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应用流式细胞测量术、蛋白斑点杂交及Northern blot分析在蛋白和RNA水平研究了不同浓度γ-干扰素(IFN-γ)作用48小时及去除IFN-γ作用后48小时的喉鳞癌细胞系(Hep-2)细胞组织相容性抗原(HLA)的表达情况。发现不经IFN-γ处理的对照组的Hep-2细胞表达HLA-Ⅰ类抗原,不表达Ⅱ类抗原。加IFN-γ作用48小时后HLA-Ⅰ类抗原表达增强,与IFN-γ的剂量呈量效关系,HLA-Ⅱ类抗原可被诱导表达;Hep-2细胞HLA-B27位点基因表达mRNA增强,HLA-DRα位点基因可被诱导表达mRNA,而IFN-γ作用后HLA-A2位点基因表达mRNA的量无变化。去除IFN-γ48小时后HLA抗原及mRNA的表达水平均恢复至未经IFN-γ处理的对照组水平。IFN-γ对Hep-2细胞HLA-Ⅰ类基因B位点的调节作用强于A位点,提示在HLA基因导入肿瘤细胞进行基因治疗时,用易调节表达增强的HLA-B位点基因为宜。
Flow cytometry, dot blot hybridization and Northern blot analysis were used to study the effect of different concentrations of interferon-γ (IFN-γ) for 48 hours and the laryngeal squamous cell carcinoma cell line 48 hours after IFN-γ treatment at protein and RNA levels (Hep-2) cell histocompatibility antigen (HLA) expression. Hep-2 cells in the control group that were not treated with IFN-γ were found to express HLA class I antigens but not class II antigens. After 48 hours, the expression of HLA class I antigens increased with the dose of IFN-γ, and the expression of HLA class II antigens was induced. The mRNA expression of HLA-B27 in Hep-2 cells increased , The HLA-DRα gene can be induced to express mRNA, while the amount of mRNA of HLA-A2 locus does not change after IFN-γ treatment. After 48 hours of IFN-γ removal, the expression levels of HLA antigens and mRNA were restored to the control group without IFN-γ treatment. The regulatory effect of IFN-γ on B-site of HLA-Ⅰ gene in Hep-2 cells was stronger than that of A site, which suggested that HLA-B gene with enhanced expression of HLA-B could be should.