Department of Internal Medicine, Charles Drew University of Medicine & Science, Los Angeles, CA 90059, USA.
J Steroid Biochem Mol Biol. 2010 Mar;119(1-2):73-83. doi: 10.1016/j.jsbmb.2010.01.001. Epub 2010 Jan 11.
The vitamin D receptor (VDR) and its ligand 1,25D play an important role in regulating cell growth and cell fate. We examined the effect of 1,25D on cell morphology, cell proliferation, cell cycle progression and apoptosis on mesenchymal multipotent cells. Multipotent cells were treated with and without 1,25D in a time- and dose-dependent manner. Changes in cell morphology were evaluated by a green fluorescence fluorocrome. Cell proliferation was determined by the Formazan assay and PCNA antigen expression. The expression of genes related to the cell cycle was analyzed by DNA microarrays, RT(2)PCR arrays and western blots. Apoptosis was evaluated by TUNEL assay, and the expression of pro- and anti-apoptotic related genes by RT(2)PCR arrays and western blots. 1,25D inhibited cell proliferation, induced cell cycle arrest, and promoted accumulation of cells in G0/G1 phase without inducing apoptosis. An increase in cell size was associated with a decrease in the GTPase Rho and the atypical Rho family GTPase Rhou/Wrch-1 expression without inducing Wnt-1 expression. Survivin expression was also increased and may represent a novel 1,25D-mediated pathway regulating tissue injury and fibrosis. The data provide a mechanistic explanation for the anti-proliferative and anti-apoptotic properties of 1,25D in mesenchymal multipotent cells.
维生素 D 受体 (VDR) 及其配体 1,25D 在调节细胞生长和细胞命运方面发挥着重要作用。我们研究了 1,25D 对间充质多能细胞的细胞形态、细胞增殖、细胞周期进程和细胞凋亡的影响。多能细胞以时间和剂量依赖的方式用和不用 1,25D 进行处理。通过绿色荧光荧光染料评估细胞形态的变化。通过 Formazan 测定和 PCNA 抗原表达来确定细胞增殖。通过 DNA 微阵列、RT(2)PCR 阵列和 Western blot 分析与细胞周期相关的基因表达。通过 TUNEL 测定评估细胞凋亡，并通过 RT(2)PCR 阵列和 Western blot 分析促凋亡和抗凋亡相关基因的表达。1,25D 抑制细胞增殖，诱导细胞周期停滞，并促进细胞在 G0/G1 期积累，而不诱导细胞凋亡。细胞体积的增加与 GTPase Rho 和非典型 Rho 家族 GTPase Rhou/Wrch-1 的表达减少有关，而不会诱导 Wnt-1 的表达。Survivin 的表达也增加，可能代表一种新的 1,25D 介导的途径，调节组织损伤和纤维化。该数据为 1,25D 在间充质多能细胞中的抗增殖和抗凋亡特性提供了机制解释。
