组蛋白去乙酰化抑制剂largazole在抗hUCMSCs衰老中的作用  

The role of histone deacetylase inhibitor largazole in anti-aging of hUCMSCs

作　　者：韩保卫[1] 王云帅[1,2] 闫红杰[2] 

机构地区：[1]郑州大学附属洛阳中心医院普外科,河南洛阳471000 [2]暨南大学附属第二临床学院普外科,广东深圳518001

出　　处：《基础医学与临床》2015年第3期387-392,共6页Basic and Clinical Medicine

摘　　要：目的采用组蛋白去乙酰化抑制剂largazole作用于人脐带间充质干细胞(h UCMSCs),探讨largazole在防止h UCMSCs衰老作用和机制。方法分离、纯化、鉴定人脐带间充质干细胞(h UCMSCs),筛选组蛋白去乙酰化抑制剂largazole扩增h UCMSCs合适的作用浓度。利用合适浓度进行h UCMSCs传代,通过real-time PCR法检测largazole组P4、P 8、P12和P16代h UCMSCs干性基因表达水平;通过CHIP检测组蛋白乙酰化抑制剂largazole对h UCMSCs增殖、分化相关基因(OCT4、TERT、OPN)组蛋白H3K9、H3K14的乙酰化程度的影响。结果 h UCMSCs体外扩增随着传代数的增加,增殖能力逐渐下降,干性基因的表达水平也随之降低,并表现出来向成骨方向自主分化。低剂量的largazole可提高h UCMSCs增殖能力,延缓h UCMSCs的衰老。Largazole主要通过修饰OCT4、TERT、OPN基因启动子区域的组蛋白H3K9/K14ac,使h UCMSCs细胞增殖(OCT4)和端粒酶基因(TERT)mRNA表达增强,成骨分化基因(OPN)的表达减弱。结论表明组蛋白去乙酰化抑制剂largazole可以通过修饰h UCMSCs组蛋白H3的乙酰化,在调控MSCs的生物学特性方面起到重要的作用。Objective This research adopt the histone acetylation inhibitor largazole to treat on human umbilical cord mesenchymal stem cells（ h UCMSCs）,and to discuss the role and molecular mechanism of anti-aging.Methods h UCMSCs were isolated from human Umbilical cords after trypsin digestion,and determined by Flow cytometry analysis. they were cultured with different concentrations of largazole to find the fine cell proliferative concentrations. Passage 4,8,12,and 16 cells were subjected to real-time PCR for the change of stem cell gene expression on largazole group. We investigated the effect of largazole,against the histone H3 lysine 9 or 14（ H3K9 /K14） acetylation of the gene（ TERT,OCT4 and OPN） by CHIP assays. Results h UCMSCs can undergo spontaneous differentiation and aging during expansion. Low concentrate largazole can promote h UCMSCs proliferation and suppresse its spontaneous osteogenic differentiation. Largazole modulates histone H3 acetylation in the TERT,OCT4 and OPN genes. Conclusions Histone H3 acetylation,which can be modulated by largazole,plays a key role in regulating h UCMSCs biological characteristics.

关 键 词：干细胞 增殖 生物学特性 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]