中国病理生理杂志2012，Vol.28Issue(4)：613-618,6.DOI:10.3969/j.issn.1000-4718.2012.04.007
肝癌Hep3B细胞HLA-G过表达对NK细胞体外杀伤作用的影响
Effect of overexpression of human leukocyte antigen-G in hepatocellular carcinoma Hep3B cells on killing activity of NK cells in vitro
摘要
Abstract
AIM: To investigate the influence of up - regulation of human leukocyte antigen - G ( HLA - G) expression in hepatocellular carcinoma Hep3B cells on the killing effect of NK cells in vitro. METHODS; The following groups were set up: normal control group (without intervention , Hep3 B group) , empty vector control group (transfection of empty plasmid , Hep3B - GFP group) , and experimental group (transfection of plasmid pEGFP - C1 - HLA - G, Hep3B - HLA - G group). The pEGFP - Cl - HLA - G plasmid was constructed and transfected into Hep 3B cells. The mRNA and protein levels of HLA - G were detected by real - time PCR and Western blotting. The cytotoxicity of NK cells against the Hep3B cells that overexpressed HLA - G, and against the target cells that the action sites on the surface of the cells were blocked by HLA - ABC antibody were analyzed. RESULTS: The results of gel electrophoresis and sequencing showed that the inserted sequence was identical to the one we designed and had no aberrations such as mutation , deletion or insertion. After transfection of recombinant plasmid , overexpression of HLA - G in Hep3B cells was confirmed by real -time PCR and Western blotting (P <0. 01). Hep3B cells transfected with pEGFP - C1 - HLA - G showed lower cyyolytic activity. After the corresponding sites of the target cells were blocked by HLA - ABC antibody , the cytolytic activity of NK cells against Hep3B, Hep3B - GFP and Hep3B HLA - G cells was increased (P < 0. 01). CONCLUSION: The pEGFP - Cl - HLA - G was constructed in vitro and HLA - G was overexpressed at mRNA and protein levels in Hep 3B cells. Overexpression of HLA - G decreases NK cytolysis against Hep3B cells. Blockage of the sites on the surface of the target cells by HLA - ABC antibody universally enhances the killing effect of NK cells on the target cells .关键词
人类白细胞抗原-G/基因重组/肝肿瘤Key words
Human leukocyte antigen-G/Gene recombination/Liver neoplasms分类
医药卫生引用本文复制引用
曾宪成,张彤,李华,方天翎,聂常富,陈规划..肝癌Hep3B细胞HLA-G过表达对NK细胞体外杀伤作用的影响[J].中国病理生理杂志,2012,28(4):613-618,6.基金项目
国家重点基础研究发展规划(国家973规划)资助项目(No.2009CB522404) （国家973规划）
教育部博士点基金(No.20100171120087) （No.20100171120087）
广东省自然科学基金资助项目(No.10451130001004472) （No.10451130001004472）
增城市科技创新扶持基金资助项目(No.ZC201004) （No.ZC201004）
