| Objective: To analyze the DNA methylation level in the promoter region of the susceptible gene and to compare the DNA methylation level in the promoter region of the case control group,select the sites of significant differences to further expand the sample verification,in order to find the epigenetic marker of AITD Provide a theoretical basis.Methods: Twenty-two inpatients in our department from June 2016 to December 2016 were selected according to the inclusion criteria of the case-control group,including 5 cases of control and 17 cases of GDO(including 10 cases of GD and 10 cases of HT).Data were collected and peripheral blood samples DNA extraction,bisulfite conversion,PCR amplification,in vitro transcription and other methylation quantitative detection of methylated significant sites were found to expand the sample size of the locus where the primer detection sequence further detection analysis,Control 35 cases,64 cases(including GD29 cases,HT35 cases)to detect the degree of methylation of the sequence.Result: Six sites with failed amplification and three sites with no methylation were completely removed,and 15 loci were obtained for analysis.A total of 5 sites were found in different groups of samples were significantly different(P<0.005),respectively,the fifth CTLA4 locus,CD40 eleventh locus,IL2 RA 21,22,23 were statistically(P<0.005).The number of CD40 was chosen as the eleventh region,and the expanded sample size was used to verify that the CD40 gene promoter The loci of 20 CG loci detected by the third locus in the region were significantly different between the case and control groups(P<0.005)Conclusion: There was a significant difference in the methylation status between the case control group and the hypomethylation level in the case group might be a susceptible methylation zone,which may be associated with autoimmune thyroid disease.It is necessary to further verify the methylation sites in the promoter region of the CD40 gene in different samples. |
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