摘要
目的分析结肠癌耐药株SW1116/HCPT(羟基喜树碱)肿瘤耐药相关基因的表达情况。方法对本所新近构建的结肠癌耐药株SW1116/HCPT的肿瘤药物耐受功能分类基因芯片的检测结果进行验证;以实时荧光定量-PCR法检测基因芯片中部分表达有改变的基因;以western blot法分析SW1116/HCPT耐药细胞株与原代细胞株间在p21和MRP2蛋白表达水平上的变化。结果p21WAF1/CIP1、MRP2(ABCC2)、BRCA2、CYP3A5等部分表达有改变基因的实时荧光定量-PCR鉴定结果与基因芯片结果基本一致;Western blot检测的p21WAF1/CIP1和MRP2(ABCC2)蛋白表达水平与其转录水平表达相吻合。结论基因芯片技术是一种大规模检测多种基因表达的有效方法,新建的结肠癌细胞耐药模型SW1116/HCPT中存在多类肿瘤耐药相关基因在转录水平和翻译水平的表达差异。
Objective ：To investigate the expression profiles of drug resistance-related genes in hydroxycamptothecin-resistant human colonic cancer SW1116/HCPT cell line. Methods-The drug resistance-related genes of our recently established hydroxycamptothecin-resistant human colonic cancer SW1116/HCPT cell line were indentified by focused DNA microarray. The changes of gene expres- sion were further verified by real-time fluorogenetic quantitative polymerase chain reaction （RFQ-PCR）. The expressions of p21 and multidrug resistance associated protein 2 （ MRP2 ） were analyzed by western blotting in both SW1116/HCPT cell line and its parent cell line. Results： Fluorescence real-time PCR confirmed the changes of p21^WAF1/CIP1, MRP2（ ABCC2）, BRCA2, and CYP3A5 detected by focused cDNA microarray. Western blot showed that the expressions of p21^WAF1/CIP1 and MRP2 proteins at translational level were coincided well with their mRNA expressions at transcriptional level. Conclusion：The cDNA microarray is an effective method for the large-scale comparison of multiple gene expressions. The expressions of many drug resistance-related genes are changed at both transcriptional and translational levels in our recently established hydroxycamptothecin-resistant human colonic cancer SW1116/HCPT cell line.
出处
《肿瘤》
CAS
CSCD
北大核心
2007年第2期96-100,共5页
Tumor
基金
上海市重点学科建设项目(编号:Y0205)
