Damiola Francesca, Keime Céline, Gonin-Giraud Sandrine, Dazy Sébastien, Gandrillon Olivier
Equipe 'Signalisations et identités cellulaires', Centre de Génétique Moléculaire et Cellulaire CNRS UMR 5534, France.
Oncogene. 2004 Oct 7;23(46):7628-43. doi: 10.1038/sj.onc.1208061.
The molecular mechanisms regulating the cell fate decision between self-renewal and differentiation/apoptosis in stem and progenitor cells are poorly understood. Here, we report the first comprehensive identification of genes potentially involved in the switch from self-renewal toward differentiation of primary, non-immortalized erythroid avian progenitor cells (T2EC cells). We used the Serial Analysis of Gene Expression (SAGE) technique in order to identify and quantify the genome fraction functionally active in a self-renewing versus a differentiating cell population. We generated two SAGE libraries and sequenced a total of 37,589 tags, thereby obtaining the first transcriptional profile characterization of a chicken cell. Tag identification was performed using a new relational database (Identitag) developed in the laboratory, which allowed a highly satisfactory level of identification. Among 123 differentially expressed genes, 11 were investigated further and for nine of them the differential expression was subsequently confirmed by real-time PCR. The comparison of tag abundance between the two libraries revealed that only a small fraction of transcripts was differentially expressed. The analysis of their functions argue against a prominent role for a master switch in T2EC cells decision-making, but are in favor of a critical role for coordinated small variations in a relatively small number of genes that can lead to essential cellular identity changes.
干细胞和祖细胞中调节自我更新与分化/凋亡之间细胞命运决定的分子机制目前仍知之甚少。在此,我们首次全面鉴定了可能参与原代、非永生化禽类红系祖细胞(T2EC细胞)从自我更新向分化转变过程的基因。我们使用基因表达序列分析(SAGE)技术来鉴定和量化在自我更新细胞群体与分化细胞群体中具有功能活性的基因组部分。我们构建了两个SAGE文库,并对总共37,589个标签进行了测序,从而获得了鸡细胞的首个转录图谱特征。标签鉴定使用了实验室开发的一个新的关系数据库(Identitag),该数据库实现了高度令人满意的鉴定水平。在123个差异表达基因中,有11个被进一步研究,其中9个基因的差异表达随后通过实时PCR得到了证实。两个文库之间标签丰度的比较显示,只有一小部分转录本存在差异表达。对其功能的分析表明,在T2EC细胞的决策过程中,主开关并没有发挥突出作用,而是支持相对少数基因的协调微小变化在导致细胞基本身份改变方面发挥关键作用。