项目名称: HSG诱导血管平滑肌细胞凋亡和抑制增殖的功能序列及肽研究
项目编号: No.30872714
项目类型: 面上项目
立项/批准年度: 2009
项目学科: 生物科学
项目作者: 郭小梅
作者单位: 华中科技大学
项目金额: 30万元
中文摘要: 前期研究证实,增殖抑制基因(HSG)也称为线粒体融合素2基因(Mfn2)具有明显诱导血管平滑肌细胞(VSMC)凋亡和抑制其增殖的作用。我们研究发现去除跨膜区序列后该作用增强 40%。本研究采用 PCR 和基因重组等技术,以其 5'p21RAS特性结构序列为基础,构建 6 个含HSG不同结构序列 cDNA 的重组腺病毒载体,感染培养的大鼠 VSMC,并采用分子生物学技术研究目的片段cDNA对细胞中两条重要的信号通路 Ras-Raf-ERK1/2 和Ras-PI3K-Akt的作用, 来阐明抑制VSMCs增殖和促凋亡的主要分子机制。研究结果证实仅含有 16 个氨基酸序列的p21RAS特性结构是Mfn2 抑制平滑肌细胞增殖和诱导细胞凋亡的最小功能片段,我们根据p21RAS 特性结构人工合成的多肽具有显著抑制VSMCs增殖的作用,该作用主要是通过抑制 Ras-PI3K-Akt 信号通路实现的,这对防治心脑血管增生性疾病具有极其重要的意义,为研究药物涂层球囊和支架提供科学依据。
中文关键词: 线粒体融合素2基因;平滑肌细胞;细胞增殖;凋亡
英文摘要: Our preliminary study confirmed that the mitochondrial fusion 2 gene (Mfn2) could significantly inhibit the proliferation of vascular smooth muscle cells(VSCMs) and prevent the restenosis of vessels after injury.More important, we recently explored that adenovirus-mediated tMfn2 gene(Adv-tMfn2) has more significant effect on inhibition of VSMCs proliferation than Mfn2 gene by 40%.In this study, we use PCR and recombinant DNA technology to build six different structures which are part of the Mfn2 gene sequences.We mainly studied the effect of different cDNAs on the two signaling pathways of Ras-Raf-ERK1/2 and Ras-PI3K-Akt,in order to illuminate the molecular mechnism of anti-proliferative activities.The results suggest that the p21RAS characteristic structure of Mfn2 is the smallest functional fragment,then we synthesized p21RAS peptides which could significantly inhibit the proliferation of VSMCs through the Ras-PI3K-Akt signaling pathway.Our study might provide a novel potential therapeutic target in treating cardiovascular proliferative disorders.
英文关键词: Mfn2;Vascular Smooth Muscle Cells; Proliferation;Apoptosis