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目的 观察TNF α基因联合异搏定 (VRP)、三苯氧胺 (TAM)逆转多药耐药性 (MDR)的效果。方法 以重组逆转录病毒为载体将TNF α基因导入具MDR表型的人乳腺癌细胞系MCF7/ADR ,经G418抗性筛选获阳性克隆MCF7/ADR TNF。以PCR与ELISA法检测目的基因的整合与表达。MTT法检测外源性TNF -a基因联合VRP、TAM的逆转MDR作用 ,应用公式I=d/D1+d/D2 分析联合逆转效应。结果 MCF7/ADR -TNF细胞中有TNF α基因的整合和表达 ,TNF α分泌量为 1737pg/ml(10 6cells/ 48h)。与阴性对照细胞相比 ,MCF7/ADR -TNF细胞对ADR的耐药性明显降低 (P <0 .0 5 ) ,耐药逆转倍数为 1.6倍。TNF α基因与VRP联合呈拮抗效应 ,而TNF α基因与TAM (6 μmol/L)联合对MCF7/ADR的耐药性逆转有明显的协同效应 (效应指数I=0 .6 4)。结论 外源性TNF α基因联合TAM具协同的逆转MDR效应
Objective To observe the effect of TNFα gene combined with verapamil (VRP) and tamoxifen (TAM) on reversal of multidrug resistance (MDR). Methods Recombinant retrovirus was used as vector to introduce TNF α gene into human breast cancer cell line MCF7/ADR with MDR phenotype. Positive clone MCF7/ADR TNF was obtained by G418 resistance screening. The integration and expression of the target gene were detected by PCR and ELISA. The effect of exogenous TNF-a combined with VRP and TAM on the reversal of MDR was determined by MTT assay. The effect of combined reversal was analyzed using the formula I=d/D1+d/D2. Results The integration and expression of TNFα gene in MCF7/ADR-TNF cells was detected. The secretion of TNF α was 1737pg/ml (106 cells/48h). Compared with the negative control cells, the resistance of MCF7/ADR-TNF cells to ADR was significantly reduced (P < 0.05), and the fold reversal of drug resistance was 1.6 times. The combination of TNFα gene and VRP showed an antagonistic effect, while TNFα gene combined with TAM (6 μmol/L) had a significant synergistic effect on the reversal of drug resistance of MCF7/ADR (effect index I = 0.64). Conclusion Exogenous TNF α gene combined with TAM has a synergistic effect in reversing MDR