In vitro induction of differentiation by ginsenosides in F9 teratocarcinoma cells
In vitro induction of differentiation by ginsenosides in F9 teratocarcinoma cells
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DOI：
10.1016/0959-8049(96)00102-5
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发表时间：
1996-07-01
影响因子：
8.4
通讯作者：
Kim, KW
中科院分区：
文献类型：
--
作者：
Lee, YN;Lee, HY;Kim, KW
本研究的目的是确定人参皂苷，人参提取物的能力。Meyer等，以引起F9畸胎瘤支架细胞的分化作为模型系统。F9干细胞培养的存在下的皂苷与二丁酰环AMP（dbcAMP）成为壁内胚层样细胞。此外，分化标记基因的表达，如层粘连蛋白B1和IV型胶原蛋白，增加治疗后的黄芪皂苷。在各种纯化的芦荟苷中，Rh和Rh在引起F9细胞分化方面最有效。由于人参皂苷和糖皮质激素具有相似的化学结构，我们研究了人参皂苷诱导的分化过程中糖皮质激素受体（GR）参与的可能性。根据Southwestern印迹分析，在培养于含有甘草皂苷Rh-1或Rh-2的培养基中的F9细胞中检测到被认为是GR的94 kDa蛋白质。此外，用人参皂苷Rh-1或Rh-2和RU 486（一种对GR具有高亲和力的糖皮质激素拮抗剂）处理的F9干细胞在形态上不分化成内胚层细胞，并且在这些细胞中不诱导层粘连蛋白B1基因的表达。在凝胶迁移率变动分析中，能够特异性结合糖皮质激素反应元件（GRE）的蛋白质因子被检测到在阿糖胞苷处理的F9细胞的核提取物中。此外，GR表达载体和GR-荧光素酶载体共转染的GR过表达增强了GRE启动子的反式激活活性，并进一步增强了双环腺苷酸的存在下，皂苷Rh-1或Rh-2。此外，人参皂苷Rh-1和Rh-2与通过全细胞结合测定评估的GR结合，即使与地塞米松相比特异性结合亲和力较弱。根据这些数据，我们认为，人参皂苷Rh-1和Rh-2引起F9细胞的分化，人参皂苷的作用可能是通过与GR或其类似的核受体结合而发挥的。版权所有（C）1996 Elsevier Science Ltd
The aim of this study was to determine the ability of the ginsenosides, extracts of Panax ginseng C.A. Meyer, to cause differentiation of F9 teratocarcinoma stent cells as a model system. F9 stem cells cultured in the presence of the ginsenosides together with dibutyryl cyclic AMP (dbcAMP) became parietal endoderm-like cells. Moreover, the expression of differentiation marker genes, such as laminin B1 and type IV collagen, was increased after treatment with the ginsenosides. Among the various purified ginsenosides, Rh, and Rh, were the most effective at causing differentiation of F9 cells. Since ginsenosides and glucocorticoid hormone have similar chemical structures, we examined the possibility of the involvement of a glucocorticoid receptor (GR) in the differentiation process induced by the ginsenosides. According to Southwestern blot analysis, a 94 kDa protein regarded as a GR was detected in F9 cells cultured in the medium containing the ginsenosides Rh-1 or Rh-2. In addition, F9 stem cells treated with the ginsenosides Rh-1 or Rh-2 and with RU486, a glucocorticoid antagonist with a high affinity for the GR, did not differentiate into endoderm cells morphologically, and the expression of laminin B1 gene was not induced in these cells. In a gel mobility shift assay, protein factors capable of binding to the glucocorticoid responsive element (GRE) specifically were detected in nuclear extracts of the ginsenoside-treated F9 cells. Moreover, overexpression of GR by cotransfection of GR expression vector and GRE-luciferase vector enhanced the transactivation activity of GRE promoter in the presence of ginsenosides Rh-1 or Rh-2 and was further augmented by dbcAMP. In addition, ginsenosides Rh-1 and Rh-2 bound to a GR assessed by whole-cell binding assay, even though the specific binding affinity was weaker compared to dexamethasone. Based on these data, we suggest that the ginsenosides Rh1 and Rh-2 cause the differentiation of F9 cells and the effects of ginsenosides might be exerted via binding with a GR or its analogous nuclear receptor. Copyright (C) 1996 Elsevier Science Ltd