The correlation between DNA methylation and transcriptional expression of human dopamine transporter in cell lines

Desheng Zhai, Songji Li, Gaopan Dong, Dushuang Zhou, Yuxin Yang, Xin Wang, Ying Zhao, Yunlei Yang, Zhicheng Lin

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

This study aims to investigate the relationship between DNA methylation and expression of human dopamine transporter (hDAT). We examined methylation status of hDAT in cells with various hDAT expression levels, including two dopaminergic neural cell lines (SK-N-AS and SH-SY-5Y) and one non-dopaminergic cell line (HEK293) by bisulfite sequencing PCR(BSP). The effects of DNA methyltransferase inhibitor 5-aza-dC or/and histone deacetylase inhibitor (HDACi, sodium butyrate, NaB) on the DNA methylation status and mRNA expression levels of hDAT were examined. The results revealed marked hypomethylation of the two promoter regions (−1214 to −856 bp and −48 to 439 bp, the first base of exon 1 was taken as +1 bp)of hDAT in SK-N-AS (4.7% ± 2.0 mC and 3.5% ± 1.0 mC, respectively) compared with SH-SY-5Y (88.0% ± 4.4%mC and 81.1% ± 8.8%mC) and HEK293 (90.7% ± 2.4 mC and 84.4% ± 8.6% mC) cell lines, indicating a cell-specific methylation regulation of hDAT. 5-aza-dC and NaB decreased hypermethylation,while increase hDAT expression in SH-SY-5Y cells and recovered hDAT mRNA expression in HEK293 cells. DNA methylation enabled the cell-specific differential expression of the hDAT gene. hDAT silencing was reversed by the introduction of DNA hypomethylation via 5-aza-dC or/and NaB.

Original languageEnglish (US)
Pages (from-to)91-97
Number of pages7
JournalNeuroscience Letters
Volume662
DOIs
StatePublished - Jan 1 2018
Externally publishedYes

Fingerprint

Dopamine Plasma Membrane Transport Proteins
DNA Methylation
Cell Line
Methylation
Messenger RNA
Histone Deacetylase Inhibitors
Butyric Acid
HEK293 Cells
DNA
Methyltransferases
Genetic Promoter Regions
Exons
Polymerase Chain Reaction

Keywords

  • Cell-specific expression
  • DNA methylation
  • Epigenetics
  • Human dopamine transporter

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

The correlation between DNA methylation and transcriptional expression of human dopamine transporter in cell lines. / Zhai, Desheng; Li, Songji; Dong, Gaopan; Zhou, Dushuang; Yang, Yuxin; Wang, Xin; Zhao, Ying; Yang, Yunlei; Lin, Zhicheng.

In: Neuroscience Letters, Vol. 662, 01.01.2018, p. 91-97.

Research output: Contribution to journalArticle

Zhai, Desheng ; Li, Songji ; Dong, Gaopan ; Zhou, Dushuang ; Yang, Yuxin ; Wang, Xin ; Zhao, Ying ; Yang, Yunlei ; Lin, Zhicheng. / The correlation between DNA methylation and transcriptional expression of human dopamine transporter in cell lines. In: Neuroscience Letters. 2018 ; Vol. 662. pp. 91-97.
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AU - Zhao, Ying

AU - Yang, Yunlei

AU - Lin, Zhicheng

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AB - This study aims to investigate the relationship between DNA methylation and expression of human dopamine transporter (hDAT). We examined methylation status of hDAT in cells with various hDAT expression levels, including two dopaminergic neural cell lines (SK-N-AS and SH-SY-5Y) and one non-dopaminergic cell line (HEK293) by bisulfite sequencing PCR(BSP). The effects of DNA methyltransferase inhibitor 5-aza-dC or/and histone deacetylase inhibitor (HDACi, sodium butyrate, NaB) on the DNA methylation status and mRNA expression levels of hDAT were examined. The results revealed marked hypomethylation of the two promoter regions (−1214 to −856 bp and −48 to 439 bp, the first base of exon 1 was taken as +1 bp)of hDAT in SK-N-AS (4.7% ± 2.0 mC and 3.5% ± 1.0 mC, respectively) compared with SH-SY-5Y (88.0% ± 4.4%mC and 81.1% ± 8.8%mC) and HEK293 (90.7% ± 2.4 mC and 84.4% ± 8.6% mC) cell lines, indicating a cell-specific methylation regulation of hDAT. 5-aza-dC and NaB decreased hypermethylation,while increase hDAT expression in SH-SY-5Y cells and recovered hDAT mRNA expression in HEK293 cells. DNA methylation enabled the cell-specific differential expression of the hDAT gene. hDAT silencing was reversed by the introduction of DNA hypomethylation via 5-aza-dC or/and NaB.

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