The Methylation Of PAX1 Gene And Polymorphisms Of Folate Metabolic Genes In Oral Squamous Cell Carcinoma

Objective:To identify novel diagnosis and prognosis biomarkers and clinic treatment targets for oral squamous cell carcinoma(OSCC)we investigated the methylation status of Paired Box1 gene(PAX1)in OSCC tissues,their adjacent mucosa and surgical margin.We characterized the genotype of methylenetetrahydrofolate reductase(MTHFR)and methioninesynthase reductase(MTRR)genes in oral cancer tissues.Finally,we investigated the relationship between methylation status of PAX1 gene and the polymorphisms of folate metabolic genes including MTHFR gene C677 T and A1298 C,and MTRR gene A66 G in OSCC cases in Xiangtan.Methods:The specimens for this study came from patients in the First People's Hospital of Xiangtan City and the Central Hospital of Xiangtan City.ALL subjects were genetically unrelated ethnic Han Chinese from Xiangtan city.Cases newly histopathologically diagnosed as incident oral squamous cell carcinoma were consequently recruited from the First People's Hospital of Xiangtan City and the CentralHospital of Xiangtan City.The specimens included fresh frozen OSCC tissues(n=30),their adjacent mucosa tissues(n=30)and surgical margin tissues(n=30).The methylation status of PAX1 gene was determined by quantitative methylation specific PCR.The MTHFR(C677T,A1298C)and MTRR(A66G)gene polymorphisms were determined using the Pyrosequencing method.Take the Fisher's exact probability statistical analysis: OSCC tissues and adjacent mucosa tissues,surgical margin tissues,with the adjacent mucosa tissues and surgical margin tissues PAX1 gene methylation rate if there is a difference.With Odds ratio(OR)and95% confidence interval(CI)analysis of 30 cases of OSCC tissues PAX1 gene methylation whether there is a connection between the polymorphism of MTHFR 677(C?T)and 1298(A?C)and MTRR 66(A ?G).Result:From 30 cases of OSCC tissues,30 cases of adjacent mucosa tissues and 30 cases of surgical margin tissues,hypermethylation of the PAX1 gene was found in 22 cases of OSCC tissues(73.3%),13 cases of adjacent mucosa tissues(43.3%),and 5 cases of surgical margin tissues(16.7%),respectively.The methylation of PAX1 gene was significantly higher in OSCC tissues,and the differences between OSCC tissues and their adjacent mucosa and surgical margin tissues were statistically significant(P<0.05).In 30 cases of OSCC tissues,MTHFR 677(C?T)genotype of CC was identified in 15 cases.Genotype of CT was identified in 13 cases.Genotype of TT was identified in 2 cases.MTHFR1298(A?C)genotype of AA was identified in 18 cases.Genotype of AC was identified in 12 cases.Genotype of CC was not identified.MTRR 66(A?G)genotype of AA was found in 13 cases.Genotype of AG was found in 16 cases.Genotype of GG was found in 1 cases.For the MTHFR C677 T,A1298C,and the MTRR A66 G gene polymorphisms,the difference was not statistically significant between the unmethylated and hypermethylated PAX1 cases.Conclusion:1.The PAX1 gene is hypermethylated in the OSCC and adjacent mucosa tissues.The PAX1 gene hypermethylation plays a crucial role in the diagnosis of OSCC.2.There is no connection between the methylation of PAX1 gene and the polymorphisms of MTHFR(C677T,A1298C)and MTRR(A66G)in OSCC tissues.