Expression And Significance Of Integrinβ3,MMP-2 And VEGF In Esophageal Squamous Cell Carcinoma

Background and objectiveEsophageal carcinoma (EC) is one of the sixth most common malignant tumoursin the world. Multiple factors and pathways cause the carcinogenesis and development of the esophageal carcinoma. In recent years, with the development of molecular biology, the pathogenesis of the esophageal carcinoma has been known deeply, but its true molecular mechanism is not totally figured out.In this study , the expressions of integrinβ3 protein MMP-2 protein and VEGF protein in esophageal squamous cell carcinoma , dysplasia and corresponding normal epithelium tissues were detected by immunohistochemistry. The relationship between them and the carcinogenesis, invasion and lymph node metastasis of esophageal squamous cell carcinoma were explored. This aim was to investigate the function of integrinβ3 protein MMP-2 protein and VEGF protein and the correlations of them in carcinogensis, development, invasion and metastasis of esophageal squamous cell carcinoma, in order to find out useful ways to suppressoccurrence and development of esophageal squamous cell carcinoma.Materials and methods1. Sample: 41 cases of esophageal carcinoma were collected from the FirstAffiliated Hospital, Zhengzhou University in 2002. All cases had no the history of the chemotherapy, radiotherapy and immunotherapy. Two pathologists identified all cases, which was entirely esophageal squamous cell carcinoma. 22 cases were I rank, 13 cases were II rank, 6 cases were III rank of histological grade. 12 cases were in the low layer invaded and 29 cases in the deep layer invaded. 22 examples appeared the lymph node metastasis, 19 cases were non-lymph node metastasis. Each of all in site of tumor and tumor-adjacent mucosa cut pieces of tissue involved of simple proliferation in 17 cases, mild dysplasia in 13 cases, severe dysplasia in 10 cases, carcinoma in situ in 6 cases and normal esophageal epithelium in 19 cases.2. Reagent: Polyclonal anti-integrinβ3 antibody was purchased from Boster Biotech Company, Polyclonal anti-MMP-2 antibody and monoclonal antibody against VEGF were both purchased from Zhonghshan Biotech Company, Beijing.The SP staining kit was bought from Zhonghshan Biotech Company, Beijing.3. Method: Streptomycete avidin peroxides (SP) immunohistochemistry was used to examine the expression of integrinβ3 protein, MMP-2 protein and VEGF protein in tissues of 41 cases of esophageal squamous cell carcinoma and precancerouslesions.Results1. The positive rate of integrinβ3 protein in esophageal squamous cell carcinoma(82.9%, 34/41) was significantly higher than those in normal mucosa (21.1%, 4/19) (P< 0.05) . With the increase of invasion depth , the positive rate of expression of integrinβ3 protein in esophageal squamous cell carcinoma was increased, the positive rates of integrinβ3 protein in low layer invaded group and deep invaded group of esophageal squamous cell carcinoma were 50.0%( 6/12 )and 96.6%( 28/29), respectively. There was a significant difference between them (P<0.05). The positive rates of integrinβ3 protein in the lymph node metastasis group and nonlymph node metastasis group were 95.5% (21/22) and 68.4% (13/19) , respectively. There was a significant difference between them(P<0.05), too. The positive rates of integrinβ3 protein in tumor-adjacent normal epithelium, simple hyperplasia epithelium, mild atypical hyperplasia epithelium, severe atypical hyperplasia epithelium , carcinoma in situ were increased gradually (21.1% 64.7%, 69.2% 80.0%, 100.0%, respectively), with significant difference between the simple hyperplasia epithelium, mild atypical hyperplasia epithelium, severe atypical hyperplasia epithelium, carcinoma in situ and the normal epithelium. While there was no significant difference in the positive rate of integrinβ3 protein in different differentiation of carcinoma.2. The positive rate of MMP-2 protein in esophageal squamous cell carcinoma (68.3%, 28/41) was significantly higher than that in adjacent normal mucosa (5.3%, 1/19) (P< 0.05) . The positive rate of MMP-2 protein in mild atypical hyperplasia epithelium, severe atypical hyperplasia epithelium and carcinoma in situ were 41.7% 80.0% 100.0%, respectively, with significant difference between the mild atypical hyperplasia epithelium, severe atypical