Screening for epigenetically masked genes in Kashin-Beck disease by microarray / 中华地方病学杂志

ABSTRACT

Objective We used the DNA methylation microarrays to investigate the differential methylation genes and loci sites in Kashin-Beck disease (KBD),to study the relationship between DNA methylation and KBD pathogenesis.Methods Totally 12 KBD adults and 12 healthy adults were selected and peripheral blood samples were collected and DNA was extracted.Illumina 450K bead-chip was applied to detect methylation status in KBD and healthy controls.Aberrant hyper-methylated sites were filtrated according to the P value after correction and methylation differences,together with GenomeStudio soft.Screened genes were validated using bisulfite sequencing polymerase chain reaction (BSP) technology.Results A total of 484 948 loci sites were analyzed and compared,93 differential methylated loci were found by comparing KBD and normal people,including 34 hypermethylated sites and 59 hypomethylated sites.There were 50 genes corresponding to the loci,43 genes not reported in literature.According to gene ontology analysis,the genes were involved in the immune response,antigen processing,phosphate and phosphoric acid metabolism and phosphorylation and the process of metal ions in combination.However,in the verification test using BSP method,there was no significant difference in methylation rate in human leukocyte antigen (HLA)-DRB1 between the case and the control group (48％ vs 70％,x2 =3.688,P ＞ 0.05).Conclusions The high and low differentially methylated sites in peripheral blood DNA of KBD patients are significantly different from those of the health control.HLA-DRB1 locus is not significantly different between the BSP verification test and methylation chip.