肝癌组织特异性蛋白磷酸酶2A催化性C亚基α亚型显性负性突变体表达载体抑制肝癌移植瘤生长的体内研究  

Effects of the dominant negative form of protein phosphatase 2A catalytic subanit a driven by alpha-feto- protein enhancer/phosphoglycerate kinase promoter on hepatoma cell xenografts

作　　者：龚斐然[1] 李伟[2] 陈凯[2] 陶敏[2] 李道明[2] 徐泽宽[3] 

机构地区：[1]苏州大学附属第一医院血液科,卫生部血栓与止血重点实验室,215006 [2]苏州大学附属第一医院肿瘤科,215006 [3]南京医科大学第一附属医院普外科

出　　处：《中华肝胆外科杂志》2013年第9期696-700,共5页Chinese Journal of Hepatobiliary Surgery

基　　金：国家自然科学基金（81101867、81072031、81200369、81272542）;吴阶平医学基金会临床科研专项资助基金（320.6750.12242）;苏州市科教兴卫青年科技项目（SWKQ1003、SWKQ1011）;苏州市科技计划项目（SYS20J112）;苏州市科技发展计划项目（SYSD2012137）

摘　　要：目的构建腺病毒为载体的肝癌组织特异性启动子驱动的蛋白磷酸酶2A催化性C亚基α亚型显性负性突变体（domina negative form of protein phosphatase 2A catalytic subunitα，DN—PP2Acα）表达载体，研究其对肝癌生长的影响。方法前期研究已构建了甲胎蛋白（alpha-fetoprotein，AFP）基因增强子和磷酸甘油酸激酶（phosphoglycerate kinase，pgk）基因启动子组合形成的肝癌组织特异性AFpg启动子，并将AFpg启动子和DNPP2Aca编码序列构建成AFpg启动子调控的DN—PP2Acα表达载体。将该载体重组到腺病毒载体中。Western印迹法检测PP2Ac表达水平。MTT法检测细胞生长。用荷瘤裸鼠进行体内研究。结果AFpg启动子调控的DN—PP2Acα表达载体可特异性地使DN—PP2Acα表达于AFP阳性肝癌细胞HepG2，并抑制细胞及移植瘤的生长，而对AFP阴性肝癌细胞株SK—HEP1、正常肝细胞株L-02和移植瘤的生长无明显影响。结论AFpg启动子调控的DNPP2Acα表达载体能特异性地抑制AFP阳性肝癌细胞的生长，可用于肝癌组织特异性基因治疗。Objective To investigate the effects of AFP enhancer/pgk promoter driven expression of the dominant negative form of the PP2A catalytic subunit α （DN-PP2Acα） in vivo. Methods The previously constructed AFpg promoter-driven DN-PP2Acα was recombined into an adenovirus, and the expression of PP2Ac was tested using Western blot. Cell growth was tested using the MTT and flat plate clone formation assays. In vivo studies were performed in tumor xenograft models. Resuits AFpg promoter-driven expression of DN-PP2Aα exerted cytotoxic effects against the AFP-pos itive human hepatoma cell line HepG2, but did not affect AFP negative human hepatoma cells （SK- HEP 1） or normal human liver cells （L-02）. Moreover, AFP enhancer/pgk promoter driven expres- sion of DN-PP2Acα inhibited the growth of AFP positive HepG2 tumors in nude mice bearing solid tumor xenografts, but did not affect AFP-negative SK-HEP-1 tumors. Conclusion The recombinant AFP enhancer/pgk promoter driven DN-PP2Aα expression adenovirus presented selective cytotoxicity against AFP-positive hepatoma ceils and provides a useful gene therapy strategy to selectively target hepatocellular carcinoma.

关 键 词：肝癌 甲胎蛋白 磷酸甘油酸酯激酶 蛋白磷酸酶2A 腺病毒 

分 类 号：R735.7[医药卫生—肿瘤]