下咽癌Hep—2v多药耐药细胞株的筛选

目的 建立Hep—2v多药耐药(MDR)细胞株，以进一步研究探讨咽喉部肿瘤MDR特征及其逆转方法。方法 以下咽癌Hep—2细胞为亲本细胞，用长春新碱(VCR)进行耐药细胞筛选，选择出Hep—2v耐药细胞株。用流式细胞仪测定筛选前后两组表面糖蛋白P-170的阳性率及细胞内罗丹明蓄积率。以MTT法测定筛选前后VCR对两组细胞的细胞生长半数抑制浓度(IC50)比较其细胞毒作用。结果 亲本细胞Hep—2经VCR筛选后检删其耐药性状，MTT法测得其VCR的IC50由筛选前的60nmol／L上升到约1200nmol／L，流式细胞仪检测细胞表面糖蛋白P—170阳性率由亲本细胞的20％左右增加到90％以上。流式细胞仪检测细胞内罗丹明蓄积率从筛选前的平均76．2％降至26.3％。结论 本研究建立的耐药细胞株Hep—2v主要表现为mdr1基因介导的MDR性状，其对选择药物VCR的敏感性明显降低，因此Hep—2v细胞株可作为MDR研究的良好模型。

Hep-2v Multidrug-Resistance Cell Line selected by Vincristine and its identification

Objective Establishing Hep 2v Cell Line to study Multidrun Resistance(MDR) and its reversion.Methods Establishment of resistance cell line:The parental cell from pharyngeal cancer exposed to vincristine(VCR)with increasing concentration for four months.The selected cell,termen Hep 2v,was as a subject for this study.Quantitative analysis of P 170 was made by Flow Cytometry(FCM).The drug accumulation in the cells was measured by FCM after Rhodanmin staining,and the cellular toxicity of VCR was determined by MTT method.Results Selected cell with VCR appeared MDR phenotype obviously.The positive rate of P 170 was from about 20% in parental cell Hep 2 up to more than 90% in selected cell Hep 2v.Correspondingly,the drug accumulation in the cells indicated by Rhodamin staining decreased from 76 2% down to 26 3% after stressed by VCR.The IC 50 of VCR was from 60 nmol/L in Hep 2 up to 1200 nmol/L in Hep 2v.Conclusion The cell line Hep 2v established in this study revealled the mdrl mediated MDR phenotype.The sensitivity of the cell to VCR was decreased greatly.It is indicated that Hep 2v can be served as a model for the study of MDR.