整合素αvβ6缺失ERK2结合位点对SW480细胞基因表达的影响  

Effect on gene expression profile of SW480 cells after deleting ERK2 direct binding site in integrinαvβ6

作　　者：薛栋 李凡妮 肖宝安 孙祺 XUE Dong;LI Fan-ni;XIAO Bao-an;SUN Qi(Department of General Surgery,the First Affiliated Hospital of Xi′an,Jiaotong University,Xi′an 710061,China;Department of Talent Highland,the First Affiliated Hospital of Xi′an Jiaotong,University,Xi′an 710061,China)

机构地区：[1]西安交通大学第一附属医院普通外科,陕西西安710061 [2]西安交通大学第一附属医院人才高地科研平台,陕西西安710061

出　　处：《中国现代普通外科进展》2023年第7期505-510,共6页Chinese Journal of Current Advances in General Surgery

基　　金：国家自然科学基金资助项目(82073271,81702362)。

摘　　要：目的:研究整合素αvβ6缺失细胞外调节蛋白激酶2(ERK2)结合位点对结肠癌SW480细胞基因表达调控的影响。方法:构建稳定转染β6基因结构的结肠癌SW480细胞(SW480β6)和缺失ERK2结合位点的缺失突变型β6转染的SW480细胞(SW480β6Del.mutant),并通过Western blot实验检测整合素αvβ6、总ERK(t-ERK)及磷酸化ERK(p-ERK)表达水平,通过Transwell实验检测整合素αvβ6缺失ERK2结合位点对SW480细胞侵袭能力的影响。通过RNA测序研究整合素αvβ6缺失ERK2结合位点对SW480细胞基因表达谱的影响,并对差异表达基因(DEGs)进行GO和KEGG功能富集分析。结果:SW480β6和SW480β6 Del.mutant细胞均高表达αvβ6,两者之间的t-ERK表达水平差异无统计学意义(P>0.05),SW480β6 Del.mutant细胞p-ERK2水平较SW480β6细胞明显降低,且侵袭能力也明显下降(P<0.05)。通过RNA测序,在SW480β6Del.mutant细胞中共筛选出2249个DEGs,包括上调基因936个,下调基因1313个,DEGs显著富集在癌症通路、PI3K-AKT通路、MAPK通路、细胞与细胞因子受体互作方面。结论:整合素αvβ6通过αvβ6-ERK2直接连接提高ERK2的磷酸化水平,激活MAPK信号通路,促进结肠癌细胞侵袭。缺失整合素αvβ6-ERK2直接连接后,DEGs除富集于MAPK通路外,还富集于多条在结肠癌中具有重要作用的通路,提示在结肠癌中各种信号通路具有互补作用,多靶点药物对结直肠癌是一种有前景的治疗方式。Objective:To investigated the effect of the deletion of ERK2 binding site in integrinαvβ6 on the gene expression profile of colon cancer SW480 cells.Methods:The SW480 cells stably transfected withβ6 gene structure(SW480β6)and the SW480 cells transfected with deletion mutationβ6 without ERK2 binding site(SW480β6 Del.mutant)were established.The expression levels of integrinαvβ6,total ERK(t-ERK)and phosphorylated ERK(p-ERK)were detected by Western Blotting,and the effect of integrinαvβ6 without ERK2 binding site on the invasive ability of SW480 cells was detected by Transwell assay.RNA sequencing was used to study the effect of the deletion of ERK2 binding site in integrinαvβ6 on the gene expression profile of SW480 cells,and GO and KEGG functional enrichment analysis of differentially expressed genes(DEGs)were performed.Results:Both SW480β6 and SW480β6Del.mutant cells highly and equally expressedαvβ6,and there was no significant difference in t-ERK expression levels between them.In comparison with SW480β6 cells,p-ERK2 levels were significantly decreased in SW480β6 Del.mutant cells,and the invasive ability was also significantly reduced.By RNA sequencing,2249 DEGs were revealed in SW480β6 Del.mutant cells,including 936 up-regulated genes and 1313 down-regulated genes.DEGs were significantly enriched in cancer pathway,PI3K-AKT pathway,MAPK pathway,and cytokine-cytokine receptor interaction.Conclusion:The direct linkage between integrinαvβ6 and ERK2 increases the phosphorylation level of ERK2,which activates MAPK signaling pathway and promotes colon cancer cell invasion.After deleting ERK2 direct binding site in integrinαvβ6,DEGs were enriched in not only MAPK pathways but also multiple pathways that play an important role in colon cancer progression,suggesting that various signaling pathways have complementary effects in colon cancer,so multi-targeted agents are a promising treatment modality for colorectal cancer.

关 键 词：结肠肿瘤 整合素ΑVΒ6 细胞外蛋白激酶2 直接连接 RNA测序 

分 类 号：R735.35[医药卫生—肿瘤]