HLA-B7可诱导启动子调控TNF-α和IFN-β基因协同增强抗肿瘤效应  

The co-expression of TNF-α, IFN-β gene controlled by HLA-B7 promoter enhances antitumor effect

作　　者：杨淑英[1] 段芳龄[1] 潘卫[2] 曹广文[2] 高天慧[1] 陈秋丽[2] 郭兴中[2] 李玉光[2] 戚中田[2] 

机构地区：[1]郑州大学消化疾病研究所 [2]第二军医大学基础部微生物教研室

出　　处：《中华微生物学和免疫学杂志》2003年第4期275-278,共4页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金资助项目 ( 39970 819;2 0 170 5 14 )

摘　　要：目的　将TNF α和IFN β基因导入人肝癌细胞株Liu6和SMMC 772 1,以增强其免疫原性。方法　以腺病毒为载体 ,以IRES连接TNF和IFN基因 ,选择HLA B7可诱导启动子调控其表达。分别用Southern Northernblot和MTT法检测TNF α和IFN β基因的整合、转录与表达。流式细胞仪检测MHCⅠ类分子表达。CTL细胞毒性实验检测T细胞对转染细胞的杀伤作用。细胞增殖实验检测目的基因的表达对肿瘤细胞的抑制作用。结果　TNF α和IFN β基因可整合入转染细胞 ,并被有效转录。转染细胞MHCⅠ类分子的表达比对照组高 3～ 4倍。TNF活性 :Liu6细胞为 3 .778ng ml,SMMC 772 1为2 .0 17pg ml;2 4hIFN最高活性分别为 397.8U 10 6 个细胞和 345 .8U 10 6 个细胞。对照组与实验组在效靶比分别为 5∶1,10∶1,15∶1时 ,CTL细胞毒性实验结果差异均有显著性 (P <0 .0 5 ) ,表明T细胞杀伤作用明显增强。细胞增殖实验结果显示实验组与对照组差异有显著性 (P <0 .0 5 )。目的基因的表达对肿瘤细胞具有一定的抑制作用 ,抑制率分别为 5 .5 %(Liu6 ) ,3.7%(SMMC 772 1)。结论　人TNF α和IFN β基因导入人肝癌细胞株Liu6和SMMC 772 1可明显增加其对TNF、IFN和MHCⅠ类分子的表达 ,增强机体对转染瘤细胞的杀伤作用。Objective TNF-α and IFN-β genes were introduced into human hepatocellular carcinoma cell line Liu6 and SMMC-7721 to increase tumor immunogenicity. Methods Recombinant adenovirus was used as vector, TNF-α and IFN-β were connected through IRES and HLA-B7 promoter was selected to control their expression. Southern/Northern blot was used to detect the transfected genes expression. MHC class Ⅰ molecule expression was measured by FACS assays. MTT was used to detect TNF-α and IFN-β activity, cytotoxic activity of CTL and the growth of tumor cell. Results TNF-α and IFN -β gene expression were verified, MHC class Ⅰ molecule expression in test groups was much higher than that in control group. TNF-α activity was 3.778ng/ml, 2.017ng/ml in Liu6 and SMMC-7721 respectively, IFN-β activity was 397.8U/24?h per 106 cells and 345.8 U/24h per 106 cells. When the cytotoxic activity of CTL was compared between test group and control group, there was a significant difference (P<0.05) at effector: target cell ratios of 5∶1, 10∶1, 15∶1 that suggested an enhanced lysis effect of CTL. The expression of target gene could resist the tumor cell growth. The resistant rate is 5.5% (Liu6), 3.7% (SMMC-7721) respectively. Conclusion The transfected human hepatocelullar carcinoma cells had an elevated immunogenicity that could be explained by production of TNF, IFN and MHC class Ⅰ molecule. [

关 键 词：HLA—B7 诱导 启动子 调控 TNF-Α IFN-Β 基因协同增强抗肿瘤效应 基因治疗 

分 类 号：R730.3[医药卫生—肿瘤]