Ku Bo Mi, Yune Young Phil, Lee Eun Shin, Hah Young-Sool, Park Jae Yong, Jeong Joo Yeon, Lee Dong Hoon, Cho Gyeong Jae, Choi Wan Sung, Kang Sang Soo
Department of Anatomy & Neurobiology, Institute of Health Sciences, School of Medicine, Gyeongsang National University, Jinju 660-290, Republic of Korea.
Department of Rehabilitation Medicine, School of Medicine, Gyeongsang National University, Jinju 660-290, Republic of Korea.
Dev Reprod. 2013 Dec;17(4):299-309. doi: 10.12717/DR.2013.17.4.299.
Transforming growth factor (TGF) family is well known to induce the chondevrepogenic differentiation of mesenchymal stem cells (MSC). However, the precise signal transduction pathways and underlying factors are not well known. Thus the present study aims to evaluate the possible role of C2 domain in the chondevrepogenic differentiation of human mesenchymal stem cells. To this end, 145 C2 domains in the adenovirus were individually transfected to hMSC, and morphological changes were examined. Among 145 C2 domains, C2 domain of protein kinase C eta (PKCη) was selected as a possible chondevrepogenic differentiation factor for hMSC. To confirm this possibility, we treated TGFβ3, a well known chondevrepogenic differentiation factor of hMSC, and examined the increased-expression of glycosaminoglycan (GAG), collagen type II (COL II) as well as PKCη using PT-PCR, immunocytochemistry and Western blot analysis. To further evaluation of C2 domain of PKCη, we examined morphological changes, expressions of GAG and COL II after transfection of PKCη -C2 domain in hMSC. Overexpression of PKCη-C2 domain induced morphological change and increased GAG and COL II expressions. The present results demonstrate that PKCη involves in the TGF-β3-induced chondevrepogenic differentiation of hMSC, and C2 domain of PKCη has important role in this process.
众所周知，转化生长因子（TGF）家族可诱导间充质干细胞（MSC）向软骨细胞分化。然而，其确切的信号转导途径和潜在因素尚不清楚。因此，本研究旨在评估C2结构域在人骨髓间充质干细胞向软骨细胞分化过程中的可能作用。为此，将腺病毒中的145个C2结构域分别转染到人骨髓间充质干细胞中，并观察其形态变化。在145个C2结构域中，蛋白激酶C η（PKCη）的C2结构域被选为可能的人骨髓间充质干细胞软骨细胞分化因子。为证实这一可能性，我们用已知的人骨髓间充质干细胞软骨细胞分化因子TGFβ3处理细胞，并通过PT-PCR、免疫细胞化学和蛋白质印迹分析检测糖胺聚糖（GAG）、II型胶原（COL II）以及PKCη的表达增加情况。为进一步评估PKCη的C2结构域，我们检测了人骨髓间充质干细胞转染PKCη -C2结构域后的形态变化、GAG和COL II的表达。PKCη-C2结构域的过表达诱导了形态变化，并增加了GAG和COL II的表达。目前的结果表明，PKCη参与了TGF-β3诱导的人骨髓间充质干细胞向软骨细胞的分化，并且PKCη的C2结构域在此过程中起重要作用。
