Expression And Significance Of NF-κB P50, P65, MMP-9 And ICAM-1 In Esophageal Squamous Cell Carcinoma

Background and objectiveEsophageal carcinoma is a kind of common maglignant tumor in china. It was the fourth in the death rate of malignant tumor in our country. The most common reasons leading to the death of esophageal carcinoma patients are the tumor's invasion and metastasis. So explorating the mechanism about invasion and metastasis of esophageal carcinoma has become a focus.Nuclear transcription factor-κB (NF-κB) is an important type of nuclear transcription factor. The main form of NF-κB is a dimmer composed of p50 and p65 protein, the former is the part where NF-κB is linked with DNA, and the latter takes part in the primary regulation of gene's transcription and enhances the linkage between p50 and DNA. It can regulate the expression of many genes and plays an important role in the carcinogenesis and development of the tumor. Matrix matalloproteinases (MMPs) belongs to endopeptidase family, which contains zinc ionic. It can degrade the extrecelluer matrix and play important roles in the infiltration and metastasis of the malignant tumor. Matrix metalloproteinases-9 (MMP-9) belongs to gelatinase. Intercellular adhesion molecule l(ICAM-l) can regulate the adhension between cells or the cell and extracellular matrix and play important roles in the infiltration and metastasis of the tumor.It has been reported that in many tumors, such as mammary adenocarcinoma, gastric carcinoma, lung cancer and so on, the activation of NF-κB can up-regulate the expression of MMP-9 and ICAM-1 protein and promote the infiltration and metastasis of the tumor. But there was no report about the relations between NF-κB and MMP-9 or ICAM-1 in esophageal squamous cell carcinoma. This study will detect the expression of NF-κB p50, p65 protein and mRNA in specimens of esophageal squamous cell carcinoma and tumor-adjacent mucosas (inclouding carcinoma in situ, atypical hyperplasia epithelium and normal epithelium) by the technique of immunohistochemistry and in situ hybridization, and the expression of MMP-9 and ICAM-1 protein in specimens of esophageal squamous cell carcinoma and tumor-adjacent mucosas by immunohistochemistry. And we will explore the relationships among them.Materials and methods49 cases of esophageal squamous cell carcinoma were collected in the First Affiliated Hospital, Zhengzhou University. All the cases had no history of the chemotherapy, radiotherapy and immunotherapy. Pathologist had identified all the specimens. All of them are entirely esophageal squamous cell carcinoma. 24 cases are grade I, 16 cases are grade II, 9 cases are grade III. 23 cases had the lymph node metastasis, 26 cases non-lymph node metastasis. 13 cases are on the low layer invaded and 36 cases are on the deep layer invaded. Each was cut in the site of tumor and the tumor-adjencent mucoma. The expressions of NF-κB p50, p65 mRNA and NF-κB P50, P65, MMP-9 and ICAM-1 protein were detected in 49 cases of esophageal squamous cell carcinoma, and tumor-adjacent mucosa tissues by in situ hybridization and immunohistochemistry. All the datas were analyzed by SPSS10.0 stastistical package; the count information calculated the positive rate. The comparision of positive rate uses the Chi-square; the mean of more than two groups uses ANVOA. The relation of two variables is analyzed by Spearman's correlation analysis. The level of significant difference was a=0.05.Results 1. The positive rates of expression of NF-κB p50 protein in tumor-adjacent normal epithelium and atypical hyperplasia epithelium, carcinoma in situ and invasive carcinoma were 28.1%, 36.8%, 33.3%, 55.1% respectively. There was a significant difference between invasive carcinoma and the normal epithelium (P<0.05). The positive rates of the expression of NF-κB p65 protein in tumor-adjacent normal epithelium and atypical hyperplasia epithelium, carcinoma in situ and invasive carcinoma were 18.8%, 42.1%, 50.0%, 57.1% respectively. And there was a significant difference between invasive carcinoma and the normal epithelium (P<0.05).2. The positive rates of expression of p50 protein in grade I, II and III of esophageal squamous cell carcinoma were 37.5%, 62.5%, 88.9% respectively, a significant difference among them(P<0.05). In the tissues of esophageal squamous cell carcinoma, the positive rates of expression of NF-κB p50 protein on the low layer invaded and the deep layer invaded were 15.4% and 69.4% respectively, a significant difference between them (P<0.05). The positive rate of expression of NF-κB p50 protein in the group with lymph node metastasis was 60.9%. While in the group without lymph node metastasis, it was 50.0%, no significant difference between them (P>0.05). The positive rates of expression of NF-κB p65 protein in esophageal squamous cell carcinoma of grade I, II and III were 41.7%, 68.8% and 77.8% respectively, no significant difference among them (P>0.05). In the tissues of esophageal squamous cell carcinoma, the positive rates of expression of NF-κB p65 protein on the low layer the invaded and deep layer invaded were 7.7% and 75.0% respectively, a significant difference between them (P<0.05). The positive rate of expression of NF-κB p65 protein in the group with lymph node metastasis was 87.0%. While in the group without lymph node metastasis, it was 30.8%, a significant difference between them (P<0.05).3. The positive rates of the expression of NF-κB p50 mRNA in tumor-adjacent normal epithelium, atypical hyperplasia epithelium, carcinoma in situ and invasive carcinoma