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

doi:10.1016/j.neulet.2017.10.013

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 journal › Article › peer-review

7 Scopus citations

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 language	English (US)
Pages (from-to)	91-97
Number of pages	7
Journal	Neuroscience Letters
Volume	662
DOIs	https://doi.org/10.1016/j.neulet.2017.10.013
State	Published - Jan 1 2018
Externally published	Yes

Keywords

Cell-specific expression
DNA methylation
Epigenetics
Human dopamine transporter

ASJC Scopus subject areas

General Neuroscience

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10.1016/j.neulet.2017.10.013

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title = "The correlation between DNA methylation and transcriptional expression of human dopamine transporter in cell lines",

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.",

keywords = "Cell-specific expression, DNA methylation, Epigenetics, Human dopamine transporter",

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

note = "Publisher Copyright: {\textcopyright} 2017",

year = "2018",

month = jan,

day = "1",

doi = "10.1016/j.neulet.2017.10.013",

language = "English (US)",

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T1 - The correlation between DNA methylation and transcriptional expression of human dopamine transporter in cell lines

AU - Zhai, Desheng

AU - Li, Songji

AU - Dong, Gaopan

AU - Zhou, Dushuang

AU - Yang, Yuxin

AU - Wang, Xin

AU - Zhao, Ying

AU - Yang, Yunlei

AU - Lin, Zhicheng

PY - 2018/1/1

Y1 - 2018/1/1

N2 - 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.

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.

KW - Cell-specific expression

KW - DNA methylation

KW - Epigenetics

KW - Human dopamine transporter

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SN - 0304-3940

VL - 662

SP - 91

EP - 97

JO - Neuroscience Letters

JF - Neuroscience Letters

ER -

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

Abstract

Keywords

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