中国组织工程研究 ›› 2017, Vol. 21 ›› Issue (9): 1362-1367.doi: 10.3969/j.issn.2095-4344.2017.09.010

• 胚胎干细胞 embryonic stem cells • 上一篇    下一篇

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

刘  峰1,2,彭宇环3,汤佳珍2,姜  杉2   

  1. 1深圳市光明新区人民医院内分泌科,广东省深圳市  518106;南昌大学第一附属医院,2内分泌科,3药学部,江西省南昌市  330006
  • 出版日期:2017-03-28 发布日期:2017-03-31
  • 作者简介:刘峰,男,1980年生,江西吉安市安福县人,汉族,2010年中南大学毕业,博士,副主任医师,主要从事糖尿病及其并发症的基础与临床工作。
  • 基金资助:

    江西省教育厅科技项目(GJJ13142)

Effect of the imprinting loss of insulin like growth factor 2 gene during the differentiation from mouse embryonic stem cells to islet-like cells in vitro

Liu Feng1, 2, Peng Yu-huan3, Tang Jia-zhen2, Jang Shan2   

  1. 1Department of Endocrinology, People’s Hospital of Guangming New District, Shenzhen 518106, Guangdong Province, China; 2Department of Endocrinology, 3Department of Pharmacy, the Affiliated Hospital of Nanchang University, Nanchang 330006, Jiangxi Province, China
  • Online:2017-03-28 Published:2017-03-31
  • About author:Liu Feng, M.D., Associate chief physician, Department of Endocrinology, People’s Hospital of Guangming New District, Shenzhen 518106, Guangdong Province, China
  • Supported by:

    the Science and Technology Project of Jiangxi Provincial Education Department, No. GJJ13142

摘要:

文章快速阅读:

文题释义:
基因组印记:
是一种表观遗传机制,它可以限制某个基因只能由来自双亲染色体中的一条表达。对于只表达父本、母本关闭的基因称为母本印记基因,如胰岛素样生长因子2;只表达母本、父本关闭的基因称为父本印记基因,如H19。在哺乳动物基因组中,约有1%的基因属于印记基因范畴。目前,在人和小鼠的研究中已鉴定出超过100个印记基因,这些印记基因对于哺乳动物的正常发育至关重要。
IGF2基因与印记丢失:人IGF2基因全长8 837 bp,包括9个外显子和8个内含子。人类有4个启动子P1、P2、P3、P4。在生长发育过程中,4个启动子生物活性各不相同,具有组织特异性,也与发育阶段相关。当基因印迹丢失时,常导致IGF2过表达,并直接引起动物克隆形成与胚胎发育过程的异常以及促进肿瘤细胞的恶性生长。现有许多研究表明IGF2基因印迹丢失已经成为一些肿瘤发生的重要标志。

 

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:①IGF2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;③IGF2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。

 

中国组织工程研究杂志出版内容重点:干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程

ORCID: 0000-0002-8543-6078(刘峰)

关键词: 干细胞, 胚胎干细胞, 基因印记, 诱导分化, 糖尿病, 胰岛素释放, SF1-1, SF1-G, 胰岛相关标记基因

Abstract:

BACKGROUND: Insulin like growth factor 2 (IGF2) is an important embryonic mitosis growth promoting factor, which plays a critical role in the process of maintaining normal cell growth. The gene expression of IGF2 is under epigenetic regulation in the way of genomic imprinting. Imprinting loss of IGF2 is likely to be associated with the abnormal development of the individual and tumorigenesis.
OBJECTIVE: To investigate the effect of imprinting loss of IGF2 gene on the differentiation of mouse embryonic stem cells into islet-like cells.
METHODS: Two kinds of mouse embryonic stem cells (SF1-G and SF1-1) were induced to differentiate into islet-like cells in vitro. The expression of Insulin gene was detected by real-time PCR and cell immunofluorescence. The expression of IGF2 was detected by the polymerase chain reaction-restriction fragment length polymorphism in the cells before and after induced differentiation. The level of insulin released at terminal differentiation stage was tested by insulin release assay in vitro.
RESULTS AND CONCLUSION: (1) There was no change in the imprinting state of the two kinds of mouse embryonic stem cells with normal and imprinted IGF-2 gene before and after differentiation. (2) In the induced cells, the expression level of insulin in the SF1-1 group was higher than that in the SF1-G group, although there was no significant difference in the insulin release between the two kinds of cells. (3)The imprinting loss of IGF-2 gene may be related to the up-regulation of insulin mRNA expression in terminal cells during induced differentiation.

 

 中国组织工程研究杂志出版内容重点:干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程

Key words: Embryonic Stem Cells, Genomic Imprinting, Insulin-Like Growth Factor II, Insulin, Tissue Engineering

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