基于生物信息学分析胃癌中RUNX1表达及其临床意义

Expression and clinical significance of RUNX1 in gastric cancer based on bioinformatics

目的研究Runt相关转录因子1(Runt-related transcription factor 1,RUNX1)在胃癌中的表达及其与胃癌患者临床病理特征、预后和肿瘤细胞侵袭能力之间的相关性。方法①数据库分析:应用TCGA和GEO数据库分析RUNX1在胃癌和癌旁组织中的表达差异;利用Kaplan-Meier Plotter数据库分析RUNX1表达水平与胃癌患者生存预后的相关性;利用GO分析和KEGG通路富集分析RUNX1在胃癌中可能参与的功能和信号通路,并通过GEPIA数据库进行基因相关性验证。②临床病例验证:回顾性收集兰州大学第二医院在2018年6月至2019年12月期间收治的62例胃癌患者的癌组织和癌旁组织,进行免疫组织化学染色、HE染色和天狼星红染色,并探索RUNX1表达与患者临床病理特征和预后的关系。③细胞实验:利用小干扰RNA(si-001-RUNX1和si-002-RUNX1)进行RUNX1的敲减,并通过Transwell实验分析RUNX1与AGS和HGC27细胞侵袭能力的关系。结果①数据库分析:RUNX1在胃癌组织中高表达,并与胃癌患者的总生存时间(overall survival,OS)呈负相关(P<0.001);GO分析和KEGG通路富集分析结果提示,RUNX1不仅参与细胞外基质(extracellular matrix,ECM)中胶原的构建,而且在ECM-受体相互作用通路中显著富集;GEPIA数据库基因相关性分析结果显示,RUNX1与ECM-受体相互作用通路所涉及基因表达呈正相关(P<0.05)。②临床病例验证:免疫组织化学染色结果提示,RUNX1在胃癌组织中相对高表达,且RUNX1高表达是影响胃癌患者术后OS的危险因素(RR=5.074,P=0.034);且胃癌组织中RUNX1表达与天狼星红红色面积呈正相关关系(r=0.46,P<0.001)。③细胞实验:Transwell实验证实RUNX1敲减后,si-001-RUNX1和si-002-RUNX1组的AGS和HGC27侵袭细胞数量降低。结论RUNX1在胃癌中高表达且提示更差的生存预后,并且RUNX1促进胃癌发生发展有可能是通过激活ECM-受体相互作用通路实现的。

Objective To investigate the expression of Runt-related transcription factor 1(RUNX1)in gastric cancer and its correlation with clinicopathological features,prognosis and tumor cell invasion ability.Methods①Database analysis:the expression of RUNX1 in gastric cancer and adjacent tissues were analyzed by TCGA and GEO database.Kaplan-Meier Plotter database was used to analyze the correlation between RUNX1 expression level and overall survival(OS)of gastric cancer patients.GO analysis and KEGG pathway enrichment were used to analyze the possible functions and signaling pathways of RUNX1 in gastric cancer,and gene correlation was verified by GEPIA database.②Clinical case validation:the cancer tissues and adjacent tissues of 62 patients with gastric cancer admitted to the Second Hospital of Lanzhou University from June 2018 to December 2019 were retrospectively collected for immunohistochemical staining,HE staining and Sirius red staining,and the relation between RUNX1 expression and clinicopathological features and prognosis of patients was explored.③Cell experiment:we knocked down RUNX1 by using small interfering RNA,and then analyzed the relation between RUNX1 and the invasion ability of gastric cancer cells by Transwell assay.Results ① Database analysis: RUNX1 was highly expressed in gastric cancer tissues and negatively correlated with OS (P<0.001).GO analysis and KEGG pathway enrichment analysis showed that RUNX1 was not only involved in the construction ofcollagen in extracellular matrix (ECM), but also significantly enriched in ECM-receptor interaction pathway. The resultsof GEPIA gene correlation analysis showed that RUNX1 was positively correlated with gene expression involved in ECMreceptor interaction pathway (P<0.05). ② Clinical case validation: the results of immunohistochemical staining showedthat RUNX1 was relatively highly expressed in gastric cancer tissues, and the high expression of RUNX1 was a risk factoraffecting the postoperative OS of gastric cancer patients (RR=5.074, P=0.034);the expression of RUNX1 in gastric cancertissues was positively correlated with red staining area of Sirius red staining (r=0.46, P<0.001). ③ Cell experiment:invasion experiments confirmed that the number of invasive AGS or HGC27 cells in si-001 group and si-002 groupdecreased after RUNX1 knockdown. Conclusion RUNX1 is highly expressed in gastric cancer and suggests a worsesurvival prognosis, and it is possible that RUNX1 promotes the development of gastric cancer by activating the ECMreceptor interaction pathway.