转染肿瘤坏死因子-α及MDR1反义RNA逆转乳腺癌多药耐药的研究

Reversal of multidrug resistance by transfection of tumor necrosis factor α and MDR1 antisense RNA into multidrug resistant breast cancer cell line

目的:转染肿瘤坏死因子-α(TNF-α)cDNA和多药耐药基因(MDR1)的反义RNA到乳腺癌耐药细胞株MCF-7/ADR中进行表达,并观察它们在乳腺癌耐药逆转中的作用。方法:应用RT-PCR和DNA重组技术构建反义绿色荧光蛋白pEGFP-MDR1融合蛋白表达载体和红色荧光蛋白pDsRed2-TNF-α融合蛋白表达载体,分别和同时导入乳腺癌耐药细胞株MCF-7/ADR中进行表达,检测转染前后细胞的生长曲线、细胞凋亡程度、MDR1-mRNA和P糖蛋白(P-gp)表达情况及对ADR敏感性的变化。结果:转染后的细胞生长明显减慢,凋亡率显著增加,MDR1-mRNA和P糖蛋白(P-gp)表达明显降低,对ADR的耐药性明显下降,敏感性增加。结论:联合运用不同的逆转耐药机制,将TNF-αcDNA及MDR1反义RNA分别或同时导入乳腺癌耐药细胞中,能有效达到逆转耐药的目的。

AIM： To study the reversal effects of muhidrug resistance by transfecting tumor necrosis factor a （TNF -a） eDNA and multidrug resistant 1 （MDR1） gene antisense RNA into multidrug resistant breast cancer cell line MCF - 7/ADR. METHODS ： The recombinant vector of enhanced green fluorescent protein （EGFP） with MDR1 antisense RNA and recombinant vector of red fluorescent protein （DsRed2） with TNF -a eDNA were constructed by RT-PCR and DNA recombinant techniques. The recombinant vectors were transfected into multidrug resistant breast cancer cell line MCF - 7/ADR. The cell growth curves, cell apoptosis rates, MDR1 gene expression at mRNA and P-gp levels, and the sensitivity to ADR were determined before and after the transfection. RESULTS： After the transfection, cells showed lower growth rate, higher apoptosis rate, lower MDR1 expression at mRNA and P-gp levels, and the sensitivity to ADR increased significantly. CONCLUSION： Transfection of TNF -a eDNA and MDR1 antisense RNA into multidrug resistant breast cancer cells may have good effects on reversal of muhidrug resistance.