CpG Array Analysis of Histone H3 Lysine 4 Trimethylation by Chromatin Immunoprecipitation Linked to Microarrays Analysis in Peripheral Blood Mononuclear Cells of IgA Nephropathy Patients
Yonsei Medical Journal
; : 377-385, 2012.
Article
在 En
| WPRIM
| ID: wpr-154804
Responsible library:
WPRO
ABSTRACT
PURPOSE:
The purpose of the present study was to investigate the aberrance of histone H3 lysine 4 trimethylation (H3K4me3) in patients with IgA Nephropathy (IgAN). MATERIALS ANDMETHODS:
In this study, H3K4me3 variations in peripheral blood mononuclear cells (PBMCs) from 15 IgAN patients and 15 healthy subjects were analyzed using chromatin immunoprecipitation linked to microarrays analysis (ChIP-chip). ChIP real-time PCR was used to validate the microarray results. Expression analysis by quantitative real-time PCR (qRT-PCR) revealed correlations between mRNA and H3K4me3 levels. DNA methylation status was analyzed by quantitative methylation-specific PCR.RESULTS:
We found that 321 probes displayed significant H3K4me3 differences in IgAN patients compared with healthy controls. Among these probes, 154 probes displayed increased H3K4me3 and 167 probes demonstrated decreased H3K4me3. For further validation, we selected 4 key relevant genes (FCRL4, GALK2, PTPRN2 and IL1RAPL1) to study. The results of ChIP real-time PCR coincided well with the microarray data. Quantitative RT-PCR revealed the correlations between the mRNA expression and the methylation levels of H3K4me3. Different degrees of DNA methylation alterations appeared on the selected positive genes.CONCLUSION:
Our studies indicated that there were significant alterations in H3K4me3 in IgAN patients. These findings may help to explain the disturbed immunity and abnormal glycosylation involved in IgAN patients.Key words
全文:
1
索引:
WPRIM
主要主题:
Leukocytes, Mononuclear
/
Histones
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Case-Control Studies
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Oligonucleotide Array Sequence Analysis
/
Chromatin Immunoprecipitation
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Real-Time Polymerase Chain Reaction
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Glomerulonephritis, IGA
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Lysine
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Methylation
研究类型:
Observational_studies
限制:
Adult
/
Female
/
Humans
/
Male
语言:
En
期刊:
Yonsei Medical Journal
年:
2012
类型:
Article