抑癌基因VHL治疗实体肿瘤的实验研究  

Gene transfer of von Hippel-Lindau inhibits the growth of transplanted solid tumors

作　　者：孙学英[1] 王建立[2] 唐波[1] 刘凤军[2] 乔海泉[1] 姜洪池[1] 

机构地区：[1]哈尔滨医科大学第一临床医学院普通外科黑龙江省肝脾外科中心,150001 [2]山东大学齐鲁医院普通外科

出　　处：《中华胃肠外科杂志》2005年第3期241-244,共4页Chinese Journal of Gastrointestinal Surgery

基　　金：国家自然科学基金资助项目(30471681;30100178)

摘　　要：目的探讨抑癌基因VHL治疗EL-4实体肿瘤的效果。方法皮下注射EL-4淋巴瘤细胞建立C57BL/6小鼠实体肿瘤模型。当肿瘤生长至直径分别为0.1cm和0.4cm时,将小鼠随机分为治疗组和对照组,每组各6只。治疗组瘤体内采用脂质体法注射pcDNA3-VHL基因,对照组注射空载体,观察肿瘤生长情况。用免疫组织化学(免疫组化)和(或)Western印迹方法检测VHL基因、血管内皮生长因子(VEGF)、缺氧诱导因子-1α(HIF-1α)、抗凋亡因子(bcl-2)在瘤组织内的表达,测定肿瘤的微血管密度(MVD);TUNEL法测定肿瘤的凋亡指数(AI)。结果VHL基因治疗能够根除小的肿瘤(直径0.1cm),但只能减慢而不能消除较大肿瘤(直径0.4cm)的生长。治疗组VHL基因在瘤组织内高表达,VEGF、HIF-1α和Bcl-2的表达显著低于对照组,MVD明显低于对照组(P<0.05),AI显著高于对照组(P<0.01)。结论VHL基因疗法对小鼠EL-4实体肿瘤的生长有显著的抑制作用。Objective To explore the effect of von Hippel Lindau(VHL) gene on growth of EL 4 solid tumors in vivo. Methods C57BL/6 mice model of solid tumor s was established by subcutaneous injection of EL 4 lymphoma cells. Mice were r andomly divided into two groups as treatment group(n=6) and control group(n=6) w hen tumor diameter increased to 0.1 cm and 0.4 cm respectively. Plasmid pcDNA3 VHL was injected into solid tumor in treatment group,empty pcDNA3 vector in cont rol group. The growth of tumor was observed. Immunohistochemistry and Western bl ot analysis were used to examine the transgenic expression of VHL,hypoxia induci ble factor 1α(HIF) 1α,bcl 2 and VEGF. Microvessal density(MVD) and apoptosi s index(AI) of tumors were also detected. Results VHL gene transfer eradicated t umors with small size(0.1 cm diameter),but it only retarded the growth of larg e tumors(0.4 cm diameter). VHL was overexpressed,the expression levels of VEGF,H IF 1αand bcl 2 were reduced in treatment group compared with those in the con trol group. The level of MVD was significantly lower in treatment group(P< 0.05) ,but AI was higher in treatment group compared with those in the control group(P < 0.01). Conclusion VHL gene therapy can inhibit the growth of EL 4 solid tumor in vivo.

关 键 词：实体肿瘤 抑癌基因 血管内皮生长因子(VEGF) 实验研究 微血管密度(MVD) C57BL/6小鼠 缺氧诱导因子-1α Westem印迹 HIF-1α VHL基因 EL-4 免疫组织化学 TUNEL法 肿瘤生长 淋巴瘤细胞 对照组 Bcl-2 治疗组 肿瘤模型 皮下注射 

分 类 号：R730.5[医药卫生—肿瘤]