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目的 建立 Hep- 2 v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- 2 v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- 1 70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筛选后检测其耐药性状 ,MTT法测得其 VCR的 IC50 由筛选前的 6 0 nmol/ L上升到约 1 2 0 0 nmol/ L ,流式细胞仪检测细胞表面糖蛋白 P-1 70阳性率由亲本细胞的 2 0 %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 2 6 .3%。结论 本研究建立的耐药细胞株 Hep- 2 v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- 2 v细胞株可作为 MDR研究的良好模型
Objective To establish a Hep-2 v multidrug resistance (MDR) cell line to further investigate the characteristics and reversal of MDR in throat tumors. Methods Hep-2 cells were selected as the parent cells and screened by VCR for selection of Hep-2 v-resistant cell lines. The positive rate of the surface glycoprotein P-1 70 and the intracellular accumulation of rhodamine were measured by flow cytometry before and after screening. The cytotoxicity of VCR on the two groups of cells before and after screening was determined by MTT assay. Results The drug resistance of Hep-2 cells was detected by VCR screening. The IC50 of VCR was increased from 60 nmol / L before screening to about 120 nmol / L by MTT assay. The cell viability was detected by flow cytometry The positive rate of surface glycoprotein P-1 70 increased from about 20% of the parental cells to over 90%. Flow cytometry detection of intracellular rhodamine accumulation rate decreased from an average of 76.2% before screening to 26.3%. CONCLUSION: Hep-2 v cell line established in this study is mainly characterized by mdr1 gene-mediated MDR and its sensitivity to VCR is obviously lower. Therefore, Hep-2 v cell line can be used as a good model for MDR study