腺病毒介导的p53抑制肝癌生长的实验研究

目的探讨外源性野生型Ｐ５３（Ｗｉｌｄ－ｔｙｐｅ ｐ５３， ＷＴ－ｐ５３）基因对肝癌细胞生长的影响。方法通过表达ＷＴ－ｐ５３的重组腺病毒载体（Ａｄ－ｐ５３），将ｐ５３基因导入肝癌细胞株中，用ＭＴＴ法检测Ａｄ－ｐ５３对各细胞株生长特性的影响，流式细胞仪分析各组细胞中ＤＮＡ含量及凋亡的比例。建立裸小鼠移植瘤模型后，瘤内注射Ａｄ－ｐ５３。最后处死动物，移植瘤称重， ｗｅｓｔｅｒｎ　ｂｌｏｔ检测ｐ５３和ｐ２１蛋白的表达情况。结果外源性ｐ５３基因的导入可导致肝癌细胞生长的明显抑制，表现为既诱导细胞凋亡，又诱导细胞周期阻滞。瘤内注射Ａｄ－ｐ５３后移植瘤生长明显受抑制，并出现瘤组织中ｐ５３和ｐ２１蛋白表达的上调。结论腺病毒介导的ｐ５３基因治疗能有效控制肝癌的生长。

Therapeutic potential of recombinant adenovirus expressing p53 in hepatocellular carcinoma cell lines

Objective To investigate the effects of the recombinant adenoviral vector Ad-p53 on the biological behavior of hepatocellular carcinoma (HCC) cells in vitro and in vivo. Methods With recombinant adenoviral vector expressing WT-p53 (Ad-p53), p53 gene was transfected into the HCC cell line, PLC/PRF/5.The cytotoxicities of Ad-p53 to cells were measured by MTT assay. Cell growth properties and cell cycle patterns were assessed with flow cytometry. The animal model was developed by injecting HCC cells into the dorsum of nude mice. Ad-p53 was injected intratumorally. The animals were killed, and then excised tumors were weighed and analyzed for p53 and p21 protein expression using western blot assay. Results The introduction of exogenous wild-type p53 resulted in the inhibition of cell growth, high G2/M ratio and cell apoptosis, and low S ratio in PLC/PRF/5. The expression of both p53 and p21 proteins was upregulated in the cells. Conclusions Replication-deficient adenoviral vector expressing WT-p53 may be useful for gene therapy of HCC.