靶向性重组腺病毒对裸鼠肝癌种植瘤的治疗作用  

Therapeutic Effect of Recombinant Adenovirus Containing Ad-EAFP-PALB/r-Caspase-3 on Primary Hepatocellular Carcinoma

作　　者：王炜[1] 沈浩元[1] 刘猛[1] 

机构地区：[1]广州医学院第一附属医院肝胆外科,广东广州510120

出　　处：《中国普外基础与临床杂志》2011年第7期722-727,共6页Chinese Journal of Bases and Clinics In General Surgery

基　　金：国家自然科学基金(项目编号:30571829);广东省医学科研基金(项目编号:A2006282)~~

摘　　要：目的观察靶向性重组腺病毒Ad-EAFP-PALB/r-Caspase-3对原发性肝癌HepG2细胞及裸鼠皮下种植瘤的治疗作用。方法采用先前构建的靶向性重组腺病毒Ad-EAFP-PALB/r-Caspase-3感染肝癌HepG2细胞、乳腺癌MDA-MB-231细胞以及正常肝L-02细胞,观察细胞形态学改变,并通过流式细胞术检测细胞凋亡指数;采用裸鼠皮下种植瘤模型、重组腺病毒瘤内给药治疗,观察肿瘤生长情况及病理学改变。结果 流式细胞学可见HepG2细胞凋亡和明显的凋亡峰,MDA-MB-231和L-02细胞凋亡指数分别为0和7.3%,明显低于HepG2细胞(48.2%),P<0.01。种植肿瘤细胞2周后HepG2细胞组、MDA-MB-231细胞组和对照组肿瘤体积大小分别为(0.26±0.31)cm3、(0.25±0.15)cm3和(0.26±0.28)cm3,各组间差异无统计学意义(P>0.05);种植肿瘤细胞4周后HepG2细胞组、MDA-MB-231细胞组和对照组肿瘤体积大小分别为(0.53±0.12)cm3、(0.49±0.22)cm3和(0.54±0.13)cm3,各组间差异无统计学意义(P>0.05);HepG2细胞组肿瘤经重组腺病毒Ad-EAFP-PALB/r-Caspase-3治疗后8周时肿瘤体积为(0.65±0.13)cm3,明显小于MDA-MB-231细胞组〔(1.27±0.32)cm3〕和对照组〔(1.43±1.09)cm3〕,P<0.01,而后两组之间比较差异无统计学意义(P>0.05)。治疗后HepG2细胞组肿瘤细胞出现坏死区,伴淋巴浸润细胞和凋亡细胞;对照组肿瘤细胞区域内未见明显坏死区;而MDA-MB-231细胞组肿瘤细胞呈实性排列,呈小腺腔样结构,异型性明显,瘤巨细胞及多核瘤细胞可见,治疗前、后未见明显变化。结论 靶向性重组腺病毒Ad-EAFP-PALB/r-Caspase-3具有有效靶向性凋亡诱导作用,体内、外治疗原发性肝癌有效,可能为原发性肝癌的靶向性基因治疗提供新途径。Objective To evaluate therapeutic effect of the targeting recombinant adenovirus Ad-EAFP-PALB/r-Caspase-3 on primary hepatocellular carcinoma cells（HepG2） and subcutaneous implanted tumors of nude mice.Methods HepG2 cells,breast cancer MDA-MB-231 cells,and normal hepatic L-02 cells were infected with the previously constructed targeting recombinant adenovirus Ad-EAFP-PALB/r-Caspase-3,then morphological change was observed and apoptosis index was detected by cell morphologic assay.Tumor growth and pathological changes were observed after the implanted tumors model of nude mice were injected with the recombinant adenovirus.Results Both apoptosis and obvious apoptotic peak of cells were observed by flow cytometry（FCM） in HepG2 cell group.The apoptosis index of MDA-MB-231 and L-02 cells was 0 and 7.3%,respectively,which were significantly lower than that of HepG2 cells（48.2%）,P〉0.01.The volume of subcutaneous tumor of nude mice was（0.26±0.31） cm^3 in HepG2 cell group,（0.25±0.15） cm^3 in MDA-MB-231 cell group,and（0.26±0.28） cm^3 in control group on two weeks after implantation of cancer cells,and no statistical difference was found among them（P〉0.05）.The volume of subcutaneous tumor of nude mice was（0.53±0.12） cm^3 in HepG2 cell group,（0.49±0.22） cm^3 in MDA-MB-231 cell group,and（0.54±0.13） cm^3 in control group on four weeks after implantation of cancer cells,and the difference was not significant among them（P〉0.05）.But on eight weeks after implantation of cancer cells,the volume of tumor was（0.65±0.13） cm^3 in HepG2 cell group,（1.27±0.32） cm^3 in MDA-MB-231 cell group,and（1.43±1.09） cm^3 in control group,which suggested that the growth of tumor in HepG2 cell group slowed down significantly when compared with MDA-MB-231 cell group and control group（P〉0.01）,although the difference in the latter two groups was not significant（P〉0.05）.The necrosis regions were found with lymphocytic infiltration and apoptosis cells in HepG2 cell group aft

关 键 词：肝细胞性肝癌 CASPASE-3 凋亡 种植瘤 裸鼠 

分 类 号：R735.7[医药卫生—肿瘤]