Expression And Biological Significance Of Ezrin, CD44v6, E-cadherin And MMP-2 In Esophageal Carcinoma

Objective : To detect the expressions of Ezrin, CD44v6,E-cadherin,MMP-2 protein and Ezrin mRNA in Esophageal carcinoma, the relationship of Ezrin, CD44v6, E-cadherin and MMP-2 expression with the carcinogenesis, differentiation, invasion, metastasis of esophageal carcinoma is investigated; the relationship of Ezrin expression with CD44v6 , E-cadherin and MMP-2 protein expression in the development of esophageal carcinoma is investigated.Methods1 The expressions of Ezrin protein, CD44v6 protein, E-cadherin protein, and MMP-2 protein were examined by immunohistochemistry (IHC) in 45 cases of esophageal carcinoma, 21cases of atypical hyperplasia of esophageal epithelium and 24 cases of incisal margin normal esophageal mucosa tissues.2 The expression of Ezrin mRNA was examined by in situ hybridyzation (ISH) in 45 cases of esophageal carcinoma, 21 cases of atypical hyperplasia of esophageal epithelium and 24 cases of incisal margin normal esophageal mucosa tissues.3 The expression of Ezrin protein was examined by flow cytometry (FCM) in 27 cases of esophageal carcinoma, 10 cases of atypical hyperplasia of esophageal epithelium and 8 cases of incisal margin normal tissues.Results1 Expression of Ezrin in various lesions of esophageal epithelium1.1 Immunohistochemistry methodThe expressions of Ezrin protein in 45 cases of esophageal carcinoma,21 cases of atypical hyperplasia of esophageal epithelium and 24 cases of incisal margin normal tissues were detected by IHC, the results showed that the expression of Ezrin protein in esophageal carcinoma (73.33%) was higher than that in para-tumorous(61.90%) and in incisal margin normal tissues (8.33%), the difference was significant (P<0.05); the expression of Ezrin protein had no correlation with histological differentation degree,age or sex (P>0.05); but the expression of Ezrin protein had correlation with lymph node metastasis(P<0.05), the positive expression of Ezrin protein in cases with lymph node metastasis(85.19%) was higher than those without lymph node metastasis(55.56%). And the expression of Ezrin protein was correlated with the depth of invasion (p<0.05).1.2 In situ hybridyzation (ISH)The expressions of Ezrin mRNA in 45 cases of esophageal carcinoma, 21 cases of atypical hyperplasia of esophageal epithelium and 24 cases of incisal margin normal tissues were detected by ISH, the results showed that there was no the expression of Ezrin mRNA in incisal margin normal esophageal mucosa epithelium, the result was coincidental with the result of immunohistochemistry method. The expression of Ezrin mRNA in esophageal carcinoma was 71.11%,and the expression of Ezrin mRNA in esophageal para-tumorous was 42.89%; The expression of Ezrin mRNA was not significantly related with histological differentation degree,age or sex (p>0.05);but was correlated with lymph node metastasis and the depth of invasion (p < 0.05). The results were coincidental with the results of immunohistochemistry method.1.3 Flow cytometryThe expressions of Ezrin protein in 27 cases of esophageal carcinoma and 10 cases of atypical hyperplasia of esophageal epithelium and 8 cases of incisal margin normal tissues were detected by Flow cytometry, the results showed that the expression of Ezrin protein in esophageal carcinoma(1.5502±0.1055) and in esophageal para-tumors(1.1799±0.1781) were significantly higher than that in incisal margin normal tissues (1.008±0.2133) (p<0.05); there was a difference in cases with lymph node metastasis(0.36±0.12) and cases without lymph node metastasis(0.68±0.13)(p<0.05) and there was a difference in cases with the depth of invasion(p<0.05). But the expression of Ezrin protein wasn't related with the histological differentation degree, age or sex (p>0.05). All of the results were coincidental with the results of immunohistochemistry method.2 Expression of CD44v6 in various lesions of esophageal epitheliumThe expressions of CD44v6 protein in 45 cases of esophageal carcinoma,21 cases of atypical hyperplasia of esophageal epithelium and 24 cases of incisal margin normal tissues were detected by IHC, the results showed that the expression of CD44v6 protein in esophagealcarcinoma(66.67%)and in esophageal para-tumorous(42.86%) were higher than that in incisal margin normal tissues (0%), the difference was significant (P<0.05); the expression of CD44v6 protein had correlation with lymph node metastasis (P<0.05), the positive expression of CD44v6 protein in cases with lymph node metastasis(85.19%) was higher than those without lymph node metastasis(38.89%). But the expression of CD44v6 protein wasn't correlated with histological differentation degree , the depth of invasion ,age or sex(p>0.05).3 Expression of E-cadherin in various lesions of esophageal epitheliumThe expressions of E-cadherin protein in 45 cases of esophageal carcinoma,21 cases of atypical hyperplasia of esophageal epithelium and 24 cases of incisal margin normal tissues were detected by IHC, the results showed that the expression of E-cadherin protein in esophageal carcinoma (28.89%) and in esophageal para-tumorous(71.43%) were lower than that in incisal margin normal tissues (100%), the difference was significant (P<0.05); the expression of E-cadherin protein had positive correlation with histological differentation degree (P<0.05), the higher the differentation was, the higher the positive expression of E-cadherin protein was; and the expression of E-cadherin protein had correlation with lymph node metastasis(P<0.05), the positive expression of E-cadherin protein in cases with lymph nodemetastasis(7.41%) was lower than those without lymph node metastasis(61.11%). But the expression of E-cadherin protein wasn't correlated with the depth of invasion, age or sex (p>0.05).4 Expression of MMP-2 in various lesions of esophageal epitheliumThe expressions of MMP-2 protein in 45 cases of esophageal carcinoma,21 cases of atypical hyperplasia of esophageal epithelium and 24 cases of incisal margin normal tissues were detected by IHC, the results showed that the expression of MMP-2 protein in esophageal carcinoma(40.00%)and in esophageal para-tumorous(38.10%) were higher than that in incisal margin normal tissues (4.17%), the difference was significant (P<0.05); the expression of MMP-2 protein had positive correlation with histological differentation degree (P<0.05), the higher the differentation was, the higher the positive expression of MMP-2 protein was;the expression of MMP-2 protein had correlation with lymph node metastasis (P<0.05), the positive expression of MMP-2 protein in cases with lymph node metastasis(55.56%) was higher than those without lymph node metastasis(16.67%). And the expression of MMP-2 protein was correlated with the depth of invasion (p<0.05). But the expression of MMP-2 protein wasn't correlated with age or sex(p>0.05).5 The correlation analysis of Ezrin and CD44v6, Ezrin and E-cadherin, Ezrin and MMP-2 expressionsThere were positive correlation(X2=4.602 , P<0.05) between the expression of Ezrin and CD44v6 while negative correlation(X2=6.906,P<0.05) between the expression of Ezrin and E-cadherin in the esophageal carcinoma tissues. There were no significant correlation(X2=3.712,P>0.05) between the expression of Ezrin and MMP-2 in the esophageal carcinoma tissues.6 The correlation analysis of CD44v6 and E-cadherin, CD44v6 and MMP-2 expressionThere were negative correlation(X2=6.544,P < 0.05) between the expression of CD44v6 and E-cadherin while no significant correlation(X2=0.416 , P>0.05) between the expression of CD44v6 and MMP-2 in the esophageal carcinoma tissues.7 The correlation analysis of E-cadherin and MMP-2 expressionThere was negative correlation between the expression of E-cadherin and MMP-2 in the esophageal carcinoma tissues(X2=4.615,P<0.05).Conclusions1 The expressions of Ezrin and Ezrin mRNA in esophageal carcinoma tissues and atypical hyperplasia of esophageal epithelium were higher than that in incisal margin normal tissues and the expressions of Ezrin protein in cases with lymph node metastasis were higher than that without lymph node metastasis, which indicate that the high expression of Ezrin could involve in esophageal epithelium carcinogenesis and metastasis of esophageal carcinoma.2 The expressions of CD44v6 protein in esophageal carcinoma tissues and atypical hyperplasia of esophageal epithelium were higher than that in incisal margin normal tissues and the expressions of CD44v6 protein in cases with lymph node metastasis were higher than that without lymph node metastasis, which indicate that the high expression of CD44v6 protein could involve in esophageal carcinogenesis and metastasis of esophageal carcinoma.3 The expressions of E-cadherin protein in esophageal carcinoma tissues and atypical hyperplasia of esophageal epithelium were lower than that in incisal margin normal tissues and the expressions of E-cadherin protein in cases with lymph node metastasis were lower than that without lymph node metastasis, which indicate that the low expression of E-cadherin could involve in esophageal carcinogenesis and metastasis of esophageal carcinoma; the expressions of E-cadherin protein had positive correlation with histological differentation, it indicates that there is some reference value of expression of E-cadherin protein to the degree of esophageal carcinoma's malignancy.4 The expressions of MMP-2 protein in esophageal carcinoma tissues and atypical hyperplasia of esophageal epithelium were higher than that in incisal margin normal tissues and the expressions of MMP-2 protein in cases with lymph node metastasis were higher than that without lymph node metastasis, which indicate that the high expression of MMP-2 protein could involve in esophageal carcinogenesis and metastasis of esophageal carcinoma; the expressions of MMP-2 protein had positive correlation with histological differentation, it indicates that there is some reference value of expression of MMP-2 protein to the degree of esophageal carcinoma's malignancy.5 There were positive correlation between the expression of Ezrin and CD44v6 while negative correlation between the expression of Ezrin and E-cadherin in the esophageal carcinoma tissues, which indicate that there may be a cooperation between Ezrin and CD44v6 in the progression of esophageal carcinoma while a inhibition between Ezrin and E-cadherin in the suppression of esophageal carcinoma metastasis6 There were negative correlation between the expression of E-cadherin and CD44v6, E-cadherin and MMP-2 in the esophageal carcinoma tissues, which indicate that there may be inhibitions between E-cadherin and CD44v6, E-cadherin and MMP-2 in the suppression of esophageal carcinoma metastasis.