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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 killing effect of E1B5 5kDa-deficient adenovirus dl152 on liver cancer cells. Methods Human hepatoma cells with different genotypes of p53 were infected with dl152o respectively. The viable cells were detected by staining on the 4th day after infection. RT PCR was used to detect the changes of p5 3 and p2 1Waf 1 gene expression in the cells. Adenovirus infection was confirmed by detecting the expression of the adenoviral capsid protein hexo gene. The dl152 was injected intratumorally into SCID nude mice to observe the in vivo killing effect of dl152 on liver cancer cells. RESULTS: p53-deleted hepatocellular carcinoma cell Hep3B was most sensitive to dl152-induced cytotoxicity, with more than 60% of cells being killed, and less than 20% of PLC/PRF/5 (p53 mutant) and HepG2 (p53 gene is wild-type) liver cancer cells were killed. After adenovirus infection, the expression levels of p53 and p2 1Waf1 genes in HepG2 cells were significantly increased. Intratumoral injection of dl152, can significantly inhibit the growth of transplanted tumors in Hep3B nude mice, but there was no significant growth inhibitory effect on PLC/PRF/5 and HepG2 nude mice. Conclusion E1B5 5kDa-deficient adenovirus can selectively kill p53-deleted hepatoma cells, which is a potential therapeutic method for tumors.