Expression Of Clock Genes Per1,Per2 And Per3 In Patients With Head And Neck Squamous Cell Carcinoma And Its Correlation With Clinical Factors

To investigate the expression of Per1,Per2 and Per3 in normal issues of head and neck squamous cell carcinoma in terms of pathology,protein and gene level expression;detect the relationship between Perl,Per2 and Per3 gene expression level and clinical stages of head and neck squamous cell carcinoma;discuss the association of Perl,Per2 and Per3 protein positive expression and clinical features,tumor clinical stages and survival of head and neck squamous cell carcinoma for the purpose of offering reference indicators for clinical assessment of patient prognosis.Method:27 biopsy samples were colleted,including 7 normal tissue samples of head and neck and 20 tissue samples in head and neck squamous cell carcinoma,were included.Real-time fluorescent quantitative PCR method was used to detect the expression level of Per1,Per2 and Per3 genes in head and neck squamous cell carcinoma and head and neck normal tissues,and relative quantitative method was used to analyze the expression difference of Perl,Per2 and Per3 genes in head and neck squamous cell carcinoma and head and neck normal tissue.Based on patient clinical stages,(Ⅰ-Ⅱ)was identified as early group and(III-IV)was identified as late group and the association of Perl,Per2 and Per3 gene expression differences with head and neck squamous cell carcinoma patients in early and late stages.In the same time,80 paraffin blocks with available clinical treatment data were collected in Guizhou Cancer Hospital,including 60 paraffin blocks of head and neck squamous cell carcinoma tissue,and 20 paraffin blocks of head and neck normal tissue.Immunohistochemistry was used to detect Perl,Per2 and Per3 protein expression,the product of multiplying by staining grading and staining cell percentage was used to determine Perl,Per2 and Per3 protein expression level,which was used to analyze the relationship between Per1,Per2 and Per3 protein and head and neck squamous cell carcinoma tissue and head and neck normal tissue,and the protein level was used to analyze its association with patient clinical features,carcinoma clinical stages and survival.Result:Based on gene level detection,expression of Perl,Per2 and Per3 genes was found in head and neck squamous cell carcinoma tissue and head and neck normal tissue.For the samples of head and neck squamous cell carcinoma tissue,Per1,Per2 and Per3 gene expression was reduced compared with normal tissue(P<0.01).In patients in late stage,reduction levels of Perl,Per2 and Per3 genes was higher than patients in early stage,but only reduction levels of Per2 and Per3 genes were of statistical significance.According to immunohistochemistry test,protein positive expression rate of Perl,Per2 and Per3 genes in normal tissue was higher that of Perl,Per2 and Per3 genes in head and neck squamous cell carcinoma tissue(P<0.05),and the protein expression level of the former was related to carcinoma clinical stages(P<0.05)and unrelated to patient age and sex.Head and neck squamous cell carcinoma in patients with Perl,Per2,Per3 protein positive group were 1,2,3 years of OS was significantly higher than the negative group,the difference was statistically significant(P<0.05);Perl,Per2,Per3 protein positive group were 1,2,3 years of PFS and RFS was significantly higher than the negative group,the difference was statistically significant(P<0.05).Conclusion:Reduction in Perl,Per2 and Per3 gene expression may be correlated to the occurrence of head and neck squamous cell carcinoma,and reduction extent of Per2 and Per3 genes in patients in late stage is higher than that in patients of head and neck squamous cell carcinoma in early stage;compared with patients with negative Per1、Per3 protein expression,patients with positive Per1 and Per3 protein expression may have longer OS and show the trend of extended PFS and RFS,which may indicate that Perl,Per2 and Per3 genes may play a critical role in the occurrence and development of head and neck squamous cell carcinoma.