[1]陈建国,卢天龙,韩跃武,等.BAG-1、Bcl-2双靶区反义RNA重组载体诱导SGC-7901细胞凋亡[J].陆军军医大学学报(原第三军医大学学报),2009,31(02):132-135.
　CHEN Jian-guo,LU Tian-long,HAN Yue-wu,et al.Recombinant vector carrying antisense RNA to dual-target BAG-1 and Bcl-2 induces SGC-7901 cell apoptosis[J].J Amry Med Univ (J Third Mil Med Univ),2009,31(02):132-135.
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陆军军医大学学报（原第三军医大学学报）[ISSN:1000-5404/CN:51-1095/R]

卷:
31卷
期数:
2009年第02期
页码:
132-135
栏目:
论著
出版日期:
2009-01-30

文章信息/Info

Title:
Recombinant vector carrying antisense RNA to dual-target BAG-1 and Bcl-2 induces SGC-7901 cell apoptosis
作者:
陈建国; 卢天龙; 韩跃武; 冯惟萍; 任慧子
兰州大学基础医学院医学生物化学与分子生物学研究所; 甘肃省卫生学校
Author(s):
CHEN Jian-guo;  LU Tian-long;  HAN Yue-wu;  FENG Wei-ping;  REN Hui-zi
Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Lanzhou University, Lanzhou 730000; Gansu Health School, Lanzhou 730000, China
关键词:
BAG-1; Bcl-2; 双表达质粒; 反义RNA; 胃癌
Keywords:
BAG-1;  Bcl-2;  co-expression vector;  antisense RNA;  gastric carcinoma
分类号:
R394-33；R73-354；R735.2
文献标志码:
A
摘要:
目的    构建BAG-1、Bcl-2双靶区反义RNA重组载体并初步探讨其对SGC-7901细胞增殖活性的影响。    方法    从SGC-7901细胞总RNA中逆转录扩增包括全部编码序列的BAG-1和Bcl-2 cDNA，利用生物工程技术，反向插入真核细胞双表达载体pVITRO2的多克隆位点中，转染SGC-7901细胞，MTT法检测对细胞增殖的影响，RT-PCR法观察对细胞BAG-1和Bcl-2 mRNA表达水平的变化，流式细胞术检测对细胞周期的影响。    结果    限制性内切酶和测序分析表明，双表达质粒pVITRO2-AsBAG-1-Bcl-2构建成功。MTT法检测显示pVITRO2-AsBAG-1-Bcl-2载体抑制SGC-7901细胞增殖，并呈时间依赖性，其中以72 h转染组抑制作用最显著(P<0.01)；与对照组比较，pVITRO2-AsBAG-1-Bcl-2 组BAG-1和Bcl-2 mRNA表达水平下降(P<0.05)，细胞凋亡比例升高(P<0.01)。    结论    成功构建反义双靶区重组载体pVITRO2-AsBAG-1-Bcl-2，并发现其能抑制SGC-7901细胞增殖并引起细胞凋亡。
Abstract:
Objective    To construct the recombinant co-expression vector carrying antisense RNAs to dual-target BAG-1 and Bcl-2 and investigate its effect on the proliferation of SGC-7901 cells.     Methods    RT-PCR was used to amplify the full length CDS of BAG-1 and Bcl-2 cDNA from total RNA of SGC-7901 cells. The cDNA fragments of BAG-1 and Bcl-2 were inserted into eukaryotic co-expression pVITRO2 vector multiple cloning sites in the antisense orientation respectively by gene recombination technology. Then the recombinant plasmid was transfected into SGC-7901 cell. The proliferation of the transfected SGC-7901 cells was determined by MTT assay. The changes of BAG-1 and Bcl-2 mRNA levels were detected by RT-PCR and the division of cell cycle was detected by FCM.     Results    The restriction endonucleases digestion and sequencing suggested that the eukaryotic co-expression vector pVITRO2-AsBAG-1-Bcl-2 was constructed successfully. MTT assay demonstrated that pVITRO2-AsBAG-1-Bcl-2 could inhibit the proliferation of SGC-7901 cells in a time-dependent manner, with most significant effect in 72 h after transfection (P<0.01). BAG-1 and Bcl-2 mRNA expressions in pVITRO2-AsBAG-1-Bcl-2 transfected cells were significantly decreased compared with those in the control cells and pVITRO2 transfected cells (P<0.05). The apoptotic rate of pVITRO2-AsBAG-1-Bcl-2 transfected cells was significantly higher than that in control cells and pVITRO2 transfected cells (P<0.01).     Conclusion    The co-expression vector pVITRO2-AsBAG-1-Bcl-2 has been successfully constructed and it can inhibit the proliferation of SGC-7901 cells and induce cell apoptosis.

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更新日期/Last Update: 2009-01-08