长链非编码RNAlinc00460在胃癌中的表达功能研究

目的 本研究的目的是探讨长链非编码RNAlinc00460在胃癌中的作用及其与胃癌患者临床病理特征和预后的关系。方法 通过癌症基因组图谱（TCGA）胃癌数据集系统评估linc00460的差异表达。然后，纳入88名接受胃癌手术的患者采用实时定量PCR验证TCGA结果。检测胃癌细胞株和正常人胃上皮细胞中linc00460的表达情况。采用基因集富集分析(gene set enrichment analysis，GSEA)方法预测linc00460的调控基因。将AGS细胞分别转染linc00460过表达质粒（pcDNA-linc00460）和阴性对照序列（pcDNA3.1），分别采用Western Blot检测相关蛋白表达和CCK-8法检测两组细胞的增殖情况。结果 我们发现并证实linc00460在胃癌组织和细胞中显著高表达（P < 0.05）；高linc00460表达与患者M分期相关；linc00460表达与胃癌患者总生存期之间存在强烈的负相关（P < 0.05）；且Cox比例风险模型的单变量和多变量分析证实其为胃癌相关独立生存预测因子（P < 0.05）。linc00460高表达样本富集了linc00460高表达样本富集到P53类介质对DNA损伤反应信号转导的调控(P=0.002)、P53类介质对信号转导的正向调节作用(P<0.001)、细胞分裂的正向调节(P<0.001)、核分裂的正向调节(P=0.002)等基因集。AGS细胞过表达质粒pcDNA-linc00460转染组中linc00460表达水平显著高于pcDNA3.1组，差异有统计学意义（P<0.05）。Western Blot检测结果表明linc00460可下调P53蛋白表达，上调Bcl-2和Cyclin B1表达。CCK-8结果表明pcDNA- linc00460组AGS细胞增殖指数均显著高于pcDNA3.1组，差异有统计学意义（P<0.05）。结论 linc00460在胃癌组织中高表达，且其高表达的患者预后较差，linc00460可能通过上调P53信号通路促进胃癌细胞生长，linc00460可能参与了胃癌的恶性进展。

Expression of linc00460 and its correlation with the prognosis of gastric cancerXiding Li1

Objective The purpose of this study was to investigate the expression of linc00460 in gastric cancer and its relationship with clinicopathological features and prognosis of gastric cancer patients. Methods Differential expression of linc00460 was systematically evaluated in the Cancer Genome Atlas (TCGA) gastric cancer database. Then, 88 patients underwent gastric cancer surgery were included to verify TCGA results by RT-qPCR. Linc00460 expression in AGS, HGC-27, MKN45, MGC-803 and normal gastric epithelial GES-1 were detected. Gene set enrichment analysis (GSEA) was used to predict the regulation genes of linc00460. AGS cells were transfected with pcDNA-linc00460 and pcDNA3.1, respectively. Western Blot was used to detect the expression of related proteins and CCK-8 method was used to detect the proliferation of cells in the two groups. Results We confirmed that linc00460 was highly expressed in gastric cancer tissue samples and cells (P < 0.05); moreover, high linc00460 expression was associated with M-stage; there was a strong negative correlation between the expression of linc00460 and the overall survival of patients with gastric cancer (P < 0.05); furthermore, univariate and multivariate analysis of the Cox proportional hazards model confirmed that linc00460 was an independent predictor of gastric cancer-related survival (P < 0.05). The linc00460 high-expression samples in TCGA GC datasets enriched in the regulation of DNA damage response signal transduction by P53 class mediator, positive regulation of signal transduction by P53 class mediator, regulation of meiotic nuclear division and Positive regulation of nuclear division gene sets. The expression level of linc00460 in the pcDNA-linc00460 transfected AGS cells was significantly higher than that of pcDNA3.1 group (P<0.05). Western Blot results showed that linc00460 up-regulated the P53 signaling pathway and promoted cell proliferation in AGS cells (P<0.05).Conclusion linc00460 is highly expressed in gastric cancer tissues, and patients with high linc00460 level result in a poor prognosis. Linc00460 may promote the growth of gastric cancer cells by down-regulating the P53 and upregulating Bcl-2 and Cyclin B1, and linc00460 may be involved in the malignant progression of gastric cancer.