内皮抑制素基因治疗小鼠肝癌及其机制研究  

The Research of Endostatin Gene Therapy on Mouse Hepatoma and the Mechanism

作　　者：黎培员[1] 林菊生[1] 冯作化[2] 张桂梅[2] 沈霞[1] 谢娜[1] 田德安[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院肝病研究所,武汉430030 [2]华中科技大学同济医学院生物化学与分子生物学系

出　　处：《肿瘤防治研究》2004年第11期661-663,F002,共4页Cancer Research on Prevention and Treatment

基　　金：国家重点基础研究发展(973)计划资助项目(2002CB513100)。

摘　　要：目的探讨内皮抑制素质粒重复注射治疗小鼠肝癌的效果及其作用机制。方法内皮抑制素质粒转染BHK21,ELISA检测培养上清中内皮抑制素的含量,并用此上清作用于ECV304内皮细胞,MTT法检测内皮细胞的增殖。小鼠股部肌内接种H22细胞,反复注射内皮抑制素质粒/PVP,观察肿瘤的形成。ELISA检测血清中内皮抑制素的含量,免疫组化观察肿瘤血管密度,TUNEL检测肿瘤细胞的凋亡。结果转染上清中可以检测到分泌出的内皮抑制素,并可抑制内皮细胞的增殖,抑制率约29.2%。治疗组血清内皮抑制素水平升高,瘤内血管减少,肿瘤细胞凋亡增加,肝癌生长受抑,抑制率为56.4%。结论PVP介导的内皮抑制素基因治疗可以抑制小鼠肝癌血管形成,从而较好地抑制了肝癌的生长。Objective To study the inhibitory effect of endostatin gene therapy on mouse hepatoma and the mechanism of which.Methods BHK 21 was transfected with pSecES, in the supernant of which endostatin was assayed by ELISA and the supernant was used to culture endothelium cells ECV304. Proliferation of the latter was evaluated by MTT. Then H22 cells were inoculated into the leg muscle of mouse, which was injected with pSecES/PVP repeatedly. Tumor weight and serum endostatin were assayed. Tumor microvessel density and apoptosis of tumor cells were also displayed by immumohistochemistry and TUNEL.Results Endostatin can be secreted and can inhibit the proliferation of ECV304 by 29.2% . Tumor growth was inhibited by 56.4% companied with higher serum endostatin, fewer tumor vessels and more apoptosis cells.Conclusion Mouse H22 hepatoma can be inhibited by repeated endostatin gene therapy formulated with PVP due to decreased vessel density.

关 键 词：肝癌 内皮抑制素 基因治疗 

分 类 号：R730.5[医药卫生—肿瘤] R735.7[医药卫生—临床医学]