【摘要】 为促进紫杉醇诱导的食管癌细胞凋亡 ,探索克服耐药、提高紫杉醇疗效的新途径 ,构建了bax基因可诱导的真核表达载体 ,并用荧光素酶基因系统测定了金属硫蛋白启动子在食管癌细胞中的诱导表达及控制能力。将bax基因转入食管癌细胞Eca10 9中 ,用免疫细胞化学方法测定了Bax蛋白表达 ,并以TUNEL、DNA梯带、流式细胞术等方法分析了转染bax对紫杉醇的诱导细胞凋亡的促进作用。结果表明 ,140 μmol/LZnSO4 诱导 12h以上为最佳诱导条件 ,在此诱导条件下得到一株Bax高表达细胞株X10 97# ,在同一紫杉醇作用条件下凋亡率升高并对紫杉醇敏感度提高。提示Bax在紫杉醇诱导的细胞凋亡中起着重要作用 ,作为小分子辅助药物提高紫杉醇疗效具有很好前景更多还原

【Abstract】 An inducible mammalian expression vector of bax gene was constructed and the control ability of metallothionein II promoter in esophageal carcinoma cell line was systematically identified with luciferase report gene. After the transfection of it into human esophageal carcinoma cell line Eca109, Bax protein expression was analyzed by immunolcytochemical method. Paclitaxel induced apoptosis was determined by TUNEL assay, DNA ladder assay and flow cytometry. Results showed that 140 μmol/L ZnSO 4 for 12 h is optimal for induction of bax gene expression. Under these conditions, a clonal transfectant X1097 #, expressing bax gene effectively, was obtained. It was found that X1097 # had higher apoptotic rate and was more sensible than Eca109 upon pactaxel treatment. These results implied that Bax protein may play an important role in paclitaxel induced apoptosis. Therefore, bax protein may be promising as an helping drug to improve therapeutic effects of paclitaxel.更多还原