摘要
目的探讨重组腺病毒Ad-VT对人原发性肝癌HepG-2细胞的体内外抑制作用。方法使用重组腺病毒Ad-VT、Ad-T、Ad-vp3以及Ad-mock感染HepG-2细胞,通过MTS检测重组腺病毒对HepG-2细胞的杀伤抑制作用;通过Hoechst染色、Annexin V-FITC/PI以及JC-1染色对重组腺病毒抑制HepG-2细胞的主要途径进行分析;通过构建体内裸鼠荷瘤模型,分析重组腺病毒对HepG-2细胞的体内抑瘤作用。结果Ad-VT可抑制HepG-2细胞的增殖,并呈时效和剂效关系,在100 MOI的剂量下,72 h时抑制率达最高值64.22%,与对照病毒Ad-T的50.52%和Ad-vp3的43.17%比较差异有统计学意义(P<0.05)。Ad-VT通过内源性凋亡途径引起HepG-2细胞的死亡。体内抑瘤试验中Ad-VT能够抑制肿瘤的生长,且能够延长小鼠的存活时间并提高存活率。结论Ad-VT能够诱导肝癌细胞凋亡,改变肝癌细胞线粒体膜电位,有效地抑制肝癌细胞增殖和肿瘤生长。
Objective To investigate the anti-tumor action of the recombinant adenovirus Ad-VT on human primary liver cancer HepG-2 cells in vitro and in vivo.Methods HepG-2 cells were infected with the recombinant adenoviruses Ad-VT,Ad-T,Ad-vp3,and Ad-mock.The inhibitory effect of the recombinant adenoviruses on HepG-2 cells was determined using MTS.The main pathways by which recombinant adenoviruses inhibited HepG-2 cells were analyzed using Hoechst staining and Annexin V-FITC/PI and JC-1 staining.The anti-tumor action of recombinant adenoviruses on HepG-2 cells was analyzed by constructing a tumor-bearing model in nude mice.Results Ad-VT effectively inhibited the proliferation of HepG-2 cells with a certain time-effect and dose-effect relationship.At a dose of 100 MOI,the rate of inhibition peaked at 64.22%at 72 h,which was significantly higher than that for the control viruses Ad-T(50.52%)and Ad-vp3(43.17%)(P<0.05).Ad-VT causes the death of HepG-2 cells via an endogenous apoptotic pathway.Quantitative analysis of the apoptosis of HepG-2 cells induced by Ad-VT indicated that the rate of apoptosis of HepG-2 cells infected with Ad-VT,Ad-T,and Ad-vp3 was significantly higher than that in the Ad-mock group and the control group after 12 h,and the rate of apoptosis increased gradually over time.The rate of apoptosis induced by Ad-VT peaked at 57.24%at 48 h.Forty-eight h after JC-1 staining,Ad-VT induced the apoptosis of more cells,and the ratio of red fluorescence to green fluorescence decreased significantly(P<0.05).There was no marked changes in the mitochondrial membrane potential in the Ad-mock group and the control group.In in vivo anti-tumor experiments,Ad-VT effectively inhibited tumor growth;it prolonged the survival time and it improved the survival rate of mice.Among all treatment groups,the average rate of tumor inhibition was highest in the Ad-VT treatment group,reaching 76.38%.The average survival time of mice treated with Ad-VT was 41.7 d,which was longer than that of nude mice treated with Ad-T(39.2 d)or Ad-vp3(
作者
李雅茹
杨霞
朱羿龙
李文杰
刘子睿
尚超
宋高杰
李善智
修志儒
葛晨晨
李霄
李一权
金宁一
蔡锦顺
LI Ya-ru;YANG Xia;ZHU Yi-long;LI Wen-jie;LIU Zi-rui;SHANG Chao;SONG Gao-jie;LI Shan-zhi;XIU Zhi-ru;GE Chen-chen;LI Xiao;LI Yi-quan;JIN Ning-yi;CAI Jin-shun(College of Agricuiture,Yanbian University,Yanbian,Jilin 133002,China;Changchun Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Changchun 130122,China;Academicians Workstation of Jilin Province of Changchun University of Chinese Medicine,Changchun 130117,China)
出处
《中国病原生物学杂志》
CSCD
北大核心
2021年第12期1398-1403,共6页
Journal of Pathogen Biology
基金
吉林省青年科技人才托举工程项目(No.QT202111)。