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Methylation status of DJ-1 in leukocyte DNA of Parkinson's disease patients.

Tan Y, Wu L, Li D, Liu X, Ding J, Chen S - Transl Neurodegener (2016)

Bottom Line: It was found reduced in PD brains, CSF and saliva, although there were conflicting results.In SH-SY5Y cell model treated by methylation inhibitor, there was no significant change of DJ-1 expression in either mRNA level or protein level.Our results indicated that DNA methylation inhibitor didn't alter DJ-1 gene expression in SH-SY5Y cell model, and DNA methylation of DJ-1 promoter region in PBLs level might not be an efficient biomarker for PD patients.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology, and Institute of Neurology, Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, 200025 China.

ABSTRACT

Background: DJ-1 has been thought as a candidate biomarker for Parkinson's disease (PD). It was found reduced in PD brains, CSF and saliva, although there were conflicting results. How DJ-1 expression may be regulated is not clear. Recently, blood-based DNA methylation represents a highly promising biomarker for PD by regulating the causative gene expression. Thus, in this study, we try to explore whether blood-based DNA methylation of DJ-1 could be used as a biomarker to differentiate PD patients from normal control (NC), and whether DNA methylation could regulate DJ-1 expression in a SH-SY5Y cell model.

Methods: Forty PD patients and 40 NC were recruited in this study. DNA was extracted from peripheral blood leukocytes (PBLs). Methylation status of two CpG islands (CpG1 and CpG2) in promoter region of DJ-1 was explored by bisulfite specific PCR-based sequencing method. Methylation inhibitor 5-Aza-dC was used to treat SH-SY5Y cell line, DJ-1 level was detected in both mRNA and protein level.

Results: CpG sites in these two CpG islands (CpG1 and CpG2) of DJ-1 were unmethylated in both PD and NC group. In SH-SY5Y cell model treated by methylation inhibitor, there was no significant change of DJ-1 expression in either mRNA level or protein level.

Conclusions: Our results indicated that DNA methylation inhibitor didn't alter DJ-1 gene expression in SH-SY5Y cell model, and DNA methylation of DJ-1 promoter region in PBLs level might not be an efficient biomarker for PD patients.

No MeSH data available.


Related in: MedlinePlus

The mRNA and protein levels of DJ-1 did not change in 5-Aza-dC treated SH-SY5Y cells. Western blot and RT-PCR showed no significant change of DJ-1 protein level (a & b) and mRNA level (c) among 5-Aza-dC treated, DMSO treated and mock cells. Bar plot indicated the statistical analysis of DJ-1 protein level (b) and mRNA level (c) (Mean ± SD, *p < 0.05)
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Fig3: The mRNA and protein levels of DJ-1 did not change in 5-Aza-dC treated SH-SY5Y cells. Western blot and RT-PCR showed no significant change of DJ-1 protein level (a & b) and mRNA level (c) among 5-Aza-dC treated, DMSO treated and mock cells. Bar plot indicated the statistical analysis of DJ-1 protein level (b) and mRNA level (c) (Mean ± SD, *p < 0.05)

Mentions: Previous whole genome bisulfite sequencing (WGBS) study showed that DNA methylation can be detected in various regions, including CGIs, gene bodies and tandem repeating-containing regions [15]. Moreover, methylation levels in these regions can regulate the transcription levels [15, 16]. Although the CpG sites probed in this study were unmethylated in both PD and NC group, the other CpG sites out of the promoter region haven’t been determined. In order to test whether CpG methylation level can regulate the DJ-1 expression, methylation inhibitor 5-Aza-dC was used to demethylate the CpG sites in a SH-SY5Y cell model, then DJ-1 expression was detected in the mRNA and protein level. Our results showed DJ-1 expression didn’t have significant change in both the mRNA and protein level in SH-SY5Y cells treated with 2.5 mM or 5 mM 5-Aza-dC compared with the untreated cells (Fig. 3). Thus, the results indicated that DNA methylation had no regulatory effects on DJ-1 expression in SH-SY5Y cell model.Fig 3


Methylation status of DJ-1 in leukocyte DNA of Parkinson's disease patients.

