近日,，北京生命科學(xué)研究所（NIBS）高紹榮實(shí)驗(yàn)室與北京蛋白質(zhì)中心等合作在Proteomics雜志在線發(fā)表題為“Embryonic stem cells derived from somatic cloned and fertilized blastocysts are post-transcriptionally indistinguishable: A MicroRNA and protein profile comparison”的文章,。該文章報(bào)道了使用miRNA芯片和蛋白質(zhì)組技術(shù)分析了體細(xì)胞克隆胚胎干細(xì)胞和正常受精胚胎干細(xì)胞在miRNA水平和蛋白質(zhì)表達(dá)水平，證明體細(xì)胞克隆胚胎干細(xì)胞與正常受精胚胎干細(xì)胞在轉(zhuǎn)錄后水平高度類似,。
體細(xì)胞核移植可以利用特異個(gè)體的分化細(xì)胞構(gòu)建克隆胚胎從而可能得到病人特異的核移植胚胎干細(xì)胞系,，已有的報(bào)道已經(jīng)在小鼠和非人靈長(zhǎng)類中基于這種技術(shù)成功實(shí)施了治療性克隆。但是由于在克隆動(dòng)物中發(fā)現(xiàn)了大量嚴(yán)重的異常表型,，導(dǎo)致了人們對(duì)于治療性克隆在安全性上的擔(dān)心,。雖然在轉(zhuǎn)錄水平和發(fā)育潛力上，克隆與正常受精的胚胎干細(xì)胞系沒有明顯差異,，但是在轉(zhuǎn)錄后水平上的系統(tǒng)比較還是空白,。為了鑒定克隆和正常受精的胚胎干細(xì)胞在轉(zhuǎn)錄后水平上是否存在差異，我們通過(guò)miRNA芯片,、2-D DIGE和生物信息學(xué)方法,，比較了5個(gè)克隆和相對(duì)應(yīng)的正常胚胎干細(xì)胞系的miRNA和蛋白質(zhì)的表達(dá)豐度。進(jìn)一步用stem-loop RT-PCR檢測(cè)特異miRNAs的表達(dá)水平,，運(yùn)用質(zhì)譜分析技術(shù)進(jìn)一步鑒定差異表達(dá)蛋白,。我們的結(jié)果表明，在miRNA和蛋白質(zhì)表達(dá)豐度上,，克隆和正常受精的胚胎干細(xì)胞是高度類似的,，這些結(jié)果與它們類似的發(fā)育潛力和轉(zhuǎn)錄水平是一致的，進(jìn)一步證明克隆胚胎干細(xì)胞和正常受精來(lái)源的胚胎干細(xì)胞具有高度類似的治療潛力,。
北京生命科學(xué)研究所丁俊軍,，北京蛋白質(zhì)中心的郭元彪博士為文章共同第一作者,。高紹榮博士為論文的通訊作者。此項(xiàng)研究為科技部863項(xiàng)目和北京市科委資助,。(生物谷Bioon.com)
生物谷推薦原始出處：
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.
