Expression And Clinical Significance Of CXCL13 And CXCR5 In Hypopharyngeal Squamous Cell Carcinoma

Objective Although hypopharyngeal squamous cell carcinoma is rare in head and neck tumors,and the incidence is relatively low,its malignant degree is high,focusing on survival rate and organ function(laryngeal and pharynx function)is the focus of hypopharyngeal squamous cell carcinoma treatment.In the treatment of patients with HSCC,the most worthy of further research is how to avoid damaging laryngeal function while ensuring the survival rate and improving the quality of survival of patients.Although the treatment methods for patients with hypopharyngeal squamous cell carcinoma are continuously improved and developed,so far,the survival rate of hypopharyngeal cancer is still difficult to be significantly improved.As a small molecule protein polypeptide,the chemokine CXCL13 acts in combination with the CXCR5 receptor of its immune receptor,which can directly promote angiogenesis,tumor cell growth,survival and differentiation and tumor spread,and can affect the early generation and development process of tumor cells.The purpose of this study is to explore the expression and clinical significance of CXCL13 and CXCR5 in hypopharyngeal squamous cell carcinoma,and hope to contribute to the treatment of hypopharyngeal squamous cell carcinoma in the future.Methods The specimens of pathological changes and adjacent normal tissues of 25 patients with hypopharyngeal squamous cell carcinoma diagnosed by pathology after the first operation in the Affiliated Hospital of Qingdao University were collected.The in vitro expression level and clinical significance of CXCL13 and CXCR5 genes in hypopharyngeal squamous cell carcinoma were studied by real-time fluorescence quantitative PCR and Western blot.1.Quantitative real-time PCR(RT-q PCR):Samples of hypopharyngeal cancer tissues and adjacent normal tissues from 25 patients were used for RT q PCR experiments.Differences in expression of CXCL13 m RNA and CXCR5 m RNA were investigated by paired samples t-test,and associations between CXCL13 m RNA and CXCR5 m RNA and hypopharyngeal squamous cell carcinoma patients were investigated by one-way ANOVA and independent samples t-test.The correlation of CXCL13 and CXCR5 was analyzed by Pearson correlation analysis.2.Protein blotting(Western Blot,WB): The bands obtained from 25 patients with hypopharyngeal carcinoma lesions and normal adjacent tissues samples by WB technology were measured by Image J,and the association of CXCL13 protein,CXCR5 protein and hypopharyngeal squamous cell carcinoma were explored using the Mann-Whitney U rank-sum test or Kruskal-Wallis test.Spearman rank correlation was used to analyze the relationship between the expression of CXCL13 protein and CXCR5 protein.Results 1.RT-q PCR results: the relative expression of CXCL13 m RNA and CXCR5 m RNA was significantly higher in hypopharyngeal squamous cell carcinoma tissues than in adjacent normal tissues.Relative expression of CXCL13 m RNA t=-12.415,P=0.000,and CXCR5 m RNA t=-11.759 and P=0.000.The relative expression of CXCL13 m RNA and CXCR5 m RNA was significantly different in clinical stages(early metaphase and late stage),whether lymph node metastasis,degree of pathological differentiation(P<0.05),but no expression difference in age and primary site(P> 0.05).The Pearson correlation analysis showed a significant positive correlation between the relative expression of CXCL13 m RNA and CXCR5 m RNA,at r=0.853 and P=0.000 in hypopharyngeal squamous cell carcinoma patients.2.Western Blot results: The relative expression of CXCL13 and CXCR5 protein was significantly higher in hypopharyngeal squamous cell carcinoma tissues than in adjacent normal tissues,with Z=-6.065,P=0.000,Z=-5.910 and P=0.000.Moreover,the relative expression of CXCL13 and CXCR5 protein was consistent with the m RNA in each clinical and pathological feature group.Spearman correlation analysis showed the relative expression of CXCL13 and CXCR5 protein r=0.818 and P=0.000 in hypopharyngeal squamous cell carcinoma patients.Conclusion 1.The m RNA and protein levels of CXCL13 and CXCR5 were significantly higher in hypopharyngeal squamous cell carcinoma tissues compared with adjacent normal tissues,indicating that both CXCL13 and CXCR5 may promote tumor progression in hypopharyngeal squamous celcarcinoma.2.Expression of CXCL13 and CXCR5 m RNA and protein levels was highly expressed in poorly differentiated,late clinical and groups with lymph node metastasis,suggesting that the expression of CXCL13 and CXCR5 was associated with the development,metastasis and malignancy of hypoopharyngeal carcinoma.3.The m RNA levels and protein levels of CXCL13 and CXCR5 genes can be highly expressed in hypopharyngeal cancer cells and show a significant positive correlation with them.The research on both of them can clarify the potential benefits of targeting CXCL13/CXCR5 pathway,and thus play a guiding role in the treatment of hypopharyngeal cancer.