目的 :研究核型VASH2在人胚胎组织增殖中的功能。方法 :免疫荧光分析VASH2蛋白在人胚肾细胞293T的表达水平和细胞中的表达部位。利用免疫组化法分析VASH2在多种胚胎组织中的的表达水平。MTT法检测核型VASH2对293T细胞活性的影响,流式细胞仪检测细胞周期。结果 :通过免疫荧光试验发现VASH2蛋白分胞浆型及胞核型2型。胞核型VASH2除了在已经发育完善的心脏组织中阴性表达或弱表达,在其他胚胎组织中显著高表达。核型VASH2的表达与胚胎细胞核分裂指数明显正相关(r=0.826,P〈0.01)。转染核型VASH2可促进293T细胞的生长;转染核型VASH2细胞的G2+S期较空白组及干扰组增加,差异有显著性(P〈0.05)。结论 :VASH2分为胞浆型及核型,核型VASH2具有促进胚胎细胞增殖的作用。
Objective:Vasohibin (VASH),one of new gene families,is believed to be involved in cancer-promoting. However,the exact function is still unknown. To analyze the expression level of nuclear VASH2 protein in human fetal tissues from different gestational ages. Metbods:Immunofluorescent method(IF) was used to detect the expression of VASH2 protein in 293T and intracellular localization. Nuclear VASH2 polyclonal antibody was used to analyze the expression level of nuclear VASH2 protein in human fetal tissues from different gestational ages,and the change of embryonic cell mitotic counts and the expression of nuclear VASH2 was assessed by using immunohistochemistry (IHC). The cell growth was detected by methyl thiazolyl tetrazolium (MTF). Flow Cytometry (FCM) was used to determine the cell cycles. Results:We found that the protein expression of VASH2 (with 355 amino acid residues) was located in the cytoplasm while VASH2 (with 311 amino acid residues) was located in the nucleus. Nuclear VASH2 was generally lower in fully-developed embryos and markedly expression during undergoing several maturational processes. There is a significant positive correlation between nuclear VASH2 and mitotic counts (r = 0.826,P 〈 0.01). Nuclear VASH2 promoted proliferation of 293T cells. FCM analysis showed increasing proportion of S and G2 / M phase in nuclear VASH2 transfection. Conclusion:This is the first study to report differences in the intraeellular localization of VASH2 protein. Nuclear VASH2 protein found in a variety of tissues is not a tissue-specific protein, and mac play imoortant roles in histogenesis and oranoenesis.