Clinical Role Of CD39 Expression In Head And Neck Squamous Cell Carcinoma And Its Related Impact Factors

Objective: To explore the association of CD39 expression level with clinicopathological features and prognosis of patients with HNSCC,assess the relationship of CD39 expression level to PD-L1,PD-L2,and ALDH1A1,as well as the subsets of tumor-infiltrating lymphocyte,and reveal the potential significance of CD39 in the pathogenesis/progression of HNSCC.Methods: Firstly,the expression level of CD39 mRNA between in buccal mucosa(BM) tissues and HNSCC tissues was compared via accessing Oncomine database.Immunohistochemistry(IHC)was to detect the expression level of CD39 protein in 85 HNSCC tissues,Spearman’s correlation analysis and multivariate logistic regression analysis were performed to screen the independent factors acting on the expression of CD39 protein in HNSCC tissues,and Kaplan-Meier curves was drawn and a multivariate Cox risk proportional regression model was constructed to evaluate the relationship between the expression level of CD39 protein and the prognosis of patients with HNSCC.IHC was performed once again in this study to detect the expression levels of PD-L1,PD-L2 and ALDH1A1 proteins in HNSCC tissues,as well as the distribution of CD3,CD8,CD103 and Foxp3 positive immunocytes,and further analyze the association between the expression level of CD39 in HNSCC tissues and the levels of these factors detected,and on the basis of factor analysis method,a multivariate Cox regression model was constructed to re-evaluate the effect of CD39 on HNSCC poor prognosis.Finally,Western blotting was used to detect the expression levels of CD39 protein in UM1 cell treated with dose-laddered dexamethasone(DXM),and analyze the relationship between the expression level of CD39 protein and the exposure dose of DXM in vitro.Results:The expression level of CD39 mRNA was obviously up-regulated in HNSCC tissues than in BM tissues(fold change: 1.618,P<0.01)from the analysis of Oncomine data.It was showed that CD39 protein was mainly expressed in tumor cells from the method of IHC,and 53(62.4%)of patients in 85 HNSCC were CD39 high expression.The expression level of CD39 protein was closely associated with preoperative chemotherapy(odds ratio [OR]=3.928,95% confidential interval [CI]: 1.676～9.206;P<0.01)and constant or heavy alcohol consumption(OR=0.294,95%CI: 0.111～0.775;P<0.05),while not significantly correlated with tumor size,lymph node metastasis,TNM pathological stage,location of primary tumor,mixed poor differentiated components,as well as age,gender,and smoking index of patients(all P>0.05).The high expression of CD39 in cancer tissues was an independent relapse risk factor(hazard ratio [HR]=2.328,95% CI=1.091～4.967;P<0.05)for patents with HNSCC.The expression level of CD39 protein in HNSCC tissues was positively correlated with the expression levels of PD-L1,PD-L2 and ALDH1A1 proteins(all P<0.05),while not significantly correlated with the distribution of CD3,CD8,CD103 and Foxp3 positive immunocytes(all P>0.05).The determinate factor(extracted from variables CD39,PD-L1,PD-L2 and ALDH1A1),T-staging and N-staging were the independent influence factors of RFS in patient with HNSCC(all HR>1,P<0.05).Additionally,CD39 protien was expressed in many a HNSCC cell,and DXM could enhance,in a dose-dependent manner,the expression of CD39 in UM1 cell in vitro.Conclusion:CD39 is over-expressed in HNSCC and is influenced by alcohol consumption and preoperative chemotherapy(possibly associated with DXM).The high expression of CD39 is an independent risk factor of recurrence for patients,suggesting that CD39 can be used as a potential bio-marker of patient with HSNCC.The expression level of CD39 is closely related to PD-L1,PD-L2 and ALDH1A1,suggesting that HNSCC may escape immune surveillance through many a mechanism.