杀伤细胞免疫球蛋白样受体基因在肝癌细胞免疫杀伤中的作用  

Effect of killer cell immunoglobulin-like receptor gene in immune killing of hepatoma cells

作　　者：何免[1] 邱小惠[1] 张文峰[1] 沈晗[1] 王辉[1] 薄华本[1] 黄树林[1] 曾宪成 邵红伟[1] 

机构地区：[1]广东药学院生命科学与生物制药学院广东省生物技术候选药物研究重点实验室,广州510006 [2]广州市增城区人民医院普通外科,513000

出　　处：《中华肝脏外科手术学电子杂志》2015年第5期318-322,共5页Chinese Journal of Hepatic Surgery(Electronic Edition)

基　　金：国家自然科学基金基金(31100664;31300737;81303292);广东省自然科学基金(10151022401000024;2014A030313586);广东药学院师资队伍建设专项经费(20080927);广州市卫生局项目(20141A011117;20151A011112);广州市科技计划项目(2013J4100081)

摘　　要：目的探讨杀伤细胞免疫球蛋白样受体(KIR)基因在肝癌细胞免疫杀伤中的作用。方法外周血单个核细胞(PBMC)与肝癌细胞Hep G2按不同效靶比共孵育,观察不同效靶比共孵育下PBMC中KIR基因家族的表达、γ干扰素(IFN-γ)分泌、肝癌细胞形态学改变及PBMC对肝癌细胞的杀伤情况。杀伤率比较采用单因素方差分析和LSD-t检验。结果与肝癌细胞共孵育12 h后PBMC活化性KIR基因表达开始增加,24 h后下降。抑制性KIR基因共孵育12 h后开始下降。辅助活化基因DAP12一直保持高表达水平。PBMC中IFN-γ的含量随着效靶比的升高而降低,12 h达到高峰。共孵育后不同效靶比组的肝癌细胞均表现为染色质浓缩,细胞核呈半球状或半月形状且边缘化的细胞比例增加,细胞停止旺盛分裂。效靶比1∶1组的相对杀伤率为(8±3)%,10∶1组为(14±4)%,50∶1组为(32±6)%,50∶1组明显高于1∶1组和10∶1组(LSD-t=5.97,4.61;P<0.05)。结论活化性KIR基因在肝癌细胞免疫杀伤中发挥重要作用。Objective To investigate the effect of killer cell immunoglobulin-like receptor(KIR) gene in immune killing of hepatoma cells.Methods Peripheral blood mononuclear cell(PBMC) and hepatoma cells were co-cultured with different effector-target ratios.The expression of KIR gene family in PBMC,the content to interferon-γ(IFN-γ),the morphological change of hepatoma cell and the cytotoxicity to hepatoma cell by PBMC were observed after the co-incubation with different effector-target ratios.The comparison on cytotoxicity rates was conducted using one-way analysis of variance and LSD-t test.Results The expression of activating KIR gene increased after 12 h ofco-culture,but decreased after 24 h of co-culture.The expression of inhibitory KIR gene decreased after 12 h of co-culture.DAP12 maintained high expression all the time.The content of IFN-γ in PBMC decreased with the increase of effector-target ratio and reached the peak at 12 h of co-culture.Hepatoma cells co-cultured with different effector-target ratios were observed with increased chromatin condensation,rising proportion of cells with hemispherical or half moon shape and marginalized nucleus,and stagnant of active cell division.The cytotoxicity rate of effector-target ratio 1∶1,10∶1 and 50∶1 was(8±3) %,(14±4) % and(32±6) %,respectively,with 50∶1 group significantly higher than 11∶1 and 10∶1 group(LSD-t=5.97,4.61;P<0.05).Conclusion The activating KIR gene plays an important role in immune killing of hepatoma cells.

关 键 词：受体 KIR 白细胞 单核 受体 自然杀伤细胞 癌 肝细胞 干扰素Γ 

分 类 号：R735.7[医药卫生—肿瘤]