CAJ | 학술논문

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HepG2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。
목적：탐토살상세포면역구단백양수체（KIR）기인재간암세포면역살상중적작용。방법외주혈단개핵세포（PBMC）여간암세포HepG2안불동효파비공부육，관찰불동효파비공부육하PBMC중KIR기인가족적표체、γ간우소（IFN-γ）분비、간암세포형태학개변급PBMC대간암세포적살상정황。살상솔비교채용단인소방차분석화LSD-t검험。결과여간암세포공부육12 h후PBMC활화성KIR기인표체개시증가，24 h후하강。억제성KIR기인공부육12 h후개시하강。보조활화기인DAP12일직보지고표체수평。PBMC중IFN-γ적함량수착효파비적승고이강저，12 h체도고봉。공부육후불동효파비조적간암세포균표현위염색질농축，세포핵정반구상혹반월형상차변연화적세포비례증가，세포정지왕성분렬。효파비1∶1조적상대살상솔위（8±3）%，10∶1조위（14±4）%，50∶1조위（32±6）%，50∶1조명현고우1∶1조화10∶1조（LSD-t=5.97，4.61；P<0.05）。결론활화성KIR기인재간암세포면역살상중발휘중요작용。
ObjectiveTo investigate the effect of killer cell immunoglobulin-like receptor (KIR) gene in immune killing of hepatoma cells.MethodsPeripheral 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.ResultsThe expression of activating KIR gene increased after 12 h of co-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).ConclusionThe activating KIR gene plays an important role in immune killing of hepatoma cells.