Analysis of the vitamin D system in basal cell carcinomas (BCCs)
Analysis of the vitamin D system in basal cell carcinomas (BCCs)
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DOI：
10.1038/labinvest.3700096
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发表时间：
2004-06-01
影响因子：
5
通讯作者：
Reichrath, J
中科院分区：
文献类型：
--
作者：
Mitschele, T;Diesel, B;Reichrath, J
使用实时PCR（LightCycler）和免疫组化，我们分析了维生素D系统的关键成分在基底细胞癌（BCC）和正常人皮肤（NS）中的表达。使用链霉抗生物素蛋白-过氧化物酶技术证明了基底细胞癌中VDR免疫反应性增加。检测维生素D受体（VDR）和骨化三醇合成代谢的主要酶（维生素D-25-羟化酶[25-OHase]、25-羟维生素D-1 α-羟化酶[1alpha-OHase]、1，25-二羟维生素D-24-羟化酶[24-OHase]）在基底细胞癌和NS中的RNA表达。表达水平以靶基因（VDR、1 α-OHase、25-OHase、24-OHase）和管家基因甘油醛-3-磷酸脱氢酶（GAPDH）（作为内部对照）之间的比率确定。VDR/GAPDH（BCC：16.54; NS：0.00021）、1 α-OHase/GAPDH（BCC：0.739; NS 0.000803）和24-OHase/GAPDH（BCC：0.00585; NS 0.00000366）的mRNA比值中位数在BCC中显著升高（Wilcoxon-Mann-Whitney U检验）。与此相反，25-OHase/GAPDH的mRNA比值中位数（BCC：0.17; NS：0.016）在BCC中与NS相比没有显著改变。此外，我们首次报道了1 α-OHase剪接变异体在基底细胞癌和NS中的表达，并使用传统的RT-PCR检测。总之，我们的研究结果为维生素D代谢物的内源性合成和代谢以及VDR表达可能调节基底细胞癌的生长特性的概念提供了支持性证据。新的维生素D类似物，几乎没有钙的副作用，他们的前体，或抑制剂的24-羟化酶可能提供一个新的方法来预防或治疗基底细胞癌。我们在此首次描述的1 α-羟化酶的替代转录物在基底细胞癌和NS中的功能及其对活性水平的影响，将在未来的实验中进行研究。
Using real-time PCR (LightCycler) and immunohistochemistry, we have analyzed expression of key components of the vitamin D system in basal cell carcinomas (BCCs) and normal human skin (NS). Increased VDR-immunoreactivity was demonstrated in BCCs using a streptavidin-peroxidase technique. RNA expression of vitamin D receptor (VDR) and of main enzymes involved in synthesis and metabolism of calcitriol (vitamin D-25-hydroxylase [25-OHase], 25-hydroxyvitamin D-1alpha-hydroxylase [1alpha-OHase], 1,25-dihydroxyvitamin D-24-hydroxylase [24-OHase]) was detected in BCCs and NS. Expression levels were determined as ratios between target genes (VDR, 1alpha-OHase, 25-OHase, 24-OHase) and the housekeeping gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as internal control. Median of mRNA ratios for VDR/GAPDH (BCCs: 16.54; NS: 0.00021), 1alpha-OHase/GAPDH (BCCs: 0.739; NS 0.000803) and 24-OHase/GAPDH (BCCs: 0.00585; NS 0.000000366) was significantly (Wilcoxon-Mann-Whitney U-test) elevated in BCCs. In contrast, median of mRNA ratio for 25-OHase/GAPDH (BCCs: 0.17; NS: 0.016) was not significantly altered in BCCs as compared to NS. Additionally, we report for the first time expression of 1alpha-OHase splice variants in BCCs and NS, that were detected using conventional RT-PCR. In conclusion, our findings provide supportive evidence for the concept that endogeneous synthesis and metabolism of vitamin D metabolites as well as VDR expression may regulate growth characteristics of BCCs. New vitamin D analogs that exert little calcemic side effects, their precursors, or inhibitors of 24-OHase may offer a new approach for the prevention or therapy of BCCs. The function of alternative transcripts of 1alpha-OHase that we describe here for the first time in BCCs and NS and their effect on activity level has to be investigated in future experiments.