反义HIF-1α基因治疗人肝癌裸鼠移植瘤的实验研究

摘要：目的:探讨反义HIF-1α基因对人肝癌裸鼠皮下移植瘤的影响及其相关机制。方法:使用人原发性肝癌细胞株SMMC-7721建立人肝癌裸鼠皮下移植瘤动物模型。待肿瘤生长至直径约0.4cm时，将荷瘤裸鼠随机分为3组，分别注射生理盐水、质粒PcDNA3和HIF-1α/PcDNA3B，质粒用脂质体DOTAP介导转染细胞。观察各组动物的肿瘤生长曲线；取肿瘤标本作免疫组织化学检查（SABC法）及蛋白质印迹检查，检测各组肿瘤的VEGF和HIF-1α表达及微血管密度（MVD）和细胞凋亡。结果:HIF-1α/PcDNA3B治疗组各时点的肿瘤体积，以及肿瘤组织中HIF-1α蛋白、MVD，VEGF表达均低于对照组，而细胞凋亡指数高于对照组，差异均有显著性（P＜0.05）。结论:通过阻断癌细胞的缺氧适应途径，反义HIF-1α基因治疗肝癌有抑制肿瘤生长、抑制肿瘤血管生成及诱导细胞凋亡的作用。

Experimental study of treating implanted human primary hepatic carcinoma of nude mice by antisense HIF-1α gene in-vivo

Abstract:Objective:To observe the effects and mechanism of antisense HIF-1α gene on implanted human primary hepatic carcinoma of nude mouse．Methods:Tumors were established by subcutaneous injection of 3×106 tumor cells，named SMMC-7721，into the back of nude mice．Once the tumor grew to 0.4cm in diameter, the mice were divided randomly into three groups with injected respectively with NS、empty plamid PcDNA3、and antisense HIF-1α plasmid．Half group of the nude mice were used to observe the tumor growth curve，and the other half were to obtain the tumor samples to be used in the examinations of the expression of angiostatin、HIF-1α, VEGF, MVD and cell apoptosis．Results:Tumors grew rapidly in the control group than that in the antisense HIF-1α group. In antisense HIF-1α gene therapy resulted in low expression of HIF-1α, MVD and VEGF than those in 2 control groups(P<0.05~0.01); but in HIF-1α group. AI was higher than that in the control group（P＜0.01）.Conclusions:Antisense HIF-1α gene therapy is a promising approach to treat human liver cancer by suppression of tumor growth and tumor angiogenesis and by induction of cell apoptosis．