PTPN7和UCK1調控老鼠胚胎幹細胞的自我更新

PTPN7 and UCK1 are essential for mouse embryonic stem cell renewal

吳庭毅(GOH TING YI)

指導教授 ： 曹世明

中山醫學大學/醫學院/微生物免疫研究所/碩士(2013年)

摘要

胚胎幹細胞是具有自我更新與分化三大胚層及各式體細胞的獨特能力 (多能性) 。了解胚胎幹細胞的自我更新以及多能性的機制對於發育學，再生醫學等是非常重要的。呂老師實驗室以4801種不同的short hairpin RNA (shRNA) 進行高通量功能性篩選，以尋找與老鼠胚胎幹細胞自我更新或分化有關的kinases 或phosphatases。我們發現了132個並未曾在胚胎幹細胞中被研究過的kinases與phosphatase可以調控老鼠胚胎幹細胞自我更新或分化。其中兩個基因分別是Protein Tyrosine Phosphatase Non-receptor 7 (PTPN7) 與Uridine-Cyditine Kinase 1 (UCK1)。我們以二個不同的shRNA 抑制PTPN7或UCK1的表現 ，研究結果發現，抑制PTPN7以及UCK1會導致胚胎幹細胞的分化、細胞數量大量下降、並抑制與幹細胞自我更新有關的標誌蛋白或樞紐蛋白如Oct4、 Sox2、 Nanog、pSTAT3、STAT3、pAKT、AKT、pERK、和 ERK的表現。這些發現顯示了PTPN7和UCK1在調控胚胎幹細胞中扮演著不可或缺的角色。

關鍵字

胚胎幹細胞 ； 多潛能性 ； 自我更新 ； PTPN7 ； UCK1 ； Oct4 ； Sox2

並列摘要

Embryonic stem cells have a unique property of self-renewal and ability to develop into all three germ layers and different types of somatic cells. An understanding of the molecular mechanism how embryonic stem cells regulate self-renewal and pluripotency are important for developmental biology and regenerative medicine. By a systematically kinase/phosphatase short hairpin RNA (shRNA) functional screen, Lu’s lab identified 132 novel genes regulate self-renewal and differentiation of mouse embryonic stem cells. In these 132 novel genes, the knockdown of PTPN7 (Protein Tyrosine Phosphatase non-receptor type 7) and UCK1 (Uridine-Cytidine Kinase 1) by two different shRNAs induced morphological changes, decreased the cell number, downregulated the expression of the stem cell marker and key pluripotency genes, such as Oct4, Sox2, Nanog, pSTAT3, STAT3, pAKT, AKT, pERK, and ERK. The overexpression of PTPN7 can increase the cell number mouse embryonic stem cell in certain culture conditions. These results revealed the importance of PTPN7 and UCK1 in embryonic stem cell renewal.

並列關鍵字

Embryonic stem cells ； self-renewal ； pluripotency ； PTPN7 ； UCK1 ； Oct4 ； Sox2

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