摘要
目的:研究鸟嘌呤核苷酸交换因子40(recombinant human rho guanine nucleotide exchange factor 40,ARHGEF40)对结肠癌细胞侵袭能力和上皮间质转化(epithelial-mesenchymal transition,EMT)标志蛋白表达的影响,探索ARHGEF40在结肠癌细胞侵袭过程中的作用及机制。方法:通过免疫组化方法检测ARHGEF40在结肠癌组织及正常肠上皮组织中的表达情况;选取人结肠癌细胞SW620、CL187为研究对象,采用细胞转染技术、侵袭实验(Transwell)以及免疫印迹实验(Western blot,WB)观察ARHGEF40对结肠癌细胞侵袭能力以及对EMT标志蛋白表达水平的影响;通过对TCGA数据库进行KEGG通路富集分析,推测ARHGEF40促进结肠癌EMT过程的机制。结果:免疫组化结果显示,ARHGEF40在正常肠上皮组织中表达阳性率为22.2%(6/27),在结肠癌组织中表达阳性率为77.1%(37/48),二者比较具有显著性差异(P<0.01)。WB结果显示,ARHGEF40在四种结肠癌细胞系中的表达显著高于其在正常肠上皮细胞系中的表达。Transwell实验结果表明,ARHGEF40过表达明显促进结肠癌CL187、SW620细胞的侵袭能力,而敲减ARHGEF40能够明显抑制CL187、SW620细胞的侵袭能力(P<0.05)。结肠癌CL187细胞系中ARHGEF40过表达明显上调Snail、Slug、Vimentin及N-cadherin的表达,同时抑制上皮样标志蛋白E-cadherin的表达;相反,在结肠癌SW620细胞系中敲减ARHGEF40能够抑制Snail、Slug、Vimentin及N-cadherin的表达,同时上调上皮样标志蛋白E-cadherin的表达。此外,采用KEGG信号通路富集分析表明,ARHGEF40过表达可能激活结肠癌中TGF-β、WNT、NOTCH和MAPK信号通路。结论:ARHGEF40可能通过激活相关信号通路促进结肠癌细胞EMT和侵袭。
Objective:To investigate the effect of ARHGEF40 on the invasion of colon cancer cells and the expression of epithelial-mesenchymal transition marker proteins.To explore the mechanism via which ARHGEF40 regulates the invasion of colon cancer cells.Methods:Immunohistochemistry assay was performed to detect the expression of ARHGEF40 in colon cancer tissues and normal intestinal epithelial tissues.Transwell and Western blot assays were performed to determine the effect of ARHGEF40 overexpression and knockdown on colon cancer cells invasion and EMT marker protein expression.KEGG pathway enrichment analysis on the TCGA database was used to predict the possible mechanism of ARHGEF40 affecting the invasion of colon cells and EMT process.Results:Immunohistochemical analyses showed that the positive rate of ARHGEF40 in normal intestinal epithelial tissue was 22.2%(6/27),and in colon cancer tissue was 77.1%(37/48),which had a significant difference(P<0.01).Western blot results revealed that ARHGEF40 was highly expressed in four colon cancer cell lines compared to normal intestinal epithelial cell line.Transwell assays indicated that overexpression of ARHGEF40 significantly promoted the invasion of colon cancer cells CL187 and SW620,where as knockdown of ARHGEF40 significantly inhibited the invasion of these two cells(P<0.05).We also found that ARHGEF40 overexpression increased the expression of Snail,Slug,Vimentin,N-cadherin and decreased the expression of epithelioid marker protein E-cadherin in CL187 cell line.On the contrary,ARHGEF40 suppression downregulated the expression of Snail,Slug,Vimentin,N-cadherin and upregulated the expression of E-cadherin in SW620 cell line.Furthermore,enrichment analysis of KEGG signalling pathway showed that ARHGEF40 may activate TGF-β,WNT,NOTCH and MAPK signalling pathways in colon cancer.Conclusion:ARHGEF40 may promote the EMT and invasion of colon cancer cells by activating related signalling pathways.
作者
顾坚
王艺华
于涓瀚
GU Jian;WANG Yihua;YU Juanhan(Department of Pathology,College of Basic Medical Sciences,China Medical University,Liaoning Shenyang 110001,China)
出处
《现代肿瘤医学》
CAS
北大核心
2022年第1期1-5,共5页
Journal of Modern Oncology
基金
辽宁省自然科学基金(编号:20170541007)。
