膜结合型肿瘤坏死因子在人乳腺癌细胞中的转染  

MEMBRANE ASSOCIATED TUMOR NECROSIS FACTOR-α TRANSFECTED INTO HUMAN BREAST CANCER CELLS

作　　者：王桂林[1] 谢军[1] 荀雅书[2] 刘肇宁[1] 许佐良 

机构地区：[1]内蒙古医学院药理教研室 [2]内蒙古医学院分析化学教研室 [3]北京市肿瘤研究所,呼和浩特010050

出　　处：《内蒙古医学院学报》2000年第4期215-219,共5页Acta Academiae Medicinae Neimongol

摘　　要：目的 :研究膜结合型肿瘤坏死因子α(TNFα)在基因转染中的特点及其在肿瘤基因治疗中的作用等特性。方法 :用反转录病毒为载体 ,将含有突变的膜结合型TNFα的质粒 ,用lipofectin法转入包装细胞株crip细胞。又用收集到的病毒上清感染人乳腺癌细胞MDA2 31,采用G418筛选 ,DNA印迹法和聚合酶链反应方法鉴定该基因转染的结果。结果 :经上述方法鉴定表明该外源基因已成功转染人乳腺癌细胞中 ,为进一步研究膜结合型TNFα的功能奠定了基础。结论 :经初步研究表明 ,含有膜结合型肿瘤坏死因子的质粒可以通过lipofectin法成功导入crip细胞 。Objective:To research about the characteristics of membrane associated tumor necrosis factor in gene transfection and its function in gene therapy of cancer.Methods:A double-copy colony leukemia virus-based retroviral construct containing both the neo gene and a membrane associated tumor necrosis factor cDNA was used to transfer into a packaging cell line crips with lipofectin procedure.Then the supernatant of the virus was collected and its titer was assayed by counting the number of colony-forming cells using NIH 3T3 cells.And then human breast cancer cells were infected with the virus supernatant in co-culture way.Evidences for integration of the cDAN into the human genomic DAN were obtained by the ways of screening with G418,Southern blotting and polymerase chain reaction amplification.Results:The results show that the cDAN was integrated into human genomic DNA.The PCR product was detected and also show the integration of the foreign gene into human genomic DNA.Conclusion:The foreign gene containing membrane associated tumor necrosis factor gene and neo gene had been transfected into human breast cancer cells and integrated into genomic DNA of MDA 231 cells successfully.

关 键 词：膜结合型肿瘤坏死因子 基因 乳腺癌细胞 基因转染 基因治疗 

分 类 号：R737.905[医药卫生—肿瘤]