血小板源生长因子和血管紧张素Ⅱ对血管平滑肌细胞中细胞周期相关基因表达的影响

目的:观察血小板源生长因子-BB(PDGF-BB)和血管紧张素Ⅱ(AngⅡ)对血管平滑肌细胞(VSMC)中细胞周期相关基因表达影响的异同。方法:第4-6代培养的大鼠胸主动脉平滑肌细胞,AngⅡ10^-6mol/L和PDGF-BB20μg/L刺激静止后的VSMC,24h收集细胞。用TR1zol试剂提取VSMC中的总RNA,然后用DNA酶纯化总RNA,按DNA芯片试剂盒(Atlas^TMHuman CellCycle Arrays,Clontech公司)的步骤用标记探针,并纯化探针,然后杂交、显影,检测细胞周期相关基因的表达。结果:PDGF-BB刺激组cyclinD3mRNA、cyclinG1mRNA、p57mRNA和p16mRNA相对光密度值明显高于AngⅡ组,前者分别为后者的2.67倍、1.59倍、2.47倍、1.78倍,AngⅡ组E2F-3mRNA和DP2mRNA表达量分别低于PDGF-BB组40.34%和54.07%;而p15mRNA、p19mRNA、E2F-1mRNA、E2F-5mRNA以及N-mycmRNA仅在PDGF-BB刺激组表达;p53结合蛋白mRNA相对A值在AngⅡ组高,为PDGF-BB组的1.94倍;PCNAmRNA、c-myc结合蛋白mRNA、p53依赖性细胞生长因子mRNA、cyclinCmRNA、cyclinB1mRNA、E2F-3mRNA表达在PDGF-BB组和AngⅡ组中接近。结论:在PDGF-BB和AngⅡ诱导的VSMC增殖或肥厚过程中,细胞周期相关基因的表达是不完全相同的。

Effects of platelet-derived growth factor and angiotensin

AIM： To investigate the different expressions of cell cycle related genes in hyperplastic and hypertrophic vascular smooth muscle cells caused by platelet-derived growth factor (PDGF-BB) and angiotensinⅡ(AngⅡ). METHODS: Rat aorta media smooth muscle cells were cultured. PDGF-BB and AngⅡ were added into serum-free medium at a concentration of 20 μg/L and 10-6 mol/L , respectively. Vascular smooth muscle cells (VSMCs) were harvested after stimulated for 24 hours. The expression of cell cycle related genes was measured by DNA chips(Atlas cDNA Expression Arrays, Clontech Laboratories, Inc.). RESULTS: The expression of cyclin D3 mRNA ，cyclin G1 mRNA，p57 mRNA，p16 mRNA，E2F-3 mRNA and DP2 mRNA were higher in PDGF-BB than those in AngⅡstimulated VSMCs. p15 mRNA，p19 mRNA，E2F-1 mRNA， E2F-5mRNA,and N-myc mRNA were only detected in PDGF-BB stimulated group. But the expression of p53-associated protein mRNA were higher in AngⅡstimulation group. The expression of PCNA mRNA, c-myc binding protein mRNA，p53-dependent cell growth regulater mRNA，cyclinC mRNA, cyclinB1 mRNA, E2F-3mRNA were similar in the two groups. CONCLUSION: The procession of cell cycle relys on the coordination of many regulater molecules expressed in different phases. Our study preliminarily definite the genes that express during PDGF-stimulated VSMC's hyperplasia and Ang II-stimulated VSMC's hypertrophy.