结肠癌耐药细胞株LoVo/L-OHP的建立及其耐药机制

摘要

Objective :To estahlish a human drug resistance colon carcinoma cell line Lovo/L - OHP. and to explore its potential drug resistant mechanism. Methods : LoVo/L - OHP of a human drug - resistance colon carcinoma cell line was induced by continuously exposing human colon carcinoma cells to gradually increasing concentrations of L - OHP. The growth curve was observed. The drug resistance of LoVo/L - OHP was measured hy MTT assay and the drug resistant index ( RI ) was calculated. Several genes selected associated drug resistance genes were confirmed by reverse transcription - polymerase chain reaction ( RT - PCR ). Results : Compared with parental cells , the resistance cell line had a slower growth rate and larger morphology. RT - PCR results of LoVo/L - OHP cell line showed up - regulated P - gp, bcl - 2 , ERCC - 1genes, and p53 gene down - regulated. Conclusion : The altered biological properties of LoVo/L - OHP may he related to its drug resistance phenotype. Several genes, such as P - gp,bcl - 2, ERCC - 1 genes up - regulated and p53 gene down - regulated were possibly mechanism of drug resistance.%目的:建立获得性奥沙利铂(L-OHP)耐药的结肠癌细胞模型LoVo/L-OHP,并初步研究其耐药机制.方法:采用L-OHP浓度递增法建立人结肠癌细胞耐药模型LoVo/L-OHP,观察其生长规律并绘制细胞生长曲线;用MTT法鉴定耐药细胞株耐药性并计算耐药指数(RI);用半定量RT-PCR方法对部分耐药相关基因在耐药细胞及其亲本细胞中的表达情况进行分析.结果:成功建立了耐药的结肠癌细胞模型LoVo/L-OHP,LoVo/L-OHP细胞与LoVo细胞相比,生长缓慢,触角增多.通过RT-PCR半定量分析,P-gp、bcl-2、ERCC-1在LoVo/L-OHP中的表达上调,而p53基因表达下调.结论:LoVo/L-OHP细胞株耐药性稳定,耐药机制可能与P-gp、bcl-2、ERCC-1基因上调、p53基因下调多因素有关.