MDR1基因稳定表达的人肝癌多药耐药细胞株的建立

目的 建立多药耐药基因(MDR1)稳定表达的人肝癌Bel-7402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-7402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、MDR1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25．1％，P-糖蛋白(P—gp)阳性率由1．4％升高至54．0％，RT-PCR检查显示该细胞中MDR1 mRNA呈高表达。结论 成功建立了稳定表达MDR1基因的人肝癌Bel-7402耐药细胞株，该细胞中P—gp呈高表达，稳定性好。

Establishment of Human Hepatocarcinoma Multi-drug Resistant Strain and Stable Expression of MDR1

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.