Tan Y, Wu L, Li D, Liu X, Ding J, Chen S - Transl Neurodegener (2016)

The mRNA and protein levels of DJ-1 did not change in 5-Aza-dC treated SH-SY5Y cells. Western blot and RT-PCR showed no significant change of DJ-1 protein level (a & b) and mRNA level (c) among 5-Aza-dC treated, DMSO treated and mock cells. Bar plot indicated the statistical analysis of DJ-1 protein level (b) and mRNA level (c) (Mean ± SD, *p < 0.05)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
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getmorefigures.php?uid=PMC4815061&req=5

Fig3: The mRNA and protein levels of DJ-1 did not change in 5-Aza-dC treated SH-SY5Y cells. Western blot and RT-PCR showed no significant change of DJ-1 protein level (a & b) and mRNA level (c) among 5-Aza-dC treated, DMSO treated and mock cells. Bar plot indicated the statistical analysis of DJ-1 protein level (b) and mRNA level (c) (Mean ± SD, *p < 0.05)
Mentions: Previous whole genome bisulfite sequencing (WGBS) study showed that DNA methylation can be detected in various regions, including CGIs, gene bodies and tandem repeating-containing regions [15]. Moreover, methylation levels in these regions can regulate the transcription levels [15, 16]. Although the CpG sites probed in this study were unmethylated in both PD and NC group, the other CpG sites out of the promoter region haven’t been determined. In order to test whether CpG methylation level can regulate the DJ-1 expression, methylation inhibitor 5-Aza-dC was used to demethylate the CpG sites in a SH-SY5Y cell model, then DJ-1 expression was detected in the mRNA and protein level. Our results showed DJ-1 expression didn’t have significant change in both the mRNA and protein level in SH-SY5Y cells treated with 2.5 mM or 5 mM 5-Aza-dC compared with the untreated cells (Fig. 3). Thus, the results indicated that DNA methylation had no regulatory effects on DJ-1 expression in SH-SY5Y cell model.Fig 3

Bottom Line: It was found reduced in PD brains, CSF and saliva, although there were conflicting results.In SH-SY5Y cell model treated by methylation inhibitor, there was no significant change of DJ-1 expression in either mRNA level or protein level.Our results indicated that DNA methylation inhibitor didn't alter DJ-1 gene expression in SH-SY5Y cell model, and DNA methylation of DJ-1 promoter region in PBLs level might not be an efficient biomarker for PD patients.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology, and Institute of Neurology, Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, 200025 China.

ABSTRACT

Background: DJ-1 has been thought as a candidate biomarker for Parkinson's disease (PD). It was found reduced in PD brains, CSF and saliva, although there were conflicting results. How DJ-1 expression may be regulated is not clear. Recently, blood-based DNA methylation represents a highly promising biomarker for PD by regulating the causative gene expression. Thus, in this study, we try to explore whether blood-based DNA methylation of DJ-1 could be used as a biomarker to differentiate PD patients from normal control (NC), and whether DNA methylation could regulate DJ-1 expression in a SH-SY5Y cell model.

Methods: Forty PD patients and 40 NC were recruited in this study. DNA was extracted from peripheral blood leukocytes (PBLs). Methylation status of two CpG islands (CpG1 and CpG2) in promoter region of DJ-1 was explored by bisulfite specific PCR-based sequencing method. Methylation inhibitor 5-Aza-dC was used to treat SH-SY5Y cell line, DJ-1 level was detected in both mRNA and protein level.

Results: CpG sites in these two CpG islands (CpG1 and CpG2) of DJ-1 were unmethylated in both PD and NC group. In SH-SY5Y cell model treated by methylation inhibitor, there was no significant change of DJ-1 expression in either mRNA level or protein level.

Conclusions: Our results indicated that DNA methylation inhibitor didn't alter DJ-1 gene expression in SH-SY5Y cell model, and DNA methylation of DJ-1 promoter region in PBLs level might not be an efficient biomarker for PD patients.

No MeSH data available.


Related in: MedlinePlus