【摘要】 Hp SV2 - neo质粒作为载体 ,在 Eco RI酶切位点上插入小鼠金属硫蛋白 (MT- I)基因 ,然后用 DNA-磷酸钙介导的基因转移法将重组质粒转染 He L a细胞 ,经筛选得到表达 MT- I的抗性细胞克隆。经检测 ,G418抗性细胞内有 MT的特异性表达 ,表达量是未转染细胞的 2 .6倍。用不同浓度顺铂、阿霉素处理细胞 ,观察 MT- I基因的表达与 He L a细胞耐药性的关系。结果表明 :0 .1μmol/ ml顺铂作用细胞 72 h后 ,顺铂对转染和未转染 MT细胞的生长抑制率分别为 34%和 82 % (P<0 .0 5 ) ;半数细胞生长抑制浓度 (IC5 0 )分别为0 .144和 0 .0 6 1μmol/ ml,二者相差 2 .36倍。用 0 .2μmol/ ml阿霉素作用细胞 72 h后 ,对转染和未转染细胞的生长抑制率分别为 18%和 2 5 % ,差异无显著性 (P>0 .0 5 )。结果表明 ,MT与肿瘤细胞的耐药性有一定相关性更多还原

【Abstract】 Metallothionein I(MT I) gene was inserted into EcoRIsite by using pSV2 neo plasmid vector. Recombiant plasmid was transfected into HeLa cells by DNA calcium phosphate precipitation technique. MT I expression colones were growing in medium including G418. The amount of MT I expression in transfected cells was found 2 6 times higher than that of non transfected ones. In order to observe the relationship between the expression of MT I gene in cells and drug resistance, cells were treated with different concentrations of cisplatin and adriamycin respectively. The results indicated that cisplatin(0 1μmol/ml) inhibited the growth of both transfected and non transfected cells. The inhibitory rates were 34% and 82% respectively( P <0 05). IC 50 (50% inhibitory concentration for cell growing) was 0 144μmol/ml and 0 061 μmol/ml and the ratio of them was 2 36: 1 after the treatment of cisplatin 72h later. The cells were treated with adriamycin 72h later, the inhibitory rates of transfected and non transfected cells were 18% and 25% separately. The rates showed no significant difference ( P>0 05 ). The results indicated that MT was related to drug resistance of tumor cells. WTHZ更多还原