IGF2基因印记丢失对胚胎干细胞向胰岛样细胞诱导分化的影响

发布时间：2018-03-21 05:53

本文选题：胚胎干细胞　切入点：基因组印迹　出处：《中国组织工程研究》2017年09期 　论文类型：期刊论文

【摘要】：背景:IGF2是一种重要的胚胎有丝分裂生长促进因子,在维持细胞正常生长过程中起关键作用,其基因表达以基因组印迹方式受表观遗传调控。当IGF2发生印迹丢失时,与个体的非正常发育以及肿瘤发生存在一些联系。目的:探讨IGF2基因发生印记丢失对小鼠胚胎干细胞定向诱导分化为胰岛样细胞的影响。方法:体外诱导2种小鼠胚胎干细胞(SF1-G和SF1-1)向胰岛样细胞分化;Real-Time PCR和细胞免疫荧光检测Insulin基因的表达;聚合酶链式反应-限制性内切酶片段长度多态性(PCR-RFLP)检测印迹基因IGF2在诱导分化前后细胞中的亲本表达;体外胰岛素释放实验检测诱导终末细胞的胰岛素释放水平。结果与结论:(1)IGF2基因印记正常(SF1-G)和印记丢失(SF1-1)的2种小鼠胚胎干细胞在诱导分化前后印迹状态没有发生改变;(2)在诱导的终末细胞中,尽管2种细胞的胰岛素释放水平差异无显著性意义,但是Insulin基因在SF1-1组中的表达水平要高于其在SF1-G组的表达水平;(3)IGF2基因印记丢失可能与诱导分化来源的终末细胞中Insulin基因的表达上调相关。
[Abstract]:Background: IGF2 is an important growth promoter of embryonic mitosis, which plays a key role in maintaining normal cell growth. Its gene expression is regulated by epigenetic imprinting. When IGF2 is lost, its gene expression is regulated by epigenetic imprinting. Objective: to investigate the effect of loss of IGF2 gene imprinting on the directional differentiation of mouse embryonic stem cells into islet like cells. Methods: two kinds of small cells were induced in vitro. Murine embryonic stem cells (SF1-G and SF1-1) differentiated into islet like cells with Real-Time PCR and immunofluorescence assay to detect the expression of Insulin gene. Polymerase chain reaction-restriction endonuclease fragment length polymorphism (PCR-RFLP) was used to detect the parental expression of imprinted gene IGF2 in cells before and after differentiation. In vitro insulin release assay was used to detect the level of insulin release in the induced terminal cells. Results and conclusion the two mouse embryonic stem cells with normal and lost imprinting of IGF2 gene were not imprinted before and after differentiation. In induced terminal cells, Although there was no significant difference in insulin release between the two cells, However, the expression level of Insulin gene in SF1-1 group was higher than that in SF1-G group. The loss of Insulin gene imprinting may be related to the up-regulation of Insulin gene expression in the terminal cells derived from induced differentiation.
【作者单位】：深圳市光明新区人民医院内分泌科;南昌大学第一附属医院内分泌科;南昌大学第一附属医院药学部;
【基金】：江西省教育厅科技项目(GJJ13142)~~
【分类号】：R329.2

【相似文献】