B7-H3在人肝癌细胞株HepG2对外周血CD8＋T细胞活化、周期及分泌IL-17调节中的作用  

The effect of B7-H3 on human hepatocellular carcinoma cell line HepG2 mediating regulation on ac- tivation,cell cycle and IL-17 secretion of human peripheral blood CD8＋T cells

作　　者：王斐斐[1] 王国艳[1] 张光波[2] 林艳华 栾希英[1] 

机构地区：[1]滨州医学院免疫教研室,烟台264003 [2]苏州大学生物技术研究所 [3]烟台市中心血站

出　　处：《中华微生物学和免疫学杂志》2012年第11期989-994,共6页Chinese Journal of Microbiology and Immunology

基　　金：山东省自然科学基金（ZR2010HL048）;国家自然科学基金（30972778）

摘　　要：目的研究B7-H3在人肝癌细胞株HepG2对人外周血CD8^＋T细胞活化、周期及IL-17分泌等调节中的作用。方法RT—PCR及FCM检测B7-H3在HepG2细胞上的表达；应用脂质体法将PGPu6／GFP／neo-B7-H3shRNA质粒转入肝癌细胞株HepG2，阻断B7-H3的表达；免疫磁珠分选健康人外周血CD8^＋T细胞；FCM分析B7-H3分子在HepG2细胞对PHA刺激下CD8^＋T细胞活化、周期及PMA刺激下CD8^＋T细胞分泌IL—17调节中的作用。结果肝癌细胞株HepG2高表达B7-H3分子，PGPU6／GFP／neo-B7-H3shRNA质粒能有效阻断B7-H3在HepG2细胞上的表达；FCM分析结果显示，肝癌细胞株HepG2对CD8^＋T细胞活化及周期均有抑制作用；阻断B7-H3的表达后，明显减弱HepG2细胞对CD8^＋T细胞早期活化表型CD69表达的抑制作用，且能够通过下调CD8^＋T细胞G0／G1期细胞数量，上调S期细胞数量逆转HepG2细胞对CD8^＋T细胞周期的阻滞作用；在HepG2存在条件下，CD8^＋T细胞对IL—17的分泌明显增加，阻断B7-H3的表达后，IL-17的分泌被进一步上调。结论HepG2细胞高表达R7-H3分子；B7-H3能够协同HepG2细胞对CD8^＋T细胞活化表型CD69的表达及细胞周期的抑制作用；HepG2细胞上调CD8^＋T细胞对IL-17的分泌作用，但B7-H3可抑制该上调作用。Objective To investigate the effect of B7-H3 on human hepatocellular carcinoma cell line HepG2 mediating regulation on human peripheral blood CD8 ＋T cell activation, cell cycle and secretion of IL-17. Methods The expression of the B7-H3 on HepG2 cells was detected by RT-PCR and FCM respec- tively. B7-H3 was silenced by PGPU6/GFP/neo-B7-H3shRNA plasmid via cathodolyte liposome transfection method. CD8＋T cells were sorted from healthy human peripheral blood with immunomagetie beads. The effect of HepG2 cells on activation, cell cycle and cytokine secretion of CD8＋T cells which was stimulated by PHA or PMA respectively were analyzed by FCM. Results B7-H3 was highly expressed on HepG2 cells,and PG- PU6/GFP/neo-B7-H3shRNA plasmid could effectively block down its expression. Otherwise, HepG2 cells could inhibit the expression of CD69, the early activation phenotype of T eel1, blockade B7-H3 on HepG2 cells could significantly attenuate the inhibitory effects. Likewise, blockade B7-H3 on HepG2 cells apparently reversed the inhibitory effects of HepG2 ceils on CD8＋T cell cycle through down-regulating the cell number in G0/G1 phase and up-regulating the cell number in S phase;Moreover, HepG2 cells caused a sharp increase of IL-17 which was secreted by CD8＋T cells and the level of IL-17 was further up-regulated after blocking down B7-H3. Conclusion HepG2 ceils highly expressed BT-H3 that could promote the inhibitory the effect of HepG2 on expression of CD69 and cell cycle of CD8＋T cells. HepG2 cells were able to up-regulate the lev- el of IL-17 secreted by CD8＋T cells ,in which B7-H3 played an inhibitory role.

关 键 词：人肝细胞癌 HepG2细胞 CD8^+T细胞 B7-H3 

分 类 号：R735.7[医药卫生—肿瘤]