HaCaT cell line as a model system for vitamin D-3 metabolism in human skin
HaCaT cell line as a model system for vitamin D-3 metabolism in human skin
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DOI：
10.1111/1523-1747.ep12285640
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发表时间：
1997-01-01
影响因子：
6.5
通讯作者：
Lehmann, B
中科院分区：
文献类型：
--
作者：
Lehmann, B
以HaCaT细胞系为细胞培养模型，研究了骨化三醇(1,25（OH)(2)D-3）的合成和分解代谢。我们的研究结果表明，当骨化二醇（h -3- 25ohd(3)）作为底物时，表皮生长因子（EGF）或转化生长因子- α (tgf - α)在到达汇合前16小时内刺激HaCaT细胞，可增加骨化三醇的产生。EGF-和tgf - α诱导的（0.1 ~ 10 nM） H-3-25OHD(3)的1-羟基化是浓度依赖性的，但表现出不同的动力学，EGF诱导的骨化三醇合成与细胞融合程度成反比。EGF刺激对放线菌素D和环己亚胺敏感。与所使用的生长因子无关，与骨化三醇的生成量相比，H-3-24R,25(OH)(2)D-3的生成量和h -3-1,25(OH)(2)D-3到h -3-1,24,25(OH)(3)D-3的分解代谢出乎意料地低（小于或等于5%，小于或等于2%）。外源添加未标记的1,25(OH)(2) d - 3,1,24r (OH)(2)D-3、钙化三醇或24R，25(OH)(2)D-3，浓度低至10(-11)M，可有效抑制h -3-1,25(OH)(2)D-3的产生。这些结果表明，egf处理的HaCaT角化细胞可以为进一步研究维生素D-3途径及其与增殖和分化的关系提供服务，但必须考虑与培养的原代角化细胞或其他细胞系在骨化三醇合成和分解代谢方面的差异。
Synthesis and catabolism of calcitriol (1,25(OH)(2)D-3) were studied using HaCaT cell line as a cell culture model, Our results indicate that stimulation of HaCaT cells with epidermal growth factor (EGF) or transforming growth factor-alpha (TGF-alpha) within 16 h just prior to reaching confluence amplified the production of calcitriol when calcidiol (H-3-25OHD(3)) was used as a substrate. EGF- and TGF-alpha-induced (0.1-10 nM) 1-hydroxylation of H-3-25OHD(3) was concentration-dependent but showed different kinetics, Synthesis of calcitriol induced by EGF was inversely related to the degree of cellular confluence. Stimulation by EGF was an actinomycin D- and cycloheximide-sensitive process. Independently of the growth factor used, the production of H-3-24R,25(OH)(2)D-3 and the catabolism of H-3-1,25(OH)(2)D-3 to H-3-1,24,25(OH)(3)D-3 were unexpectedly low (less than or equal to 5% and less than or equal to 2%, as compared to the amount of calcitriol generated. Exogenous addition of unlabeled 1,25(OH)(2)D-3, 1,24R(OH)(2)D-3, calcipotriol, or 24R,25(OH)(2)D-3 at concentrations as low as 10(-11) M, potently inhibited the H-3-1,25(OH)(2)D-3 production. These results suggest that EGF-treated HaCaT keratinocytes could serve for further studies of the vitamin D-3 pathway and its relationship to proliferation and differentiation, but differences in calcitriol synthesis and catabolism from those in cultured primary keratinocytes or other cell Lines must be considered.