hyperplasia epithelium, carcinoma in situ and the normal epithelium. The positive rates of MMP-2 protein in low layer invaded group and deep layer invaded of esophageal squamous cell carcinoma group were 41.7% (5/12) and 79.3% (23/29) , respectively. There was a significant difference between them (P<0.05). The positive rates of MMP-2 protein in the lymph node metastasis group and nonlymph node metastasis group were 81.8% (18/22) and 52.6%(10/19) , respectively. There was a significant difference between them (P<0.05), too. While there was no significance difference in the expression of MMP-2 protein in different differentiation of carcinoma, although the positive rate was higher gradually (P>0.05).3. The positive rate of VEGF protein in esophageal squamous cell carcinoma (48.8%, 20/41) was significantly higher than that in adjacent normal mucosa (0.0%, 0/19) (P< 0.05) . The positive rates of VEGF protein in tumor-adjacent normal epithelium, simple hyperplasia epithelium, mild atypical hyperplasia epithelium, severe atypical hyperplasia epithelium, carcinoma in situ were increased gradually( 0.0% 23.5% 69.2% 80.0% 83.3%, respectively). There was a significant difference between the mild atypical hyperplasia epithelium, severe atypical hyperplasia epithelium, carcinoma in situ and the normal epithelium (P< 0.05). The positive rates of VEGF protein in low layer invaded group and deep layer invaded of esophageal squamous cell carcinoma group were 25.0% (3/12) and 58.6% (17/29) , respectively, with significant difference between them (P<0.05). The positive rates of VEGF protein in the lymph node metastasis group and nonlymph node metastasis group were 63.6% (14/22) and 31.6% (6/19), respectively. There was a significant difference between them(P<0.05), too. There was no significant difference in different differentiation grades of carcinoma tissues (P> 0.05) , although the positive rate was higher gradually.4. There was a positive correlation between the positive expression rate of integrinβ3 protein and that of MMP-2 protein in esophageal squamous cell carcinoma. There was 27 cases positive expression of MMP-2 protein in 34 cases positive expression of integrinβ3 protein, there was 6 cases negative expression of MMP-2 protein in 7 cases negative expression of integrinβ3 protein.5. The positive expression of integrinβ3 protein was positively correlated with that of VEGF protein in esophageal squamous cell carcinoma, there was 17 cases positive expression of VEGF protein in 34 cases positive expression of integrinβ3 protein, there was 4 cases negative expression of VEGF protein in 7 cases negative expression of integrinβ3 protein.6. There was a positive correlation between the positive expression rate of MMP-2 protein and VEGF protein in esophageal squamous cell carcinoma, there was 15 cases positive expression of VEGF protein in 28 cases positive expression of MMP-2 protein, there was 8 cases negative expression of VEGF protein in 13 cases negativeexpression of MMP-2 protein.Conclusion1. The positive expression of integrinβ3 protein in tumor-adjacent atypicalhyperplasia epithilum had a trend of increase gradually, which showed that over expression of integrinβ3 protein might play an important role in the carcinogenesis of esophageal squamous cell carcinoma. The expression of integrinβ3 protein was related to invasion and lymph node metastasis, it was indicated that integrinβ3 protein was correlated with invasion and lymph node metastasis of esophageal squamous cell carcinoma.2. The overexpression of MMP-2 protein in carcinoma and tumor-adjacent tissues suggested that it maybe play an important role in the carcinogenesis and development of esophageal squamous cell carcinoma.3. The overexpression of VEGF protein was correlated with the occurrence, invasion and metastasis of esophageal squamous cell carcinoma.4. There was a positive correlation among the expression of integrinβ3 protein, MMP-2 protein and VEGF protein, which implied that the positive expression of integrinβ3 protein could ascending the expression of MMP-2 protein and VEGF protein.5. United detection of integrinβ3 protein, MMP-2 protein and VEGF protein could become an objective target to evaluate the invasion and metastasis of esophageal squamous cell carcinoma. It had a great significance to judge the prognosis of esophageal squamous cell carcinoma.