腺病毒介导MDA-7/IL-24选择性杀伤肝癌 细胞HepG2的研究

目的 ：观察MDA-7/IL-24基因对肝癌的选择性杀伤作用，为肝癌的基因治疗提供理论基础。 方法 ：将携带人MDA-7/IL-24基因的腺病毒Ad.mda-7感染人正常肝细胞L02和肝癌细胞HepG2;用RT-PCR法观察MDA-7/IL-24基因的表达;ELISA方法检测细胞培养上清液中MDA-7/IL-24蛋白的浓度;4甲基偶氮唑蓝染色法（MTT）及Hoechst染色观察MDA-7/IL-24对肝癌细胞的生长抑制和杀伤作用;Annexin-V和PI双染后流式细胞仪检测2种细胞的凋亡;用流式细胞仪检测细胞周期。 结果 ：复制缺陷型腺病毒能介导外源基因MDA-7/IL-24在肝癌细胞株HepG2和正常细胞L02中的高效表达;细胞培养上清液中有MDA-7/IL-24蛋白的表达; MDA-7/IL-24能明显抑制肝癌细胞生长并可促进肝癌细胞的凋亡;MDA-7/IL-24阻滞肝癌细胞于G2/M期，能选择性杀伤肝癌细胞而对正常的肝细胞无阻滞作用和毒性作用。结论 ：复制缺陷型重组腺病毒载体Ad.mda-7能介导MDA-7/IL-24基因在人肝癌细胞中高效表达，促使细胞增殖阻滞及诱导肿瘤细胞凋亡，选择性地杀伤肝癌细胞HepG2，而对正常肝细胞L02无任何毒性作用。

Adenovirus mediated MDA-7/IL-24 gene transfer selectively killshepatocellular carcinoma lines HepG2

Objective To investigate the selective killing effect of MDA/IL-24 on human hepatocellular carcinoma line HepG2 in vitro and provide a theoretical basis for gcne therapy of hepatocellular carcinoma. Methods The MDA-7/IL-24 gene was transfected into human hepatocullulat carcinoma cell line HepG2 and normal liver cell llne L02 with a replication-incompetent adenovirus vector. The mRNA and protein expression of MDA7/IL-24 in HepG2 and L02 cells was examined by RT-PCR and ELISA assay respectively. MTT assay and flow cytometry were used to study tumor cell proliferation and cell cycle in vitro. Hoechst and Annexin-V and PI staining were studied to indicate the apoptosis. Results RT-PCR confirmed that the exogenous MDA-7/IL-24 gene was expressed in HepG2 and L02 cells. The protein product was confirmed by assay of the supernatant with ELISA. MTT and apoptosis test indicated MDA-7/IL-24 induced growth suppression and cell apoptosis of the HepG2 cell in vitro but not in cell line L02 , and cell cycle test revealed MDA-7/IL-24 could block HepG2 cell in G2/M hut not in L02. Conclusions MDA-7/IL-24 selectively induces growth suppression and apoptosis in lines HepG2 in vitro but not in L02 cell, which indicates that adenovirus mediated MDA-7/IL-24 can be an excellent tool for gene therapy in hepatocellular carcinoma.