| Research background:Laryngeal cancer is one of the most common malignant tumors of the head and neck,and in recent years,the prevalence of laryngeal cancer is increasing.Histopathological types of the laryngeal squamous cell carcinoma is diverse,and the squamous cell carcinoma is the most common,which accounting for about 95%.The treatment for people who have disese is various,for example: surgery,radiotherapy and chemotherapy,but the prognosis are not ideal.Local recurrence,cervical lymph node metastasis and poor differentiation are the main reasons for the bad treatment of the patients and the poor prognosis.Therefore,the focus of the study is the development characteristics and differences of laryngeal squamous cell carcinoma in different an homhistological types and metastatic lesions.Enhancer of Zeste Homolog 2(EZH2),as a humolog of zeste genetic enhancer in fruit flies and belonging to the poly-comb(PcG)group,methylates the 27 th lysine of H3 in nucleosome histone(H3K27).Methylation modification of histone H3K27(H3K27me3)always inhibits transcription.At present,many studies have found that EZH2 expression is increased in epithelial cell carcinoma.It is reported that high expression of EZH2 is closely related to tumor growth,lymph node metastasis,clinical stage and poor prognosis.In recent study,EZH2 is highly expressed in breast cancer,prostate cancer,bladder cancer and liver cancer and other malignant tumors,and its high expression is closely related to tumorigenesis,cell proliferation,invasion,metastasis and prognosis of patients.However,the expression of EZH2,H3K27me3 and E-cadherin in laryngeal squamous cell carcinoma is relatively rare,so we want to detect the expression of EZH2,H3K27me3 and E-cadherin in laryngeal carcinoma by immunohistochemistry and real-time fluorescence quantitative PCR respectively.The purpose of this study was to explore the possible role of its occurrence,development,metastasis and different histological types in laryngeal squamous cell carcinoma,to provide new targets for the diagnosis,treatment and prognosis of laryngeal squamous cell carcinoma and to improve the therapeutic effect of the patients and prognosis.Objective:To investigate the expression of EZH2,H3K27me3 and E-cadherin in laryngeal squamous cell carcinoma,to explor the relashionship of EZH2,H3K27me3 and E-cadherin among age,histological differentiation,clinical stage,clinical classification and lymph node involvement,respectively.Then analyze their role in the tumorigenesis,progession,invasion and metastasis as well as their correlation.Methods:1.Immunohistochemistry was employed to detected the expressions of EZH2,H3K27me3 and E-cadherin in 45 cases of laryngeal squamous cell carcinoma and corresponding 45 cases of adjacent tissues.2.qRT-PCR was employed to detected the expression of EZH2 and E-cadherin in 24 cases of laryngeal squamous cell carcinoma and corresponding adjacent tissues.3.Statistics: SPSS19.0 was applied to analyze the results of experiment.P <0.05 indicating significant difference.Results:1 Expression of EZH2 and relation with clinical parameter1.1: Immunohistochemistry was employed to detected the expressions of EZH2 in 45 cases of laryngeal squamous cell carcinoma and corresponding 45 cases of adjacent tissues.The mean optical density values of EZH2 in LSCC were 0.383 ± 0.129,while the mean optical density values of adjacent tissues were 0.208 ± 0.038.There was significant difference between the two groups.(P = 0.000).EZH2 protein expression in laryngeal squamous cell carcinoma was significantly higher than adjacent tissues.Expression of EZH2 was correlated with lymphnode metastasis(N1>N0,P = 0.000),clinical stages(stage ??? >stage ???,P = 0.001),pathological grading of tumor(poorly differentiated expression was higher than well differentiated,P = 0.008),but there was not correlated with clinical classification(P = 0.466)and age(P = 0.793).1.2: qRT-PCR was employed to detected the expression of EZH2 mRNA in 24 cases of laryngeal squamous cell carcinoma and corresponding adjacent tissues,and the expression of EZH2 mRNA were(1.533 ± 3.389)and(0.591 ± 1.641).There is statistic significance between them,and the formor was higher than the latter(P = 0.001).The expression of EZH2 mRNA was correlated with lymph node metastasis(N1>N0,P = 0.045),and tumor pathological grade(lower middle differentiation higher than higher differentiation,P = 0.029),but there was not correlated with clinical classification(P = 0.297),clinical stage(P = 0.278)and age(P = 0.896).2 H3K27me3 expression and its relationship with clinical parameters2.1: Immunohistochemistry was employed to detected the expressions of H3K27me3 in 45 cases of laryngeal squamous cell carcinoma and corresponding 45 cases of adjacent tissues.The mean optical density values of H3K27me3 in LSCC were 0.380 ± 0.090,while the mean optical density values of adjacent tissues were 0.173 ± 0.154.There was significant difference between the two groups.(P = 0.000).H3K27me3 protein expression in laryngeal squamous cell carcinoma was significantly higher than adjacent tissues.Expression of H3K27me3 was correlated with lymphnode metastasis(N1>N0,P = 0.000),clinical stages(stage ? and ? >stage ? and ?,P = 0.000),and pathological grading of tumor(poorly differentiated expression was higher than well differentiated,P = 0.009),but there was not correlated with clinical classification(P = 0.897)and age(P = 0.892).3 E-cadherin expression and its relationship with clinical parameters3.1: Immunohistochemistry was employed to detected the expressions of E-cadherin in 45 cases of laryngeal squamous cell carcinoma and corresponding 45 cases of adjacent tissues.The mean optical density values of E-cadherin in LSCC were 0.341 ± 0.114,while the mean optical density values of adjacent tissues were 0.399 ± 0.062.There was significant difference between the two groups.(P = 0.007).E-cadherin protein expression in laryngeal squamous cell carcinoma was significantly lower than adjacent tissues.Expression of E-cadherin was correlated with lymphnode metastasis(N0>N1,P = 0.013),clinical stages(stage?and?were higher than stage ? and ?,P = 0.001),and pathological grading of tumor(well differentiated expression was higher than poorly differentiated,P = 0.000),but there was not correlated with clinical classification(P = 0.87)and age(P = 0.108).3.2: qRT-PCR was employed to detected the expression of E-cadherin m RNA in 24 cases of laryngeal squamous cell carcinoma and corresponding adjacent tissues.,the expression of E-cadherin mRNA were(1.837 ± 1.663)and(5.284 ± 6.789),there is statistic significance between them,the formor was lower than the latter(P = 0.003).And the expression of E-cadherin mRNA was correlated with lymph node metastasis(N0>N1,P = 0.046),and tumor pathological grade(well differentiation higher than poorly differentiation,P = 0.027),but there was not correlated with clinical classification(P = 0.297),clinical stage(P = 0.886)and age(P = 0.403).4 There correlations in protein and mRNA4.1 At the protein level,both EZH2 and E-cadherin?H3K27me3 and E-cadherin expression were negatively correlated;while EZH2 and H3K27me3 expression was positively correlated.4.2 At the genetic level,EZH2 and E-cadherin m RNA expression was negatively correlated.Conclusion:1.EZH2,H3K27me3 and E-cadherin may play important roles in the occurrence,development,invasion and metastasis of laryngeal squamous cell carcinoma.2.The expressions of EZH2,H3K27me3 and E-cadherin were closely related to the clinical stage,histopathological grade and lymph node involvement in laryngeal squamous cell carcinoma.3.Combined detection of EZH2,H3K27me3 and E-cadherin expression in laryngeal squamous cell carcinoma is of great significance in predicting the occurrence and development of laryngeal squamous cell carcinoma as well as invasion and metastasis. |
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