艾军魁 陈一戎 秦大山 孟筱琦 王兴民 王志平 白淑芳 刘国栋 200025 上海第二医科大学瑞金医院泌尿外科 中华泌尿外科杂志 1999 0 20 5


    关键词：膀胱肿瘤,癌,基因,转移 期刊 zhmnwkzz 0 论 著 fur -->


    

【摘要 】 目的 探讨肿瘤坏死因子TNF-α cDNA突变体对膀胱癌的作用。 方法 以突变的TNF-α cDNA为目的基因,pcDNA3.1(+)为载体,用磷酸钙沉淀法将其导入人膀胱癌细胞株EJ。 结果 PCR检测证实外源基因整合于EJ细胞内，放射免疫分析法检测转基因后细胞培养上清TNF-α表达量为(0.664±0.225)ng/ml,小鼠肉瘤细胞L929杀伤试验证实其生物活性为4～8U/ml，软琼脂培养细胞集落形成抑制率为36.5%。 结论 TNF-α cDNA突变体用于膀胱癌的基因治疗是可行的。

Expression and Function of TNF-αcDNA mutant in human bladder cancer EJ cell lines AI Junkui ,CHEN Yirong ,QIN Dashan ,et al .Instituteof Urology ,Second Affiliated Hospital ,Lanzhou Medical College ,Lanzhou 730030

【Abstract 】 Objective Tostudy the function of a gene mutant of tumor necrosis factor-α (TNF-α )in bladder cancer. Methods A TNF-α cDNA mutant was selectedas the target gene.The novel mutant was transduced into human bladder cancer EJ cell lineswith the plasmid pcDNA 3.1(+) as the vector by the method of calcium phosphate-DNA coprecipitation. Results Polymerase chain reaction (PCR) analysisconfirmed the exogenic gene was stably integrated into EJ cell lines.The level of TNF-αin the supernatant collected from EJ cells transfected with TNF-α cDNA mutantwas (0.664±0.225)ng/ml by radioimmunoassay (RIA) and L929 bioassay showed itscytotoxicity was 4～8U/ml.The formation ability of clony in soft agar was decreasedsignificantly and the inhibition rate was 36.5%. Conclusions The TNF-α cDNA mutant can be used in gene therapy for bladder cancer.

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