中药淫羊藿苷抑制肝癌HepG2.2.15细胞增殖和免疫逃逸作用研究  

Experimental study on effects of Chinese medicine ICA on the inhibition of cell proliferation and reversion of immune escape in hepatocarcinoma cell line HepG2.2.15 cells

作　　者：王谦[1] 张玲[2] 毛海婷[2] 顾洪涛[2] 夏武青[1] 温培娥[2] 李翠玲[2] 杨尚军[3] 

机构地区：[1]山东省千佛山医院,济南250014 [2]山东省医学科学院基础医学研究所山东省医药卫生肿瘤免疫与中药免疫重点实验室山东省现代医用药物与技术重点实验室,济南250062 [3]山东省医学科学院药物研究所,济南250062

出　　处：《中国免疫学杂志》2007年第10期908-911,共4页Chinese Journal of Immunology

基　　金：国家自然科学基金主任基金项目资助;No.30540041

摘　　要：目的:观察中药淫羊藿苷(ICA)对HepG2.2.15细胞增殖及对CD3AK细胞杀伤活性的影响,探讨ICA对肝癌细胞Fas/FasL途径免疫逃逸的逆转作用,为ICA的开发应用提供新的理论和实验依据。方法:MTT法检测细胞增殖和细胞杀伤活性;流式细胞术检测细胞表面分子表达水平和细胞凋亡率。结果:50μg/mlICA作用HepG2.2.15细胞48、72小时的增殖抑制作用明显,抑制率分别为22.04%、29.68%(P<0.05),呈时间依赖效应。HepG2.2.15细胞经ICA处理后,FasL的表达率由16.22%显著下降至8.29%,Fas表达率由0.79%提高到1.70%(P>0.05)。ICA可明显抑制HepG2.2.15细胞诱导Jurkat细胞凋亡,凋亡率从46.66%下降为18.20%。ICA处理HepG2.2.15细胞后,不同效靶比的CD3AK细胞的杀伤活性,可分别由对照组的15.81%、35.04%、42.85%显著提高至42.58%、67.55%、88.93%(P<0.05,P<0.01),呈效靶比依赖效应。结论:ICA能有效地抑制HepG2.2.15细胞增殖,并有一定时间依赖效应;ICA可下调FasL的表达,上调细胞表面Fas的表达,对HepG2.2.15细胞诱导的T淋巴细胞凋亡作用有一定阻断,逆转肿瘤细胞的免疫逃逸作用;ICA显著增强HepG2.2.15细胞对CD3AK细胞杀伤的敏感性。Objective:To study the effects of ICA on HepG2.2.15 cell proliferation, their sensitivity to the lysis by CD3AK effector cell, to investigate the reversal action of ICA on hepatocarcinoma cells from immune escape through Fas/FasL pathway.To provide the theoretical and experimental bases for ICA development.Methods:MTT assay was used to detect cell proliferation and CD3AK cells cytotoxicity activity;flow cytometry assay was used to examine expression of surface molecules and apoptosis rate of HepG2.2.15 cells.Results:When HepG2.2.15 cells line was treated with 50 μg/ml ICA,a significant reduction of the rate of cell proliferation was observed. Inhibition rate at 48h was 22.04%,and 29.68% at 72h.Kinetic study showed that inhibition of cell proliferation was time dependent (P<0.05). FasL expression ratio of HepG2.2.15 cells decreased from 16.22% to 8.29% after being treated by ICA (P<0.05). Fas expression ratio increased from 0.79% to 1.70%(P>0.05).ICA could inhibit apoptosis of Jurkat cells induced by HepG2.2.15 cells. In the co-culture system of HepG2.2.15 cell and Jurkat T cell, apoptosis ratio of Jurkat cell was reduced from 46.66% to 18.20% by ICA (P<0.01).ICA could significantly enhance the susceptibility of HepG2.2.15 to lysis by CD3AK. The lysis activity was upforward from 15.81%,35.04%,42.85% to 42.58%,67.55%,88.93% after ICA treatment (P<0.05,P<0.01).Conclusion:ICA could significantly inhibit HepG2.2.15 cell proliferation in certain time-dependent effect. ICA could inhibit the expression of FasL on HepG2.2.15 cell,block the way that HepG2.2.15 cells induced Jurkat cell apoptosis, and reverse tumor escape of HepG2.2.15 cells. ICA could significantly enhance the susceptibility of HepG2.2.15 cells to lysis by CD3AK.

关 键 词：淫羊藿苷 HEPG2.2.15 JURKAT细胞 FAS/FASL CD3AK 免疫逃逸 

分 类 号：R730.3[医药卫生—肿瘤]