您的位置:首页  > 论文页面

Ddx5在胚胎干细胞及重编程中的功能研究

发表时间:2018-02-11  浏览量:454  下载量:98
全部作者: 郭菲菲,康岚
作者单位: 大连医科大学肿瘤干细胞研究院
摘 要: 目的:研究Ddx5 在胚胎干细胞(embryonic stem cell,ESC)、诱导多能干细胞(induced pluripotent stem cell,iPSC)中的作用以及在干细胞、重编程中对下游基因的选择性剪接作用。方法:PCR 检测Ddx5 在ESC及其分化过程中的表达水平和在重编程过程中的基因表达变化;通过在ESC 中过表达和敲低Ddx5,检测其对ESC 多能性的影响;在体细胞重编程中检测Ddx5 的过表达和敲低引起的效率变化,利用流式细胞术和碱性磷酸酶(alkaline phosphatase,AP)染色检测重编程效率;最后利用PCR 检测,分析Ddx5 对下游基因的选择性剪接方式的影响。结果:Ddx5 在ESC 中富集并随分化降低。在ESC 中过表达Ddx5 后,Nanog、Eras 等多能性基因表达上调,敲低则相反。Ddx5 在重编程过程中表达上调,其过表达促进重编程,敲低则明显抑制。Ddx5过表达引起Cd44、Hras 等基因的选择性剪接变化。结论:Ddx5 在ESC 和重编程过程中都发挥着作用,Ddx5过表达引起选择性剪接变化。
关 键 词: 分子生物学;胚胎干细胞;重编程;Ddx5;Ddx17;选择性剪接
Title: Function of Ddx5 in embryonic stem cells and reprogramming
Author: GUO Feifei, KANG Lan
Organization: Institute of Cancer Stem Cell, Dalian Medical University
Abstract: Objective: To investigate the roles of Ddx5 in embryonic stem cell (ESC) cell and induced pluripotent stem cell (iPSC) cell and study the functions of them in alternative splicing downstream genes in stem cells and reprogramming process. Methods: We used PCR to examine the mRNA level of Ddx5 in ESC and its differentiation process, and gene expression changes in reprogramming process. By overexpression or knockdown Ddx5 in ESC, we explored its role in ESC pluripotency effects. In addition, we examined the overexpression level of Ddx5 and investigated the efficiency effect of knockdown Ddx5 in somatic cell reprogramming by flow cytometer and alkaline phosphatase (AP) staining. Finally, we tested the alternative splicing changes of downstream genes under Ddx5 overexpression using splicing PCR. Results: Ddx5 is enrich in ESC and reduces during differentiation. The overexpression of Ddx5 in ESC results in the up-regulation of pluripotency genes (e.g. Nanog, Eras), and the knockdown of Ddx5 is opposite. Moreover, the expression of Ddx5 in reprogramming is up-regulation and promotes efficiency of reprogramming while the knockdown of Ddx5 is opposite. In addition, Ddx5 leads to the alternative splicing changes of Cd44, Hras and so on. Conclusion: Ddx5 plays significant roles in ESC and reprogramming. And overexpression of Ddx5 causes the alternative splicing changes.
Key words: molecular biology; embryonic stem cell; reprogramming; Ddx5; Ddx17; alternative splicing
发表期数: 2018年2月第3期
引用格式: 郭菲菲,康岚. Ddx5在胚胎干细胞及重编程中的功能研究[J]. 中国科技论文在线精品论文,2018,11(3):256-264.
 
4 评论数 0
暂无评论
友情链接