2024-03-28T23:01:58Z
https://nagoya.repo.nii.ac.jp/oai
oai:nagoya.repo.nii.ac.jp:00012550
2023-01-16T03:59:15Z
499:960:961
ATRA inhibits ceramide kinase transcription through an ATRA-related transcription factor, COUP-TFI, in a human neuroblastoma cell line, SH-SY5Y
ヒト神経芽細胞腫細胞株SH-SY5Y細胞のATRA誘導性転写因子COUP-TFIによるセラミドキナーゼ遺伝子の発現抑制
村上, 真史
MURAKAMI, Masashi
open access
All-trans retinoic acid
neuronal differentiation
SH-SY5Y cell
ceramide kinase
promoter analysis
COUP-TFI
Ceramide is the central lipid in the sphingolipid metabolism. Ceramide kinase (CERK) and its product, ceramide 1-phosphate (C1P), have been implicated in various cellular functions. However, the regulatory mechanism of CERK gene expression remains to be determined. Here, we examined CERK mRNA level during all-trans retinoic acid (ATRA)-induced differentiation of a human neuroblastoma cell line, SH-SY5Y. ATRA reduced CERK mRNA and protein levels. Overexpression and siRNA of CERK revealed that CERK is inhibitory against ATRA-induced neuronal differentiation and cell growth arrest. ATRA inhibited the transcriptional activity of 5’-promoter of CERK. Truncation and mutation study suggests that ATRA-responsible region was mainly located in the tandem retinoic acid responsive elements (RARE) between -40 bp and the first exon. The electrophoresis mobility shift assay revealed that ATRA produced two retarded bands, which were erased by antibody against COUP-TFI, RARα and RXRα, respectively. DNA pull-down assay confirmed increased binding of these transcription factors to RARE. Transient expression of RAR, RXR and COUP-TFI and siRNA transfection of these genes revealed that COUP-TFI inhibited CERK mRNA. Furthermore, chromatin immunoprecipitation assay showed the recruitment of co-repressors as well as three transcription factors. These results suggest that COUP-TFI was the ATRA-responsive suppressive transcription factor of CERK gene transcription.
名古屋大学博士学位論文 学位の種類:博士(医療技術学) (課程) 学位授与年月日 平成22年3月25日
2010-03-25
eng
doctoral thesis
http://hdl.handle.net/2237/14433
https://nagoya.repo.nii.ac.jp/records/12550
甲第8661号
博士(医療技術学)
2010-03-25
13901
名古屋大学
NAGOYA University
https://nagoya.repo.nii.ac.jp/record/12550/files/k8661.pdf
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2018-02-20