遗传 ›› 2011, Vol. 33 ›› Issue (6): 613-619.doi: 10.3724/SP.J.1005.2011.00613

• 研究报告 • 上一篇    下一篇

PTEN抑制胚胎原肠胚形成期EMT的过程

李艳1, 王晓钰1, 王丽京2, 徐涛2, 卢晓晔1, 蔡冬青1, 耿建国2, 杨雪松1   

  1. 1. 暨南大学再生医学教育部重点实验室, 医学院组织与胚胎学系, 广州 510632 2. 广东药学院血管生物学实验室, 广州 510224
  • 收稿日期:2010-10-29 修回日期:2011-01-04 出版日期:2011-06-20 发布日期:2011-06-25
  • 通讯作者: 杨雪松 E-mail:yang_xuesong@126.com
  • 基金资助:

    国家重点基础研究发展计划(编号:2010CB529702), 国家自然科学基金项目(编号:30971493, 编号:31071054)和中央高校基本科研业务费专项资金(编号:21610601)资助

PTEN impedes EMT during chick embryo gastrulation

LI Yan1, WANG Xiao-Yu1, WANG Li-Jing2, XU Tao2, LU Xiao-Ye1, CAI Dong-Qing1, GENG Jian-Guo2, YANG Xue-Song1   

  1. 1. Key Laboratory for Regenerative Medicine Ministry of Education, Division of Histology & Embryology Medical College, Jinan University, Guangzhou 510632, China 2. Vascular Biological Laboratory, Guangdong College of Pharmacy, Guangzhou 510224, China
  • Received:2010-10-29 Revised:2011-01-04 Online:2011-06-20 Published:2011-06-25
  • Contact: YANG Xue-Song E-mail:yang_xuesong@126.com

摘要: PTEN(Phosphatase and tensin homolog)是一种重要的抑癌基因, 具有非常广泛的生物学活性, 例如在细胞的生长发育、迁移、凋亡和信号传导等均发挥重要作用。PTEN基因表达始于在胚胎早期的上胚层, 而后主要出现在神经外胚层和胚胎中胚层结构, 表明PTEN可能参与胚胎早期发育过程的细胞迁移、增殖和分化。文章主要应用在体改变早期胚胎PTEN的表达水平来观察其对上胚层至中胚层细胞转换—EMT (Epithelial-mesenchymal transition)的作用。首先, 原位杂交结果提示, 内源性PTEN表达在原条以及之后的中胚层细胞结构如体节等。在体PTEN转染实验, 体外培养至HH3期的鸡胚胎, 转染Wt PTEN-GFP或移植Wt PTEN-GFP原条组织至未转染的同时期的宿主胚胎相同部位后, 观察到PTEN转染细胞大都由上胚层迁移至原条并滞留于原条, 不再参与中胚层细胞形成。移植实验也得到相似结果, 发现在Wt PTEN-GFP阳性原条组织移植后很少细胞迁移出原条。另外在原肠胚期PTEN siRNA降调胚胎一侧PTEN基因后, 降调侧中胚层细胞数明显少于正常侧。上述研究结果均提示PTEN基因在胚胎原肠胚期三胚层形成过程中可能具有抑制EMT的作用。

关键词: EMT, PTEN, 基因表达, 细胞迁移, 原肠胚形成

Abstract: PTEN has been considered as one of the important anti-oncogenes, which possesses very wide biological activities. Endogenous PTEN genes begin to express in epiblast during chick embryo gastrulation, and then the expression extends to neural plate and mesoderm. This suggests that PTEN might be involved in cell migration, proliferation, and differentiation during early embryo development. In this study, we employed in vivo approach to explore if endogenous PTEN participates in EMT (Epithelial-mesenchymal transition) in early chick embryo. PTEN was initially detected to highly express in primitive streak during chick gastrulation, in which EMT occurs, and subsequently mesoderm structure such as somites etc. Then, overexpression of both Wt PTEN-GFP and Wt PTEN-GFP positive transplantation of primitive streak resulted in cell accumulation in primitive streak in the development hereafter, indicating that EMT was blocked in both of our assays, either whole embryo transfection of Wt PTEN-GFP or transplanta-tion of Wt PTEN-GFP primary streak tissue. Finally down-regulation of PTEN gene in one side using PTEN siRNA led to reduce the more number of mesoderm cells in PTNE siRNA side than normal side, which suggests that PTEN gene is probably involved in regulating EMT process in development of early embryonic gastrulation.

Key words: EMT, PTEN, gene expression, cell migration, gastrulation