摘要
目的利用携带人MDA-7/IL-24基因的复制缺陷型腺病毒(Ad.mda7)作为载体,感染不同转移潜能的肝癌细胞MHCC97H,MHCC97L,Hep3B和正常的肝细胞L02,观察该基因对肝癌的选择性杀伤作用和增殖抑制作用。方法将携带人MDA-7/IL-24基因的腺病毒Ad.mda-7感染正常肝细胞L02和肝癌细胞MHCC97H,MHCC97L和Hep3B,通过RT-PCR和ELISA方法观察MDA7基因的表达,通过四甲基偶氮哩蓝染色法(MTT)观察该基因对肝癌细胞的生长抑制,Hoechst染色和流式细胞仪观察MDA-7对肝癌细胞的杀伤作用,流式细胞仪检测细胞周期变化。结果RT-PCR提示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MHCC97H,MHCC97L,Hep3B和正常细胞L02中高效表达;ELISA方法检测提示细胞培养上清液中MDA-7/IL-24蛋白的表达,而且随着感染时间延长表达量升高;MTT实验结果表明MDA7能明显抑制肝癌细胞的生长,Hoechst染色和流式细胞仪提示MDA7能促进肝癌细的凋亡;细胞周期检测提示肝癌细胞大量被阻滞在G2/M期,正常细胞没有增殖阻滞作用。各项结果提示MDA7对正常的肝细胞没有促进凋亡和增殖阻滞的作用。结论Ad.mda-7选择性的杀伤肝癌细胞,促进细胞增殖阻滞及诱导肿瘤细胞凋亡与肝癌细胞的转移潜能无关。
Objective To investigate the effects of MDA/IL-24 on hepatocellular carcinoma (HCC) cell lines MHCC97H, MHCC97L and Hep3B with different metastatic potentials and normal liver cell line L02 in vitro. Methods The 3 HCC cell lines and normal L02 were infected by Ad. mda- 7 and the mRNA expression of MDA 7/IL-24 in these cell lines was determined by RT-PCR. Protein expression was confirmed by ELISA. MTT assay and flow cytometry were used to study tumor cell proliferation and cell cycle in vitro. Heochst and Annexin-V and PI staining were performed to indicate the apoptotic effect. Results It was confirmed by RTPCR and ELISA that the exogenous MDA-7/ IL-24 gene expressed the 3 HCC cell lines and normal L02. MTT and cycle test showed that Ad. mda- 7 could induce HCC cell line growth suppression and block cancer cells in G2/M but not in L02. Hoechst staining and flow cytometry revealed that MDA-7/IL-24 could induce apoptosis in HCC cell lines but not in normal liver cells. Conclusions MDA-7/IL-24 can selectively induce growth suppression and apoptosis in HCC cell lines and this induction is independent of different metastatic potentials.
出处
《中华肝胆外科杂志》
CAS
CSCD
2007年第10期701-704,共4页
Chinese Journal of Hepatobiliary Surgery
基金
湖北省科技攻关重点项目(2006AA304B05)致谢:美国哥伦比亚大学Fisher教授在关键技术上的指导,感谢同济医院儿科肾病研究室周建华教授对该研究的技术指导及实验设备的支持.