Huang Chuan-En, Hu Fang-Wei, Yu Chuan-Hang, Tsai Lo-Lin, Lee Tzu-Hsin, Chou Ming-Yung, Yu Cheng-Chia
School of Dentistry, Chung Shan Medical University, No. 110, Sec. 1, Jianguo N. Rd., Taichung 40201, Taiwan.
Int J Mol Sci. 2014 Oct 15;15(10):18623-39. doi: 10.3390/ijms151018623.
Human dental pulp stem cells (DPSCs), unique mesenchymal stem cells (MSCs) type, exhibit the characteristics of self-renewal and multi-lineage differentiation capacity. Oct4 and Nanog are pluripotent genes. The aim of this study was to determine the physiological functions of Oct4 and Nanog expression in DPSCs. Herein, we determined the critical role of an Oct4/Nanog axis modulating MSCs properties of DPSCs by lentiviral-mediated co-overexpression or co-knockdown of Oct4/Nanog in DPSCs. MSCs properties including osteogenic/chondrogenic/adipogenic induction differentiation was assayed for expression of osteogenic/chondrogenic/adipogenic markers by quantitative real-time RT-PCR analysis. Initially, we observed that the expression profile of Oct4 and Nanog in dental pulp cells, which exerted properties of MSCs, was significantly up-regulated compared to that of STRO-1-CD146- dental pulp cells. Down-regulation of Oct4 and Nanog co-expression significantly reduced the cell proliferation, osteogenic differentiation capability, STRO-1, CD146, and Alkaline phosphatase (ALP) activity of DPSCs. In contrast, co-overexpression of Oct4 and Nanog enhanced the expression level of STRO-1 and CD146, proliferation rate and osteogenic/chondrogenic/adipogenic induction differentiation capability, and expression of osteogenic/chondrogenic/adipogenic induction differentiation markers. Our results suggest that Oct4-Nanog signaling is a regulatory switch to maintain properties in DPSCs.
人牙髓干细胞(DPSCs)是一种独特的间充质干细胞(MSCs)类型,具有自我更新和多向分化能力的特征。Oct4和Nanog是多能基因。本研究的目的是确定Oct4和Nanog在DPSCs中的表达所具有的生理功能。在此,我们通过慢病毒介导的Oct4/Nanog在DPSCs中的共过表达或共敲低,确定了Oct4/Nanog轴调节DPSCs的MSCs特性的关键作用。通过定量实时RT-PCR分析,检测包括成骨/软骨生成/脂肪生成诱导分化在内的MSCs特性,以检测成骨/软骨生成/脂肪生成标志物的表达。最初,我们观察到,与STRO-1-CD146-牙髓细胞相比,具有MSCs特性的牙髓细胞中Oct4和Nanog的表达谱显著上调。Oct4和Nanog共表达的下调显著降低了DPSCs的细胞增殖、成骨分化能力、STRO-1、CD146和碱性磷酸酶(ALP)活性。相反,Oct4和Nanog的共过表达提高了STRO-1和CD146的表达水平、增殖率以及成骨/软骨生成/脂肪生成诱导分化能力,以及成骨/软骨生成/脂肪生成诱导分化标志物的表达。我们的结果表明,Oct4-Nanog信号是维持DPSCs特性的调节开关。