摘要
运用小鼠白血病病毒MoMLV为载体,将TNFa基因转导致人胶质瘤细胞SHG-44,14dG418筛选出细胞克隆.检测TNFa的表达.观察细胞生长.结果:转基因细胞培养上清液中有高浓度TNFa表达分别为(5 198.7±3 757.4)pg/ml和(3 217.4±1 180.6)pg/ml(P<0.05),生物活性达320.0U/ml.“RT-PCR”检测转基因细胞有特异mRNA表达,转基因细胞生长减慢.转染TNFa基因可诱导胶质瘤细胞的生长抑制.
Human tumor necrosis factor (hTNF) α gene was transduced into human glioma cells (SHG-44) by the murine leukemia virus derived retroviral vector, to detect the expression of TNFα gene and observe the growth rates of transfected cells. A high level of human TNFαin the culture medium was determined by an enzyme-linked immunoassay method , the biological activity of hTNFα in the supernatants was detected with a bioassay and the specific expression of hTNFα mRNA in the transfected cells was detected by RT-PCR. Introduction of hTNFαgene into SHG-44 glioma cells inhibited the growth, and flow cytometry analyses showed the decreased G2M phase and increased GOG1 phase in cell cycle. The results suggest the potential use of TNFα transgene system for developing gene therapy of malignant gtlioma.
出处
《中国临床神经科学》
1998年第3期149-153,共5页
Chinese Journal of Clinical Neurosciences
