论文部分内容阅读
目的建立转染3种TNF-α基因(分泌型TNF-α突变体、跨膜型TNF-α突变体和野生型TNF-α的H22细胞株,以比较不同类型TNF-α的杀瘤效应。方法采用逆转录病毒载体,将3种TNF-α基因导入小鼠H22肿瘤细胞中,用Southern印迹、RT-PCR、流式细胞术和生物学活性检测等方法,从基因的整合、RNA转录、蛋白的表达及其生物学活性4个方面,对转染细胞进行检测鉴定。结果3种TNF-α基因均在H22细胞中得到有效表达,并具有生物学活性。结论获得3种稳定高效表达TNF-α的H22肿瘤细胞株,为进一步研究跨膜型和分泌型TNF-α体内的抑瘤效应建立了良好的实验模型。
Objective To establish a tumorigenicity effect of different types of TNF-α transfected with three TNF-α genes (secretory TNF-α mutant, transmembrane TNF-α mutant and wild-type TNF-α). METHODS: Three TNF-α genes were introduced into mouse H22 tumor cells using retroviral vectors and were subjected to Southern blotting, RT-PCR, flow cytometry, and biological activity assays, from gene integration and RNA transcription. The expression of the protein and its biological activity were tested in four aspects.Results The three TNF-α genes were effectively expressed in H22 cells and had biological activity.Conclusion Three stable and high-efficient expression of TNF were obtained. The α-H22 tumor cell line established a good experimental model for the further study of the anti-tumor effect of transmembrane and secretory TNF-α in vivo.