转录因子Snail促进结直肠癌细胞对奥沙利铂耐药的机制研究  

Molecular mechanism of transcription factor Snail mediated chemoresistance of colorectal cell HCT116 to Oxaliplatin

作　　者：钟文[1] 叶磊[2] 童磊 王昊[4] 

机构地区：[1]安徽省立医院病理科,安徽合肥230001 [2]安徽省立医院B超室,安徽合肥230001 [3]淮南市凤台县第一人民医院普外科,安徽淮南230032 [4]安徽省立医院检验科,安徽合肥230001

出　　处：《安徽医药》2017年第2期315-318,共4页Anhui Medical and Pharmaceutical Journal

基　　金：国家自然科学基金(81502599);安徽省自然科学基金(1608085QH217);中国博士后基金(2016M592040)

摘　　要：目的研究转录因子Snail促进结直肠癌细胞HCT116对奥沙利铂产生耐药性的分子机制。方法采用MTT法分别检测过表达Snail蛋白和空载质粒的HCT116细胞对奥沙利铂的敏感性差别,以及RNA敲除-糖蛋白(P-gp,编码基因ABCB1)蛋白的HCT116细胞对奥沙利铂的敏感性的影响。实时定量PCR方法检测过表达Snail对耐药相关基因表达的影响。蛋白质免疫印迹技术Western blot检测过表达Snail对P-gp蛋白表达的影响。结果过表达转录因子Snail可以明显促进结直肠癌细胞HCT116对奥沙利铂产生耐药性。实时定量PCR结果表明:过表达Snail明显提高了P-gp蛋白编码基因ABCB1的表达,而对其它耐药基因如ABCC1、ERCC1等没有明显影响。Western blot结果显示过表达Snail可以促进P-gp蛋白的表达。敲除P-gp蛋白可以明显提高HCT116细胞对于奥沙利铂的敏感性。结论过表达Snail可以明显促进结直肠癌细胞HCT116对奥沙利铂的耐药性,其机制与Snail促进耐药蛋白P-gp的表达有关。Objective To investigate the molecular mechanism of transcription factor Snail mediated chemoresistance of colorectal cell HCT116 to Oxaliplatin. Methods The effect of overexpression Snail or knockdown P-gp on sensibility of HCT116 cells to Oxaliplatin was analyzed by MTT assay. Real time PCR was used to detect mRNA expressions of drug resistance genes influenced by overexpression Snail. Western blotting was used to detect the effect of Snail on P-gp expression. Results Compared to vector plasmid,overexpression Snail obviously enhanced the resistance of HCT116 cells to Oxaliplatin. The results of real time PCR demonstrated that overexpression Snail significantly enhanced ABCB1 mRNA expression, which encoded P-gp protein, but not the other drug resistance genes, such as ABCC1 ,ERCC1. Western blotting result show over overexpression Snail promote the expression of P-gp protein. Importantly, knockdown P-gp expression enhanced the sensibility of HCT116 cells to Oxaliplatin. Conclusions Overexpression Snail increased the chemoresis-tance of HCT116 cells to Oxaliplatin by up-regulated multidrug resistance protein P-pg expression.

关 键 词：结直肠癌 SNAIL 耐药 奥沙利铂 P-GP 

分 类 号：R735.34[医药卫生—肿瘤]