黑色素瘤分化相关基因-7和白介素-24选择性杀伤肝癌细胞和诱导增殖阻滞的体外研究  

Melanoma differentiation associated gene-7 and interleukin-24 selectively induces growth arrest and apoptosis in hepatocellular carcinoma cell lines in vitro

作　　者：王从俊[1] 彭志海[1] 于愿[2] 陈堃[2] 郑建伟[2] 胡回忆[2] 吉文伟[2] 薛新波[2] 

机构地区：[1]上海交通大学附属上海市第一人民医院普通外科,200080 [2]华中科技大学同济医学院附属同济医院胆胰外科中心

出　　处：《中华外科杂志》2007年第17期1202-1205,共4页Chinese Journal of Surgery

基　　金：湖北省科技攻关重点资助项目(2006AA301B52-4)

摘　　要：目的探讨黑色素瘤分化相关基因-7/白介素-24基因(mda-7/IL-24)对不同种类肝癌细胞的选择性杀伤作用。方法将携带人 mda-7/IL-24基因的复制缺陷型腺病毒(Ad.mda-7)感染人正常肝细胞和各种肝癌细胞,通过逆转录聚合酶链反应(RT-PCR)和酶联免疫吸附(ELISA)实验观察mda-7/IL-24基因的表达,MTT 法观察肝癌细胞的生长抑制,Hoechst 染色及 Annexin-V 和 PI 双染后流式细胞仪检测细胞的凋亡情况及用流式细胞仪检测细胞周期。结果 RT-PCR 和 ELISA 提示Ad.mda-7能介导外源基因 mda-7/IL-24在各种肝癌细胞株和正常肝细胞中高效表达。MTT 实验结果提示 mda-7/IL-24能明显抑制各种肝癌细胞的生长,Hoechst 染色和流式细胞仪检测提示 mda-7/IL-24能选择性杀伤肝癌细胞,细胞周期分析提示 mda-7/IL-24阻滞肝癌细胞在 G2/M 期,同时对正常的肝细胞没有促凋亡和增殖阻滞作用。结论 mda-7/IL-24基因能选择性杀伤各种肝癌细胞,促进细胞增殖阻滞及诱导细胞凋亡。Objective To investigate the effect of melanoma differentiation associated gene-7/ interleukin-24 （mda-7/IL-24） on the hepatocullular carcinoma cell lines and normal liver cell line in vitro. Methods Hepatocellular carcinoma cell lines HepG2, SMMC7721, Hep3B, MHCC97L, M6 and normal liver cell line L02 were infected with Ad. mda-7. The gene expression of mda-7/IL-24 in these cell lines was confirmed by RT-PCR and ELISA assay. MTY assay and flow cytometry were used to study tumor cell proliferation and cell cycle in vitro. Hoechst staining and cytometry assay after Annexin-V and PI staining were studied to indicate the apoptosis effect. Results It was confirmed by RT-PCR that the exogenous mda- 7/IL-24 gene expressed in all of these cells. The mda-7/IL-24 protein product was confirmed by assaying the supernatant with ELISA. MTT and apoptosis test indicated mda-7/IL-24 can induce the hepatocellular carcinoma cell lines growth suppression,apoptosis in vitro but not in normal liver cell line L02, cell cycle test revealed mda-7/IL-24 can block cancer cell lines in G2/M but not in L02. Conclusions mda-7/IL-24 selectively induces growth suppression, apoptosis in hepatocellular carcinoma lines but not in normal liver cell in vitro.

关 键 词：癌 肝细胞 基因治疗 凋亡 增殖阻滞 

分 类 号：R735.7[医药卫生—肿瘤]