Ad-ING4抑制裸鼠人肝癌移植瘤生长的体内实验研究

The growth-inhibition of hepatocarcinoma xenografts by adenovirus-mediated ING4 gene in nude mice

目的研究携带ING4基因的重组腺病毒(Ad-ING4)对人肝癌细胞株SMMC-7721移植瘤生长的抑制作用及其潜在的分子机制。方法将扩增的Ad-ING4感染SMMC-7721细胞,Western blot法检测Ad-ING4在SMMC-7721细胞中的转录。用SMMC-7721细胞建立裸鼠人肝癌移植瘤模型,予Ad-ING4(1×109pfu/ml)50μl对移植瘤瘤体注射治疗,观察肿瘤生长变化;3周后处死裸鼠,摘除瘤体,称瘤重;免疫组化法检测p21、COX-2、Fas、Bcl-2、Bax、Caspase-3、VEGF、CD34等因子的表达。结果 Ad-ING4在SMMC-7721细胞中成功表达;Ad-ING4对裸鼠人肝癌移植瘤有明显的生长抑制作用,Ad-ING4组瘤体显著减小(P<0.01),抑瘤率达41.64%;免疫组化检测显示,Ad-ING4抑癌的分子机制可能与上调p21、Fas、Bax和Caspase-3的表达及下调COX-2、Bcl-2、VEGF和CD34的表达有关。结论 Ad-ING4能有效抑制SMMC-7721裸鼠人肝癌移植瘤的生长,其机制可能通过抑制肿瘤细胞生长、血管形成和激活细胞凋亡等多个途径共同发挥抑癌效应。

Objective To study the inhibitory effects and anti-cancer mechanism of adenovirus-mediated ING4 gene（Ad-ING4） on SMMC-7721 hepatocarcinoma xenografts in nude mice.Methods After transfection of purified Ad-ING4 into SMMC-7721 hepatocarcinoma cells,the expression of Ad-ING4 was detected by Western blot.Nude mice of the subcutaneous tumor models were established with SMMC-7721 cells.Athymic nude mice bearing SMMC-7721 tumors were intratumorally injected with Ad-ING4（50μl,1×10^9pfu/ml）.Tumor growth was recorded,and all nude mice were killed at the end of the experiment to observe the growth of xenografts.The expressions of p21,COX-2,Fas,Bcl-2,Bax,Caspase-3,VEGF and CD34 in tumor tissues were detected by immunohistochemistry.Results Ad-ING4 was successfully expressed in SMMC-7721 cells.Tumor growth was significantly inhibited in the Ad-ING4 group as compared with that in the Ad group and the PBS group（P〈0.01）.The weight inhibitory rate was 41.64% in the Ad-ING4 group.Immumohistochemistry displayed that the expressions of p21,Fas,Bax and Caspase-3 were up-regulated,and the expressions of COX-2,Bcl-2,VEGF and CD34 were down-regulated in the Ad-ING4 group.Conclusion Ad-ING4 could inhibit the growth of SMMC-7721 hepatocarcinoma xenografts in nude mice,may be via inhibiting cell growth,activating the apoptosis pathway and inhibiting tumor angiogenesis.