LIGHT-INF-γ基因配比转染HepG2细胞对其凋亡及Fas-FasL系统表达的影响.docVIP

LIGHT/INF-γ基因配比转染HepG2细胞对其凋亡及Fas/FasL系统表达的影响 作者：王正华　吴力群　韩冰　　 卢云　吕振华 刘相萍　杨堃　隋爱华　毕崇尧　李金朋 【摘要】 目的：观察脂质体介导的LIGHT和IFN-γ配比转染HepG2细胞后对其凋亡及Fas和FasL表达的影响。方法：将HepG2 细胞分为LIGHT单独转染组、LIGHT和IFN-γ联合转染组和空白对照组，以脂质体为中介转染HepG2细胞；分别于转染后12、24和48 h收集HepG2细胞，流式细胞术检测转染后HepG2细胞的凋亡率及Fas和FasL的表达。结果：LIGHT基因转染HepG2细胞能明显促进其凋亡，随时间延长凋亡率增加；Fas和FasL在HepG2细胞高表达，以Fas升高程度显著。结论：LIGHT和IFN-γ联合转染HepG2，其凋亡效果优于LIGHT单独转染，主要是通过上调Fas的表达来促进HepG2细胞凋亡。 【关键词】 细胞凋亡·LIGHT·IFN-γ·Fas/Fas-L·转染·HepG2细胞 【ABSTRACT】Objective：To investigate the expression of Fas、FasL and the apoptosis of liver cancer cell line HepG2 transfected with LIGHT and IFN-γ gene mediated by Cationic liposome. Methods：HepG2 cells were divided into two groups (the solo transfection of LIGHT gene and the combined transfection of LIGHT and IFN-γ genes) and the control groups （no transfection）. HepG2 cells were cellected at 12h, 24h and 48h after transfection. The apoptosis of HepG2 cells and the expression of Fas and FasL of the HepG2 cells were investigated with flow cytometry. Results：After transfection, the apoptosis of HepG2 cells increased, and the apoptosis of combined transfection group was higher than the solo transfection of LIGHT（P＜0.01）. The expression of Fas and FasL increased in HepG2 cells（P＜0.01）, but the expression of FasL was lower than that of Fas. Conclusion: The effect of the combined transfection of LIGHT and IFN-γ genes is better than that of the solo transfection of LIGHT gene. LIGHT and IFN-γ can significantly promote cell apoptosis by regulating the expression of Fas. 【KEY WORDS】 Apoptosis·LIGHT·Interferon-γ·Fas/FasL·Transfection·HepG2 cell 　　原发性肝癌居我国恶性肿瘤致死类型的第二位［１］，其中90％以上为肝细胞癌（hepatocellular carcinoma，HCC）。HCC恶性程度高、手术切除率低、预后差，全身化学 治疗 效果不佳，部分原因是HCC细胞的凋亡受阻［２］。本研究将LIGHT和INF-γ配比转染人肝癌细胞HepG2，检测其对凋亡现象及Fas和FasL表达的影响，以探讨促进HCC细胞凋亡的方法。 1　材料与方法 1.1　实验材料与主要试剂　肝癌细胞株HepG2购于南京凯基生物科技有限公司，Fas和FasL抗体购于美国SANTA生物科技有限公司，Mouse IgG1/IgG1、FITC/PE标记抗体购于北京市晶美基因谷科技有限公司，Annexin V/FITC试剂盒购于江苏碧云天生物技术研究所，细胞培养基RPMI-1640和胎牛血清购于美国Gibco公司，Effect