绒毛膜癌耐药细胞系的建立及人白细胞介素2基因转染后对其多药耐药性的逆转作用  

Establishment of the drug resistant cell line of choriocarcinoma and the reversal of drug resistance by transfection of human interleukin 2 gene

作　　者：崔竹梅[1] 向阳[1] 杨秀玉[1] 刘芝华[2] 

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 [2]中国医学科学院中国协和医科大学北京协和医院肿瘤研究所分子肿瘤国家重点实验室,100730

出　　处：《中华妇产科杂志》2001年第9期549-553,共5页Chinese Journal of Obstetrics and Gynecology

基　　金：国家自然科学基金资助项目 ( 30 0 0 0 179) ;卫生部优秀青年科技人才专项基金资助项目 ( 990 1)

摘　　要：目的　建立绒毛膜癌 (绒癌 )的耐药细胞系 ,应用基因工程技术将人白细胞介素 2 (hIL 2 )基因导入绒癌耐药细胞系 ,观察hIL 2基因转染后对绒癌耐药细胞耐药性的逆转作用。方法　采用间歇诱导的方法 ,用鬼臼乙叉甙 (VP 16)诱导绒癌细胞系JEG 3 ,建立耐药细胞系 ,四甲基偶氮唑蓝比色法检测该耐药细胞系对多种化学治疗药物的耐药指数 ,及多种耐药基因包括肺耐药蛋白、多药耐药相关蛋白、谷胱甘肽转移酶、二氢叶酸还原酶和多药耐药基因 (MDR1)的mRNA表达情况。用逆转录聚合酶链反应技术 ,克隆hIL 2基因 ,将hIL 2基因插入真核表达载体pcDNA3 1( + )中 ,提取质粒 ,通过阳离子脂质体将hIL 2基因转染绒癌耐药细胞系 ,用新霉素筛选含hIL 2DNA片段的单个细胞克隆 ,检测转染hIL 2基因后该细胞耐药指数的变化 ,测定转染后细胞hIL 2和多种耐药基因mRNA的表达。结果　用VP 16采用间歇诱导法 ,成功建立绒癌的VP 16耐药细胞系JEG 3 /VP 16,并检测出MDR1mRNA表达。转染hIL 2基因后 ,绒癌细胞中检测出hIL 2mRNA表达 ,转染hIL 2基因的细胞的耐药指数降低 ,JEG 3 /VP 16细胞转染pcDNA3 1( + ) hIL 2后 ,对VP 16的耐药指数由 3 8 7降至 6 0～6 1,对甲氨蝶呤的耐药指数由 14 5降至 2 6～ 2 5 ,对更生霉素的耐药指数由Objective To establish the drug resistant cell line of choriocarcinoma and to study the transfection of the human interleukin 2 (hIL 2) gene into the established drug resistant cell line and investigate the reversal of the multidrug resistance Methods The resistant cell line was established by pulse exposed choriocarcinoma cell line JEG 3 to etopside (VP 16) for ten months The recombinant plasmid containing pcDNA3 1(+) hIL 2 gene was constructed The drug resistant cell line was transfected with the constructed plasmid by lipofectin, and the tumor cell colonies containing the IL 2 sequence were selected by genetin The expression of hIL 2 and drug resistant related genes was detected by reverse transcript polymerase chain reaction The chemosensitivity of the gene transfected tumor cells and the non transfected cell lines to methetraxate, VP 16, kengshengmycine, paclitaxol and 5 fluorouracil was determined by the methyl thiazolyl tetrazolium cytotoxicity assay Results The transfected cells expressed human hIL 2 gene, and showed the reversal of multidrug resistance by methyl thiazolyl tetrazolium assay The transfected cells expressed no multidrug resistance gene 1 (MDR1) on mRNA level Drug resistance index to VP 16 decreased from 38 7 to 6 0 and 6 1, the index to methetraxate decreased from 14 5 to 2 6 and 2 5, to methetraxate from 13 0 to 2 0 Conclusion The transfection of hIL 2 gene into the drug resistance cell line of choriocarcinoma can modulate the MDR1 expression on the mRNA level, and reverse the drug resistance

关 键 词：绒毛膜癌 转染 白细胞介素2 多药抗药性 

分 类 号：R737.33[医药卫生—肿瘤]