兰州大学机构库 >基础医学院
Survivin-siRNA真核表达载体的构建及其对胃癌SGC-7901细胞增殖和凋亡的影响
Alternative TitleConstruction of Eukaryotic Expression Vector for Survivin-siRNA and Its Effect on Proliferation and Apoptosis of Gastric Cancer SGC-7901 Cells
陈卫; 王玉平; 庞春艳; 王毅君
2009-10-20
Source Publication中国生物制品学杂志   Impact Factor & Quartile Of Published Year  The Latest Impact Factor & Quartile
ISSN1004-5503
Volume22Issue:10Pages:975-978+982
Abstract目的构建Survivin-siRNA真核表达载体,并探讨其对胃癌SGC-7901细胞增殖和凋亡的影响。方法化学合成4条能转录出siRNA的模板DNA,各75个碱基,退火形成2条双链DNA,双酶切后插入pSUPER.basic载体。将阳性重组质粒转染SGC-7901细胞,进行细胞计数,并用MTT法检测细胞的增殖活性;半定量RT-PCR检测细胞Survivin基因mRNA的转录水平;流式细胞术检测细胞周期的变化。结果PCR和酶切鉴定表明Survivin-siRNA真核表达载体构建正确,其能下调SGC-7901细胞Survivin基因mRNA的转录水平,抑制SGC-7901细胞生长和增殖,并促进细胞...
Other AbstractObjective To construct a eukaryotic expression vector for survivin-siRNA and investigate its effect on proliferation and apoptosis of gastric cancer SGX-7901 cells. Methods Four single-stranded template DNAs encoding siRNA against survivin, each consisting of 75 bp, were synthesized chemically, based on which two double-stranded DNAs were formed by annealing, then identified by restriction analysis and inserted into vector pSUPER.basic. SGC-7901 cells were transfected with positive recombinants, then counted and determined for proliferation activity by MTT method, and for cell cycle by flow cytometry. The transcription level of Survivin mRNA was determined by semi-quantitative RT-PCR. Results Both PCR and restriction analysis proved that the eukaryotic expression vector for Survivin-siRNA was constructed correctly. The constructed expression vector down-regulated the transcription level of Survivin mRNA, inhibited the growth and proliferation and promoted the apoptosis of SGC-7901 cells. The number of transfected SGC-7901 cells at G(0)/G(1) and sub-G, stages increased while that at S stage decreased. Conclusion The eukaryotic expression vector for Survivin-siRNA was successfully constructed, which down-regulated the transcription of Survivin mRNA in SGC-7901 cells, and inhibited the proliferation and promoted the apoptosis of the cells. It provided a certain experimental basis for gene therapy of gastric cancer by RNAi technique.
KeywordSurvivin 小干扰RNA RNA干扰 真核表达载体 胃癌 增殖 凋亡
Subject AreaScience & Technology - Other Topics ; Oncology ; Genetics & Heredity ; Gastroenterology & Hepatology
Indexed ByBIOSIS ; CSCD
Language中文
Funding Project甘肃省自然科学基金计划
Project Number甘肃省自然科学基金项目资助(3ZS051-A25-079)
CSCD IDCSCD:3699438
Funding OrganizationGSSTD
BIOSIS IDBIOSIS:PREV201000002036
IRIDCNKI:0037614
Department兰州大学基础医学院生物化学与分子生物学研究所
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Document Type期刊论文
Identifierhttps://ir.lzu.edu.cn/handle/262010/163109
Collection基础医学院
Recommended Citation
GB/T 7714
陈卫,王玉平,庞春艳,等. Survivin-siRNA真核表达载体的构建及其对胃癌SGC-7901细胞增殖和凋亡的影响[J]. 中国生物制品学杂志,2009,22(10):975-978+982.
APA 陈卫,王玉平,庞春艳,&王毅君.(2009).Survivin-siRNA真核表达载体的构建及其对胃癌SGC-7901细胞增殖和凋亡的影响.中国生物制品学杂志,22(10),975-978+982.
MLA 陈卫,et al."Survivin-siRNA真核表达载体的构建及其对胃癌SGC-7901细胞增殖和凋亡的影响".中国生物制品学杂志 22.10(2009):975-978+982.
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