摘要
目的 建立多药耐药基因 (MDR1)稳定表达的人肝癌Bel 740 2耐药细胞株 ,为应用RNA干扰技术逆转肿瘤多药耐药基因的表达提供实验模型。方法 应用阿霉素 (ADM )长期培养筛选建立Bel 740 2耐药细胞株 ,应用MTT法、流式细胞仪(FCM )及逆转录聚合酶链反应 (RT PCR )对该细胞株相关特性 (耐药范围、致死剂量、p 糖蛋白功能及其表达阳性率、MDR 1基因检测 )进行比较研究。结果 经MTT法检测 ,耐药细胞株耐药性明显增高 ,FCM检查发现从敏感细胞到耐药细胞 ,其阿霉素潴留功能由 77.7%降至 2 5 .1% ,P 糖蛋白 (P gp)阳性率由 1.4%升高至 5 4.0 % ,RT PCR检查显示该细胞中MDR1mRNA呈高表达。结论 成功建立了稳定表达MDR1基因的人肝癌Bel 740 2耐药细胞株 ,该细胞中P gp呈高表达 。
Objective To establish Multi-drug resistant strain stably expressing human hepotacarcinoma Bel-7402 and to study its multi-drug resistance mechanism to provide laboratory models for application of RNA interfere technique in the fight against drug resistancetumors.Methods Bel-7402 Multi-drug resistant strain was established by long -term culturing and screening of adriamycin.And the following techniques,namely,MTT,FCM and RT-PCR were used systemically the relevant features of the strain,including extent of drug tolerance,lethal dose,function and positive rate of P-gp and test of gene MDR1.Results MTT showed that the Multi-drug resistance of strain increased significantly FCM found,From sensetive cell to Multi-drug resistance cell.With the function of P-gp falling from 77.7% to 25.1% and positive rate of P-gp rising from 1.4% to 54.0%.PT-PCR revealed that Multi-drug resistance cell strain had MDRmRNA over expression.Conclusion Multi-drug resistant strain that stably express human hepatocacinoma Bel-7402 cell was successfully established.The strain has high expression of P-gp and high stability.
出处
《实用癌症杂志》
2003年第6期580-582,共3页
The Practical Journal of Cancer
关键词
MDR1基因
稳定表达
肝癌
多药耐药细胞株
Multi-drug resistant strain
Human hepatocarcinoma Bel-7402 cell
MDR1 gene