were 21.9%, 31.6%, 41.7%, 53.1% respectively. There was a significant difference between invasive carcinoma and the tumor-adjacent normal epithelium (P<0.05). The nuclear positive rates of the expression of NF-κB p65 mRNA in tumor-adjacent normal epithelium and atypical hyperplasia epithelium, carcinoma in situ and invasive carcinoma were 18.8%, 36.8%, 33.3%, 55.1% respectively. And there was a significant difference between invasive carcinoma and the tumor-adjacent normal epithelium (P<0.05).4. The positive rates of the expression of p50 mRNA in grade I,II and III of esophageal squamous cell carcinoma were 37.5%, 62.5%, 77.8% respectively, no significant difference among them (P>0.05). In the tissues of esophageal squamous cell carcinoma, the positive rates of expression of NF-κB p50 mRNA on the low layer invaded and the deep layer invaded were 15.4%, 66.7%) respectively, a significant difference between them (P<0.05). The positive rate of the expression of NF-κB p50 mRNA in the group with lymph node metastasis was 78.3%. While in the group without lymph node metastasis, it was 30.8%. There was a significant difference between them, too (P<0.05). The positive rates of expression of NF-κB p65 mRNA in esophageal squamous cell carcinoma of grade I, II and III were 45.8%, 50.0%, 88.9% respectively, no significant difference among them (P>0.05). In the tissues of esophageal squamous cell carcinoma, the nuclear positive rates of expression of NF-κB p65 mRNA on the low layer the invaded and deep layer invaded were 69.4%, 15.4% respectively, a significant difference between them (P<0.05). The positive rate of expression of NF-κB p65 mRNA in the group with lymph node metastasis was 73.9%. While in the group without lymph node metastasis, it was 38.0%, a significant difference between them (P<0.05).5. The positive rates of the expression of MMP-9 protein in normal epithelium, atypical hyperplasia epithelium, carcinoma in situ and invasive carcinoma were 34.4%, 52.6%, 58.3%, 71.4% respectively, a significant difference between invasive carcinoma and the normal epithelium (P<0.05). The positive rates of expression of ICAM-1 protein in normal epithelium, atypical hyperplasia epithelium, carcinoma in situ and invasive carcinoma were 0.0%, 0.0%, 0.0%, 42.9% respectively, a significant difference between invasive carcinoma and tumor-adjacent mucosa (P<0.05).6. The positive rates of the expression of MMP-9 protein in the lymph node metastasis group and non-lymphnode metastasis group were 95.7% and 50.0% respectively, a significant difference between them (P<0.05). While the positive rates of the expression of MMP-9 protein in grade I, II and III of esophageal squamous cell carcinoma were 58.3%, 75.0%, 100.0% respectively, no significant difference among them (P>0.05). In the tissues of esophageal squamous cell carcinoma, the positive rates of the expression of MMP-9 protein on low layer invaded group and deep layer group invaded were 46.2% and 80.6% respectively, a significant difference between them (P<0.05). The positive rates of the expression of ICAM-1 protein in the lymph node metastasis group and nonlymph node metastasis group were 69.6% and 19.2% respectively, a significant difference between them (P<0.05). While the positive rates of the expression of ICAM-1 protein in grade I, II and III of esophageal squamous cell carcinoma were 16.7%, 68.8%, 66.7% respectively, a significant difference among them(P<0.05). In the tissues of esophageal squamous cell carcinoma, the positive rates of the expression of ICAM-1 protein on low layer invaded group and deep layer invaded group were 15.4% and 52.8% respectively, a significant difference between them (P<0.05).7. There were positive correlations between the positive expression rates of NF-κB p50, p65 mRNA and the expression rates of NF-κB p50, p65 protein in esophageal squamous cell carcinoma (P<0.05).8. There were positive correlations between the positive expression rates of NF-κB p50, p65 protein and the expression rates of MMP-9, ICAM-1 protein in esophageal squamous cell carcinoma (P<0.05).Conclusions1. By detecting the expression of NF-κB p50, p65 protein and mRNA in esophageal squamous cell carcinoma and tumor-adjacent tissues, we found that both the rates of the expression of NF-κB p50, p65 protein and mRNA in esophageal squamous cell carcinoma are higher than those in tumor-adjacent tissues. From normal epithelium, atypical hyperplasia epithelium, carcinoma in situ to invasive carcinoma, the positive rates of NF-κB p50, p65 protein expression show an increasing trend, which indicate that the expression of NF-κB may be correlated with the carcinogenesis of esophageal squamous cell carcinoma.2. There were positive correlations between the expression of NF-κB p65 protein and mRNA and the invasive depth, the lymph node metastasis of esophageal squamous cell carcinoma, which indicate that the activation of p65 may promote the invasion and metastasis of esophageal squamous cell carcinoma.3. The expression of MMP-9, ICAM-1 protein was associated with invasion depth and lymph node metastasis of esophageal squamous cell carcinoma, which implied that MMP-9 and ICAM-1 protein could reflect the potential power of invasion and lymph node metastasis of esophageal squamous cell carcinoma.4. The expression of NF-κB p50, p65 protein has positive correlations with the expression of MMP-9 and ICAM-1 protein, which showed that the activation of p50 and p65 could induce the high-speed growth, invasion and metastasis of the tumor through regulating the expression of MMP-9 and ICAM-1.