Ａｄ-ＩＮＧ４抑制裸鼠人肝癌移植瘤生长的体内实验研究

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，Ｗｅｓｔｅｒｎｂｌｏｔ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（１×１０^９ｐｆｕ／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。

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

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.