近日，北京生命科学研究所（NIBS）高绍荣实验室与北京蛋白质中心等合作在Proteomics杂志在线发表题为“Embryonic stem cells derived from somatic cloned and fertilized blastocysts are post-transcriptionally indistinguishable: A MicroRNA and protein profile comparison”的文章。该文章报道了使用miRNA芯片和蛋白质组技术分析了体细胞克隆胚胎干细胞和正常受精胚胎干细胞在miRNA水平和蛋白质表达水平，证明体细胞克隆胚胎干细胞与正常受精胚胎干细胞在转录后水平高度类似。
体细胞核移植可以利用特异个体的分化细胞构建克隆胚胎从而可能得到病人特异的核移植胚胎干细胞系，已有的报道已经在小鼠和非人灵长类中基于这种技术成功实施了治疗性克隆。但是由于在克隆动物中发现了大量严重的异常表型，导致了人们对于治疗性克隆在安全性上的担心。虽然在转录水平和发育潜力上，克隆与正常受精的胚胎干细胞系没有明显差异，但是在转录后水平上的系统比较还是空白。为了鉴定克隆和正常受精的胚胎干细胞在转录后水平上是否存在差异，我们通过miRNA芯片、2-D DIGE和生物信息学方法，比较了5个克隆和相对应的正常胚胎干细胞系的miRNA和蛋白质的表达丰度。进一步用stem-loop RT-PCR检测特异miRNAs的表达水平，运用质谱分析技术进一步鉴定差异表达蛋白。我们的结果表明，在miRNA和蛋白质表达丰度上，克隆和正常受精的胚胎干细胞是高度类似的，这些结果与它们类似的发育潜力和转录水平是一致的，进一步证明克隆胚胎干细胞和正常受精来源的胚胎干细胞具有高度类似的治疗潜力。
北京生命科学研究所丁俊军，北京蛋白质中心的郭元彪博士为文章共同第一作者。高绍荣博士为论文的通讯作者。此项研究为科技部863项目和北京市科委资助。(生物谷Bioon.com)
生物谷推荐原始出处：
PROTEOMICS 22 Apr 2009 DOI:10.1002/pmic.200800824
Embryonic stem cells derived from somatic cloned and fertilized blastocysts are post-transcriptionally indistinguishable: A MicroRNA and protein profile comparison
Junjun Ding 1 2, Yuanbiao Guo 3, Sheng Liu 1, Yujuan Yan 3, Gang Chang 1, Zhaohui Kou 1, Yu Zhang 1, Ying Jiang 3, Fuchu He 3, Shaorong Gao 1 *, Jianli Sang 2
1National Institute of Biological Sciences (NIBS), Beijing, P. R. China
2College of Life Science, Beijing Normal University, Beijing, P. R. China
3Beijing Proteome Research Center, Beijing, P. R. China
*Correspondence to Shaorong Gao, #7 Science Park Road, Zhongguancun Life Science Park, Beijing 102206, P. R. China
Therapeutic cloning, whereby somatic cell nuclear transfer is used to generate customized embryonic stem cells (NT-ES) from differentiated somatic cells of specific individuals, has been successfully performed in mice and non-human primates. Safety concerns have prevented this technology from being potentially applied to humans, as severely abnormal phenotypes have been observed in cloned animals. Although it has been demonstrated that the transcriptional profiles and developmental potentials of ES cells derived from cloned blastocysts are identical to those of ES cells derived from fertilized blastocysts (F-ES), a systematic analysis of the post-transcriptional profiles of NT-ES cell lines has not yet been performed. To investigate whether NT-ES cells are comparable to F-ES cells post-transcriptionally, we compared the microRNA and protein profiles of five NT- and matching F-ES cell lines by microRNA microarray, 2-D DIGE and bioinformatic analyses. Stem-loop real-time PCR and MS/MS assays were further performed to verify the expression of specific microRNAs and characterize differentially expressed proteins. Our results demonstrate that the ES cell lines derived from cloned and fertilized mouse blastocysts have highly similar microRNA and protein expression profiles, consistent with their similar developmental potentials and transcriptional profiles.
