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.
出处
《中华眼科杂志》
CAS
CSCD
北大核心
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)
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
Transcription factors
Activin receptors, type Ⅰ
Angiogenesis inducing agents
Vascular endothelial growth factor receptor-2
