发状分裂相关增强子-1对血管形成相关因子作用的实验研究  

Gene regulatory mechanism of hairy and enhancer of split related-1 for angiogenesis factors

作　　者：李斌[1] 唐仕波[1] 林少芬[1] 孟晶[1] 

机构地区：[1]中山大学中山眼科中心,广州510060

出　　处：《中华眼科杂志》2006年第2期116-120,共5页Chinese Journal of Ophthalmology

基　　金：国家自然科学基金群体创新资助项目(30321004);国家自然科学基金资助项目(30471849);广东省重点科技基金资助项目(052001144A)

摘　　要：目的研究发状分裂相关增强子1(HESR1)基因在血管形成和维持中的作用。方法体外培养人脐静脉内皮细胞(HUVEC),构建PcDNA3.1+HESR1重组质粒、RNA干扰用pSIREN+HESR1质粒。脂质体将这两个质粒分别转染到HUVEC,并以转染空白质粒体作为对照。在HUVEC内分别高表达HESR1、下调HESR1的表达,采用半定量逆转录聚合酶链反应(RTPCR)和免疫印迹法检测HUVEC内HESR1不同表达状态时血管内皮生长因子第2受体(KDR)、激活素受体样激酶1(ALK1)、血管生成因子1(Ang1)表达的改变。结果在HUVEC内高表达HESR1基因能下调KDR的表达,上调ALK1、Ang1的表达;当抑制HESR1基因,KDR的表达上调,ALK1、Ang1的表达下调。结论HESR1基因可调控HUVEC中KDR、ALK1、Ang1的表达,在血管的形成和维持中发挥关键作用。Objective To study the roles of hairy and enhancer of split related-1 （ HESR-1 ） in maintenance of the mature, quiescent vessel and angiogenesis. Methods Human umbilical vein endothelial cells（HUVEC） were cultured. The full-length coding sequence of HESR-1 was cloned into PcDNA3. 1 ＋ using standard protocols. HESR-1 specific siRNA was synthesized and cloned into the RNAi-Ready pSIREN- RetroQ ZsGreen Vector. The constructed PcDNA3.1 ＋ HESR-1 plasmid were transfected into HUVEC for the overexpression of HESR-1, and HESR-1-RNAi plasmid were transfected into HUVEC to silence the HESR-1 gene, the expression of KDR , ALK-1 and Ang-1 in HUVEC were analyzed by RT-PCR and Western blot. Results The expression of KDR was down-regulated and ALK-1 and Ang-1 were up-regulated in HUVEC with the overexpression of HESR-1 ; The expression of KDR was up-regulated and ALK-1 and Ang-1 were down-regulated the HESR-1 in HUVEC by RNAi. Conclusion HESR-1 may play an important role in maintenance of vessel in quiescent and control angiogenesis by the regulation of the expression of KDR ,ALK- 1 and Ang-1.

关 键 词：转录因子 激活素受体 Ⅰ型 血管生成诱导剂 血管内皮生长因子受体2 

分 类 号：R774.1[医药卫生—眼科]