吴峰 冯金忠 李松华 俞峰 邱一华 张帅 周巍 骆合德 王国坤.热休克蛋白90 对血小板源衍生因子诱导的大鼠主动脉平滑肌细胞增殖的影响[J].,2014,14(31):5062-6064 |
热休克蛋白90 对血小板源衍生因子诱导的大鼠主动脉平滑肌细胞增殖的影响 |
Effect of Heat Shock Protein 90 on Proliferation of Rat Aortic SmoothMuscle Cells Induced by Platelet Derived Growth Factor |
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DOI: |
中文关键词: 热休克蛋白90 平滑肌细胞 增殖 细胞周期 |
英文关键词: Heat shock protein 90 Smooth muscle cells Proliferation Cell cycle |
基金项目:上海市自然科学基金项目(13ZR1409000) |
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中文摘要: |
目的:研究热休克蛋白90(HSP90)对血小板衍生因子(Platelet derived growth factor,PDGF)诱导的大鼠主动脉平滑肌细胞增
殖的影响。方法:采用胶原酶消化法原代培养大鼠胸主动脉平滑肌细胞,应用脂质体细胞转染siRNA的方法抑制HSP90 的表达,
定量PCR和western blot的方法检测抑制效率。利用PDGF-bb 诱导刺激平滑肌细胞增殖,CCK8 法检测细胞增殖能力的变化,流
式细胞术检测细胞生长周期的改变。结果:平滑肌细胞中转染HSP90 的siRNA后,HSP90 的mRNA和蛋白水平明显降低,分别
为对照组的65.3 %和57.6 %(P<0.05);PDGF-bb 刺激明显促进平滑肌细胞生长,而降低HSP90 水平显著影响PDGF-bb 诱导的细
胞增殖(P<0.05);流式细胞术检测发现降低HSP90 水平引起平滑肌细胞生长停滞,分布在细胞周期G1 期的细胞比例明显增多
(P<0.05)。结论:HSP90 通过调控平滑肌细胞的生长周期参与调节细胞增殖过程。 |
英文摘要: |
Objective:To investigate the effect of heat shock protein 90 (HSP90) on proliferation of rat aortic smooth muscle cells
induced by PDGF (Platelet derived growth factor).Methods:The primary aortic smooth muscle cells were cultured from thoracic aortas
by collagenase digestion methods. The expression of HSP90 was intervened by liposomes with siRNA, and determined by real-time PCR
and western blot. Proliferation of smooth muscle cells were stimulated PDGF-bb, and were detected by CCK8 assay. The cell cycle of
smooth muscle cells were determined by flow cytometry.Results:Smooth muscle cells transfected with siRNA 24 h later, mRNA and
protein levels of HSP90 significantly decreased by 65.3% and 57.6% respectively, compared with control groups. The proliferation of
fibroblasts were markedly promoted by PDGF-bb, but HSP90 siRNA suppressed the cell proliferation. Flow cytometry assay showed that
smooth muscle cells transfected with HSP90 siRNA were found arrest in the G1 phase of the cell cycle.Conclusion:HSP90 is involved
in the proliferation of smooth muscle cells by regulating cell cycle. |
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