摘要
目的 克隆、分离血管内皮细胞在致动脉粥样硬化因素作用下差异表达的基因 ,了解动脉粥样硬化发生的分子机制。方法 培养血管内皮细胞系 (ECV30 4)在低密度脂蛋白 (LDL)的诱导下 ,用改进的差异显示 聚合酶链反应 (PCR)技术分析表达水平明显差异的cDNA。对差异显示基因进行序列测定和同源比较。用NorthernBlot证实差异显示基因片段。结果 LDL诱导ECV30 4细胞出现一些上调和下调表达的cDNA。已知的上调表达基因包括 10 0 0 0蛋白、FGFR1原癌基因伴随蛋白、rTSβ蛋白、FK5 0 6结合蛋白、细胞间粘附分子 1;下调表达的基因有fb19、Apobec 1结合蛋白 1、RBP2。不同的上调或下调基因表达水平变化在 70 %～ 2 0 0 %之间不等。结论 用差异显示 PCR方法证实LDL可诱导ECV30 4细胞许多基因表达水平的变化。高水平LDL作用于内皮细胞可引起炎性反应 ,影响动脉粥样硬化的早期发生。
Objective To isolate and clone the differential expressed genes induced by atherogenic factors on vascular endothelium and to understand the molecular mechanism of atherosclerosis. Methods The differential display reverse transcription PCR method (DDRT PCR) was used to analyze the differential expressed cDNA in ECV304 induced by low density lipoprotein (LDL). After sequencing and homology research, the differential expressed cDNA fragments were confirmed by northern blot analysis. Results Up regulated and down regulated cDNAs fragments were isolated in ECV304. It was found that some cDNAs fragments were highly homologous to the known human genes, while others were fragments of novel genes. The known up regulated genes included human 10 kD protein, FGFR1 oncogene partner, intercellular adhesion molecular 1, FK506 binding protein and rTS beta genes. The known down regulated genes included human fb19, Apobec1 binding protein 1 and RBP2. The expression levels of different genes varied by 70%～200%. Conclusion By DDRT PCR method, our results showed that LDL could change the expression of many genes in endothelial cells. The high expression level of ICAM 1 in the endothelial cells stimulated by LDL suggested that ICAM 1 is an important adhesion molecular in atherogenesis. The down regulated expression of Apobec 1 binding protein 1 may affect the cellular synthesis of apolipoprotein. The high expression level of FGFR1 oncogene partner is likely related to the differentiation of ECV304. The function of other differential expressed genes remains to be elucidated.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2000年第10期784-786,共3页
National Medical Journal of China
基金
国家攀登计划基金
