您的位置:首页  > 论文页面

hTERT启动子调控的pEGFP-ShCDglyTK双自杀基因系统对裸鼠肝细胞癌皮下移植瘤杀伤作用研究

发表时间:2017-03-15  浏览量:1067  下载量:178
全部作者: 范文哲,张应强,姚望,卢鸣剑,崔伟,王于,李家平
作者单位: 中山大学附属第一医院肿瘤介入科
摘 要: 目的:探讨端粒酶逆转录酶(human telomerase reverse transcriptase,hTERT)启动子调控的pEGFP-ShCDglyTK自杀基因系统对裸鼠SMMC7721细胞皮下移植瘤的杀伤作用。方法:使用BALB/C裸鼠20只构建SMMC7721细胞皮下移植瘤模型,分别转染pEGFP-C1(对照组)、pEGFP-ShCD(CD组)、pEGFP-ShTK(TK组)及pEGFP-ShCDglyTK(CDglyTK组)质粒,观察转染效率并以逆转录PCR(reverse transcription-PCR,RT-PCR)、Western blotting方法检测目的基因的表达。给予5-氟胞嘧啶(5-fluorocytosine,5-FC)及更昔洛韦(ganciclovir,GCV)处理后,常规测量肿瘤体积、病理分析及免疫组化检查p53蛋白及Bcl-2蛋白变化,比较和分析联合基因与单基因的疗效差异。结果:20只裸鼠肝细胞癌皮下移植瘤顺利构建,所有质粒成功转染。CDglyTK组的肿瘤抑制率为65.7%,较CD组的44.8%、TK组的43.1%高(P<0.05);相对于单CD组及单TK组,病理显示肿瘤坏死在CDglyTK组最明显,突变型p53蛋白和Bcl-2蛋白在免疫组化阳性率分别为39.7%和32.3%,表达最少(P<0.05)。结论:pEGFP-ShCDglyTK系统质粒在裸鼠人肝癌细胞SMMC7721的皮下移植瘤中有特异性杀伤作用。
关 键 词: 医学实验动物学;自杀基因;肝癌细胞;端粒酶
Title: Experimental study of killing effect on the transcriptional targeting gene therapy of double suicide gene driven by hTERT promotor pEGFP-ShCDglyTK in hepatocellular carcinoma subcutaneous transplantation tumor of nude mice model
Author: FAN Wenzhe, ZHANG Yingqiang, YAO Wang, LU Mingjian, CUI Wei, WANG Yu, LI Jiaping
Organization: Department of Interventional Oncology, The First Affiliated Hospital, Sun Yat-sen University
Abstract: Objective: To explore the selective killing effect of transcriptional targeting gene therapy of double suicide gene driven by human telomerase reverse transcriptase (hTERT) promotor pEGFP-ShCDglyTK in SMMC7721 cells subcutaneous transplantation tumor of nude mice model. Methods: SMMC7721 cells subcutaneous transplantation tumor model was constructed in 20 BALB/C nude mice. pEGFP-C1 (control group), pEGFP-ShCD (CD group), pEGFP-ShTK (TK group), pEGFP-ShCDglyTK (CDglyTK group) were transfected to tumor respectively. Reverse transcription-PCR (RT-PCR) and Western blotting were used to inspect expression of suicide genes in tumor. The transfected tumors were treated with the 5-fluorocytosine (5-FC) and ganciclovir (GCV), and the treatment effects were evaluated by the volume measurement, pathological analysis and immunohistochemical examination protein p53 and Bcl-2 changes. Results: All SMMC7721 cells subcutaneous transplantation tumor models were successfully constructed and all plasmids were successfully transfected. Compared to the CD (44.8%) and TK (43.1%) groups, tumor inhibition rate of CDglyTK group was higher (65.7%, P<0.05); HE presented tumor necrosis area of CDglyTK group was more obvious, immunohistochemical mutant p53 protein and Bcl-2 protein expression decreased significantly, positive rate were 39.7% and 32.3% respectively (P<0.05). Conclusion: SMMC7721 subcutaneous transplantation tumor of nude mice model was successfully constructed. pEGFP-ShCDglyTK system plasmid system has highly specific killing effects on hepatoma carcinoma cells in vivo experiments.
Key words: medical laboratorial zoology; suicide gene; hepatoma carcinoma cell; telomerase
发表期数: 2017年3月第5期
引用格式: 范文哲,张应强,姚望,等. hTERT启动子调控的pEGFP-ShCDglyTK双自杀基因系统对裸鼠肝细胞癌皮下移植瘤杀伤作用研究[J]. 中国科技论文在线精品论文,2017,10(5):518-526.
 
0 评论数 0
暂无评论
友情链接