Abstract | PURPOSE: EXPERIMENTAL DESIGN: We did nested touchdown reverse transcription-PCR (RT-PCR) to identify CYP27B1 splice variants and real-time RT-PCR to quantify the expression of CYP27B1. A cell line was treated with calcitriol to determine the effect on the expression of CYP27B1, 1alpha,25-dihydroxyvitamin D(3)-24-hydroxylase (CYP24), and vitamin D(3) receptor (VDR). We generated three antibodies for the specific detection of CYP27B1 and splice variants. High-performance TLC was done to determine the endogenous CYP27B1 activity and the functionality of CYP27B1 splice variants. Using WST-1 assay, we determined the effect of vitamin D(3) metabolites on proliferation. RESULTS: CONCLUSIONS:
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Authors | Britta Diesel, Jens Radermacher, Matthias Bureik, Rita Bernhardt, Markus Seifert, Jörg Reichrath, Ulrike Fischer, Eckart Meese |
Journal | Clinical cancer research : an official journal of the American Association for Cancer Research
(Clin Cancer Res)
Vol. 11
Issue 15
Pg. 5370-80
(Aug 01 2005)
ISSN: 1078-0432 [Print] United States |
PMID | 16061850
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- Calcium Channel Agonists
- DNA, Complementary
- RNA, Messenger
- Receptors, Calcitriol
- Recombinant Proteins
- Cholecalciferol
- RNA
- Steroid Hydroxylases
- Vitamin D3 24-Hydroxylase
- 25-Hydroxyvitamin D3 1-alpha-Hydroxylase
- Calcitriol
- Calcifediol
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Topics |
- 25-Hydroxyvitamin D3 1-alpha-Hydroxylase
(genetics, metabolism)
- Alternative Splicing
- Animals
- Blotting, Western
- Brain Neoplasms
(metabolism)
- COS Cells
- Calcifediol
(metabolism)
- Calcitriol
(pharmacology)
- Calcium Channel Agonists
(pharmacology)
- Cell Line, Tumor
- Cell Proliferation
- Cholecalciferol
(metabolism)
- Chromatography, Thin Layer
- Cloning, Molecular
- DNA, Complementary
(metabolism)
- Gene Expression Regulation, Neoplastic
- Glioblastoma
(metabolism)
- Humans
- Plasmids
(metabolism)
- RNA
(metabolism)
- RNA, Messenger
(metabolism)
- Rats
- Receptors, Calcitriol
(metabolism)
- Recombinant Proteins
(chemistry)
- Reverse Transcriptase Polymerase Chain Reaction
- Steroid Hydroxylases
(metabolism)
- Transfection
- Vitamin D3 24-Hydroxylase
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