasion and migration capability in gastric cancer cells through partly inhibiting the expression of SERPINA1. LINC01196 might be a promising target for therapy in gastric cancer.

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  • 收稿日期:2020-11-17
  • 最后修改日期:2021-02-23
  • 录用日期:2021-07-23
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