TY - JOUR
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
N1 - Funding Information:
This work was supported by National Natural Science Fundation of China 81100956 (to YZ). A Innovation Project Foundation of He’nan Educational Committee, China 14HASTIT034 (to YZ), Foundation for University Key Teacher by He’nan Educational Committee, China , 2011 (to DZ), Key project of science and technology research by He’nan Educational Committee, China , 13A310859 and 14A330005 (to DZ), Undergraduate Innovation Project by Ministry of Educational of China ( 201310472003 ; 201310472011 ; 201310472043 ), Support Project for the Disciplinary Group of Psychology and Neuroscience, Xinxiang Medical University 2016PN-KFKT-30 (to YZ), Foundation of Henan Key Laboratory of Neural Regeneration HNSJXF-2016-008 (to YZ)
Publisher Copyright:
© 2017
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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U2 - 10.1016/j.neulet.2017.10.013
DO - 10.1016/j.neulet.2017.10.013
M3 - Article
C2 - 29030220
AN - SCOPUS:85031101935
SN - 0304-3940
VL - 662
SP - 91
EP - 97
JO - Neuroscience Letters
JF - Neuroscience Letters
ER -