TIMP1在胃癌中的表达及作用机制的多数据库分析和实验研究

Multi database analysis and experimental study on the expression and mechanism of TIMP1 in gastric cancer

目的:探讨TIMP1在胃癌中的表达及作用机制。方法:应用Oncomine、GEPIA数据库分析TIMPs家族成员在胃癌及正常组织中的差异性表达,选择TIMP1为研究对象。GEPIA进一步分析TIMP1与胃癌病理分期相关性,Kaplan-Meier绘图仪分析TIMP1表达水平与胃癌患者生存预后的关系。应用CRISP/Cas9技术敲除TIMP1基因,CCK-8实验法检测细胞增殖情况,RT-PCR检测胃癌细胞AGS组与TIMP1基因敲除组的抗凋亡基因Bcl-2及促凋亡基因c-Myc的mRNA表达水平。应用DAVID数据库进行GO和KEGG分析。结果:Oncomine、GEPIA数据库分析发现胃癌组织中TIMP1表达水平更高,差异有统计学意义(P<0.01);TIMP1表达量与胃癌病理分期呈负相关;Kaplan-Meier绘图仪分析显示TIMP1高水平表达的胃癌患者较低水平表达胃癌患者生存预后更差。TIMP1基因敲除后,胃癌细胞增殖明显减少,抗凋亡基因Bcl-2 mRNA表达水平下降,促凋亡基因c-Myc mRNA表达水平上升。GO分析结果显示,TIMP1主要富集在细胞粘附分子结合、细胞外基质及细胞外结构组织等基因功能上;KEGG分析发现,TIMP1主要涉及粘着斑、癌症中的蛋白多糖等信号通路。结论:TIMP1在胃癌中呈高水平表达,与胃癌的病理分期、生存预后相关;敲除基因后,肿瘤细胞增殖减少,有望成为新的胃癌治疗靶点。

Objective: The differential expression of TIMPs family members in gastric cancer and normal tissues was analyzed by online database, TIMP1 was selected as the main analysis object. Relevant experiments further confirmed the role of TIMP1 in gastric cancer, providing clinical ideas for finding new biomarkers of gastric cancer. Methods: Oncomine and GEPIA databases were used to analyze the expression of TIMP1 in gastric cancer. GEPIA was used to further analyze the correlation between TIMP1 and pathological stage of gastric cancer. Kaplan-Meier Plotter was used to analyze the relationship between TIMP1 expression and survival prognosis of gastric cancer patients. Using CRISP/Cas9 technology to knock out TIMP1 gene. The cell proliferation was detected by CCK-8 assay. The m RNA expression levels of Bcl-2 and c-Myc were detected by RT-PCR.GO and KEGG analysis with David database. Results: Oncomine and GEPIA database analysis showed that the expression level of TIMP1 was higher in gastric cancer tissues, and the difference was statistically significant(P<0.01),The expression of TIMP1 was negatively correlated with the pathological stage of gastric cancer;Kaplan-Meier Plotter analysis showed that the survival prognosis of gastric cancer patients with high expression of TIMP1 was worse than that of gastric cancer patients with low expression of TIMP1;gene knockout test showed that the proliferation of gastric cancer cells was significantly decreased, and the m RNA expression level of Anti-apoptotic gene Bcl-2 was decreased, while the m RNA expression level of Pro-apoptotic gene c-Myc was increased. GO analysis showed that TIMP1 was mainly enriched in gene functions such as cell adhesion molecule binding, extracellular matrix and extracellular structure;KEGG analysis showed that TIMP1 was mainly involved in signal pathways such as adhesion plaque and proteoglycan in cancer.Conclusion: Comprehensive analysis of TIMPs family members found that TIMP1 was highly expressed in gastric cancer, which was related to the pathological stage and survival prognosis of gastric cancer, and was more significant than other family members;after gene knockout, the proliferation of tumor cells decreased, and the related proteins confirmed that the growth of tumor cells showed a downward trend, in order to become a new biomarker of gastric cancer.