Analysis on A(TA)n TAA polymorphism of UGT1A1 gene promoter by fluorescence real-time quantitative PCR / 国际检验医学杂志

Objective To develop a new method to detect A (TA)n TAA polymorphism in the UGT1A1 gene promoter by fluo‐rescence real‐time quantitative PCR (RQ‐PCR) .Methods Genomic DNA was extracted from peripheral blood in 16 patients with Gilbert′s syndrome and 66 healthy individuals .The polymorphic A(TA)n TAA sequence in the UGT1A1 gene promoter was deter‐mined by DNA sequencing .A pair of primers and two TaqMan probes labeled with either 5′FAM or VIC reporter dye incorporated a 3′minor groove binder were designed .The A(TA)n TAA polymorphisms in the UGT1A1 gene promoter were identified by RQ‐PCR for all research subjects .The sensitivity and specificity of RQ‐PCR for detecting the A(TA)nTAA polymorphisms were veri‐fied by DNA sequencing method .Results The homozygous A(TA)7TAA polymorphism was found in the promoter region of the UGT1A1 gene in all 16 patients with Gilbert′s syndrome by using RQ‐PCR .The homozygous A(TA)6TAA polymorphism was foundin46healthysubjects,whiletheheterozygousA(TA)6TAA/A(TA)7TAApolymorphismwasfoundinother20healthysub‐jects .All A(TA)nTAA polymorphisms in the promoter region of the UGT1A1 gene identified by RQ‐PCR were consistent with that of DNA sequencing .Conclusion It is a sensitive ,specific and simple method to detect the A (TA)n TAA polymorphisms in the promoter region of the UGT1A1 gene by RQ‐PCR ,which can be promoted and applied in clinic .

Methylation of SYK Gene Promoter Region in Nasopharyngeal Carcinoma / 现代检验医学杂志

Objective To detect methylation of spleen tyrosine kinase (SYK)gene promoter region in nasopharyngeal carcino-ma,and to explore relationship between carcinogenesis of nasopharyngeal carcinoma and methylation of SYK gene promoter region.Methods A total of 52 patients with nasopharyngeal carcinoma and 26 patients with chronic rhinitis from Baoan People’s Hospital of Shenzhen Hospital of Peking University were enrolled in this study between February 2012 and August 2014.All cases were diagnosed by pathological examination.Methylation specific-polymerase chain reaction assay was per-formed to detect methylation status of SYK gene promoter region,the rate of methylation was compared for patients with nasopharyngeal carcinoma and patients with chronic rhinitis.Results The methylation of promoter region of SYK gene was detected for 11 biopsy samples among 52 nasopharyngeal carcinoma patients,the methylation frequency was 21.2% in naso-pharyngeal carcinoma biopsy samples,while methylation was not found in 26 chronic rhinitis biopsy samples.Conclusion Low methylation of promoter region of SYK gene was found in nasopharyngeal carcinoma.It suggests that methylation of SYK gene promoter region may contribute to carcinogenesis of nasopharyngeal carcinoma.