血管内皮生长因子受体启动子双自杀基因治疗乳腺癌  

In vivo anti-tumor effect of the recombinant adenoviruses containing CD/TK double suicide gene driven by KDR promoter on breast cancer

作　　者：苏国强[1] 苏刚[2] 黄宗海[3] 

机构地区：[1]厦门大学附属第一医院普通外科,361003 [2]郑州大学第一附属医院心外科 [3]南方医科大学附属珠江医院普通外科

出　　处：《中华实验外科杂志》2010年第11期1666-1668,F0003,共4页Chinese Journal of Experimental Surgery

基　　金：国家“863”计划资助项目（2001AA217171）;厦门市卫生局资助项目（WZK0609）

摘　　要：目的 探讨腺病毒介导的KDRP-CD/TK融合基因对乳腺癌裸鼠体内抑瘤作用的特点.方法 以MCF-7细胞株建立裸小鼠乳腺癌动物模型,随机分为Ⅰ组[注射重组腺病毒AdKDRP-CDglyTK与前药5-氟胞嘧啶（5-FC）与丙氧鸟苷（GCV）]、Ⅱ组（仅注射前药5-FC与GCV）、Ⅲ组（仅注射重组腺病毒AdKDRP-CDglyTK）及Ⅳ组（空白对照,不施加任何处理）.治疗结束后,计算抑瘤率、常规病理检查、逆转录-聚合酶链反应（RT-PCR）检测瘤体CD/TK融合基因的表达以及微血管密度（MVD）变化的检测,同时观察该治疗体系有无系统毒性.结果 各组瘤重如下：Ⅰ组（实验组）（25.04±3.09）mg、Ⅱ组（498.07±4.47）mg、Ⅲ组（501.94±4.50）mg、Ⅳ组（503.79±6.27）mg.可见Ⅰ组肿瘤逐渐缩小,余各组肿瘤逐渐增大（F=12 727.420,P＜0.01） 第Ⅱ、Ⅲ、Ⅳ组肿瘤生长差异无统计学意义（P＞0.05） Ⅰ组有CD/TK基因表达且MVD（1.05±0.04）较对照组（4.15±0.10）变小（t=64.126,P＜0.01） 常规病理检查发现Ⅰ组组织片状坏死,且该治疗体系对重要脏器无毒性作用.结论 KDR启动子驱动双自杀基因体系对裸鼠皮下移植瘤有明显的生长抑制作用,其机制涉及对肿瘤及其血管内皮细胞的双重杀伤.Objective To examine anti-tumor effect of the recombinant adenoviruses containing Herpes simplex virus thymidine kinase/E. coli cytosine deaminase （CD/TK） double suicide gene driven by kinase insert domain-containing receptor （KDR） promoter on breast tumor in vivo. Methods Four- to 6-week-old female BALB/C nu/nu euthymic mice were used as hosts for MCF-7 cell xenografts, and MCF-7 cells were injected subcutaneously into the mammary fat pad. The rats were randomly divided into four groups according to the treatment regimen （ group Ⅰ： viruses ＋ 5-FC ＋ GCV, group Ⅱ： viruses only,group Ⅲ ： 5-FC ＋ GCV only, group Ⅳ： blank）. Tumor growth inhibition rate was calculated at the end of treatment. And HE staining of tumor tissues was performed for histological examination and immunohistochemistry method was used to detect the microvessel density （MVD） of tumor tissues. CDglyTK gene expression of tumor tissues was detected by reverse transcription-polymerase chain reaction （RT-PCR）. Systemic toxicity of recombinant adenoviruses and prodrugs was determined. Results Weight of tumor in groups Ⅰ, Ⅱ, ⅢI and Ⅳ was （25.04±3.09）, （498.07±4.47）, （501.94±4.50） and （503.79±6. 27） mg respectively. Tumors were decreased in size during treatment of double suicide gene system （ group Ⅰ）, but increased in size in the other sgroups （ F = 12 727. 420, P 〈 0. 01 ）. Moreover, tumor weights had no significant difference among groups Ⅱ , Ⅲ and Ⅳ （P 〉 0.05）. HE staining showed lamellar necrosis of tumor cells and the MVD was reduced in tumors treated with the recombinant adenoviruses and prodrugs, but not in control groups （t = 64. 126,P 〈 0.01 ）. The CDglyTK gene expression was confirmed by RT-PCR in tumor in nude mice treated with AdKDRP-CDglyTK. There were no histological changes in the heart, liver, lung, kidney and small intestine in in treated groups. Conclusion CDglyTK gene system driven by KDR promoter can significantly inhibit the tumor gro

关 键 词：乳腺肿瘤 微血管密度 基因 自杀 腺病毒 

分 类 号：R737.9[医药卫生—肿瘤]