人肿瘤坏死因子-α基因转导绒癌耐药细胞系体外耐药逆转的研究  

In vitro reversal of multidrug resistance by transduction of human tumor necrosis factor-α into drug resistant cell line of choriocarcinoma

作　　者：冯凤芝[1] 向阳[1] 张卫光[2] 崔竹梅[3] 张颖[1] 杨秀玉[1] 

机构地区：[1]中国医学科学院中国协和医科大学北京协和医院妇产科,北京100730 [2]北京大学医学部解剖学与组织胚胎学系,北京100083 [3]青岛大学医学院附属医院妇科

出　　处：《肿瘤》2003年第4期283-286,共4页Tumor

基　　金：国家自然科学基金资助项目 (编号 :3 0 0 0 0 179)

摘　　要：目的　将人肿瘤坏死因子 alpha(hTNF α)基因导入已建立的绒癌耐药细胞系 ,观察hTNF α基因转导后对绒癌细胞耐药性逆转的体外作用。方法　通过阳离子脂质体将hTNF α基因转导绒癌耐药细胞系 ,用新霉素筛选含hTNF α片段的单细胞克隆 ,用RT PCR方法和细胞免疫组织化学方法 ,检测转导hTNF α基因的耐药细胞中MDR1mRNA和MDR1蛋白 (P gp)表达水平的改变。采用四甲基偶氮唑蓝比色法 ,检测转导hTNF α基因的耐药细胞对足叶乙甙 (VP 16 )的耐药指数。结果　转导hTNF α基因后 ,绒癌耐药细胞中检测出hTNF αmRNA表达 ,转导hTNF α基因在mRNA水平能一定程度的逆转绒癌耐药细胞的MDR1,而在P gp蛋白水平几乎能完全逆转绒癌耐药细胞的MDR1。转导hTNF α基因的细胞的耐药指数明显降低。结论　hTNF α基因转导后 ,可通过调节MDR1的表达 。Objective To investigate in vitro reversal of multidrug resistance of a drug resistant cell line of choriocarcinoma by transducing human tumor necrosis factor-α(hTNF-α) via transduction of hTNF-α gene into the established drug resistant cell line. Methods The drug resistant cell line of choriocarcinoma was transduced with an eukaryotic expression plasmid containinghTNF-α gene by lipofectin, and the tumor cell clones containing hTNF-α gene were selected by genetin. Expression of MDR1 mRNA of hTNF-α-transduced drug-resistant choriocarcinoma cell line was investigated by using RT-PCR, and P-glycoprotein (P-gp) expression was determined immunohistochemically with the monoclonal antibody MDR1. The chemosensitivity of thehTNF-α transduced drug-resistant choriocarcinoma cells and the non transduced cell lines to etopside (VP-16) was determined by the methyl thiazolyl tetrazolium cytotoxicity assay. Results Transduction and expression of hTNF-α in drug-resistant choriocarcinoma cell line was found to reverse MDR1 on the mRNA and P-gp levels. Drug resistance index to VP-16 in hTNF-α transduced drug-resistant choriocarcinoma cell line decreased significantly. Conclusion Transduction of hTNF-α gene into drug-resistant choriocarcinoma cell line can modulate the expression of MDR1 on the mRNA and P-gp levels, and reverse the drug resistance.

关 键 词：绒癌 人肿瘤坏死因子-Α 耐药性 基因转导 MDRl蛋白 

分 类 号：R737.33[医药卫生—肿瘤]