长链非编码RNA LINC01197调节胃癌进展的机制研究

Long non-coding RNA LINC01197 regulates progression in gastric cancer

目的:探究长链非编码RNA LINC01197在胃癌中的表达以及调节胃癌细胞进展的相关分子机制。方法:基于GEPIA数据库分析胃癌组织中LINC01197的表达。培养人胃黏膜上皮细胞(GES-1)和人胃癌细胞株(SGC-7901、BGC-823、MGC-803、AGS),实时荧光定量PCR(qPCR)法检测LINC01197表达,采用短发夹RNA转染SGC-7901和BGC-823细胞干扰LINC01197的表达,CCK-8、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。qPCR、免疫印迹法分析SERPINA1相对表达水平。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),并且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增殖、侵袭和迁移。

Objective:To investigate the expression and role of long non-coding RNA LINC01197 in gastric cancer. Methods:The expression of LINC01197 in gastric cancer tissues and paracancerous tissues was analyzed based on GEPIA database. 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 examined by real-time fluorescent quantitative PCR(qPCR). Cell lines SGC-7901 and BGC-823 were transfected with short-hairpin RNA targeting LINC01197(shLINC01197),CCK-8 assays and colony formation assays were carried out to evaluate cell proliferation,transwell assays was performed to evaluate the capability of invasion and migration. Nude mice were implanted with SGC-7901 and BGC-823 subcutaneously to evaluate tumor cell growth in vivo. mRNA-seq assay was used to screen the differential expressed genes after LINC01197 knockdown. The expression of SERPINA1 was detected by qPCR and Western blot. Results:The expressions of LINC01197 in gastric cancer tissues and cell lines were higher than those in paracancerous tissues and GES-1 cells respectively(P < 0.05). The cell proliferation,invasion and migration were significantly suppressed after LINC01197 knockdown in gastric cancer cell lines(P < 0.05). The tumor growth in vivo was significantly inhibited after LIN01197 knockdown(P < 0.01). The knockdown 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. Down-regulation of LINC01197 would inhibit cell proliferation,invasion and migration capability in gastric cancer cells through partly inhibiting the expression of SERPINA1.