摘要
为探讨高胆固醇致动脉粥样硬化的分子机理 ,利用抑制消减杂交技术克隆胆固醇诱导脐静脉内皮细胞产生的差异表达基因 ,并对其表达进行初步研究。经顺、反两向消减杂交和巢式聚合酶链反应扩增 ,获得了差异表达的cDNA片段 ,克隆化后挑选部分进行测序、同源性比较及应用半定量逆转录—聚合酶链反应分析部分差异基因的表达情况 ,得到 2 3个差异表达的cDNA片段 ,其中 6个新差异表达cDNA片段被GenBank接受。已知基因中的血小板反应蛋白
Aim To investigate cholesterol-induced atherogenesis molecular mechanism, clone and analyse the differentially expressed genes in vascular endothelial cell induced by cholesterol. Methods Suppression subtractive hybridization (SSH) method is used to isolate the differentially expressed cDNA in human umbilical vein endothelial cell induced by cholesterol. After sequencing and homology analysis, ten differentially expressed cDNA fragments are selected and analysed by reverse transcription-polymerase chain reaction(RT-PCR). Results Twenty-three differentially expressed cDNA fragments have been isolated. Seventeen of them are known genes, the other six are unknown genes. The known genes include thrombospondin-1, proteasome subunit (type and so on). The ten elective cDNA fragments (including four known genes and six unknown genes) that are identified by RT-PCR have the same expressed tendency as SSH discovery. Six differentially expressed unknown cDNA fragments have been accepted by GenBank as novel expressed sequence-tags (EST). Conclusion The high level expression of thrombospondin-1 and the low level expression of proteasome subunit (type) in endothelial cell induced by cholesterol may be related to cholesterol-induced atherogenesis.
出处
《中国动脉硬化杂志》
CAS
CSCD
2003年第1期1-4,共4页
Chinese Journal of Arteriosclerosis
基金
国家重点项目研究发展规划 (G2 0 0 0 0 5 70 0 7)资助
关键词
分子生物学
血管内皮细胞
基因差异表达
抑制消减杂交
聚合酶链反应
胆固醇
血小板
反应蛋白
Suppression Subtractive Hybridization
Polymerase Chain Reaction
Cholesterol
Thrombospondin-1
Proteasome Subunit
Endothelial Cell, Human Umbilical Vein
Gene Expression
