E1B55kDa缺陷型腺病毒对人肝癌细胞的杀伤研究

Anti-tumor effect of E1B55kDa-deleted adenovirus on human hepatocellular carcinoma cell lines

目的 探讨E1B5 5kDa缺陷型腺病毒dl15 2 0对肝癌细胞的体内外杀伤作用。方法 用dl15 2 0分别感染p5 3基因型不同的人肝癌细胞 ,感染后第 4天用染色的方法检测存活细胞。用RT PCR检测细胞内p5 3和p2 1Waf 1基因表达的改变 ,通过检测腺病毒衣壳蛋白hexon基因的表达证实腺病毒的感染。在SCID裸鼠瘤体内注射dl15 2 0 ,观察dl15 2 0对肝癌细胞的体内杀伤作用。结果 p5 3基因缺失的肝癌细胞Hep3B对dl15 2 0诱导的细胞毒性作用最敏感 ,超过 6 0 %的细胞被杀伤 ,而不足2 0 %的PLC/PRF/ 5 (p5 3基因突变型 )和HepG2 (p5 3基因野生型 )肝癌细胞被杀伤。腺病毒感染后 ,HepG2细胞内p5 3和p2 1Waf 1基因表达水平均明显升高。瘤体内注射dl15 2 0 ,可显著抑制Hep3B裸鼠移植瘤的生长 ,而对PLC/PRF/ 5和HepG2的裸鼠移植瘤则无明显的生长抑制作用。结论 E1B5 5kDa缺失的腺病毒可以选择性地杀伤p5 3基因缺失的肝癌细胞 ,是一种潜在的肿瘤治疗手段。

Objective To investigate the in vitro and in vivo anti tumoral effect of E1B55kDa deleted adenovirus (dl1520) on human hepatocellular carcinoma (HCC) cell lines.Methods Human HCC cell lines with different p53 genotypes were infected with dl1520. Four days after infection, the percentage of survival cells was determined by colorimetric assay. RT polymerase chain reaction (PCR) was used to examine alterations of p53 and p21 Waf 1 expression. Adenovirus hexon gene expression was applied to prove the presence of adenovirus infections. Effects on vivo anti tumor dl1520 were examined with intra tumor injection of dl1520 in the SCID mice.Results After the infection, the p53 null HCC cell line, Hep3B was most vulnerable to dl1520 induced cytotoxic effect with a cell death rate of more than 60%. Whereas, less than 20% PLC/PRF/5 (p53 mutant) and HepG2 (p53 wild type) HCC cell lines were killed. The expression of p53 and p21 Waf 1 was considerably enhanced after the dl1520 infection in HepG2. The intra tumor injection of dl1520 significantly reduced the tumor growth in Hep3B xenografts. In contrast, no obvious tumor repression was observed in HepG2 and PLC/PRF/5 xenografts.Conclusion dl1520 selectively kills the p53 null hepatocellular carcinoma, showing potential application for cancer treatment. [