| According to the statistic data of American Cancer Society in2012, lung cancer has become the most common carcinoma and is the first cause of death. The proportion of non-small cell lung cancer (NSCLC) in lung cancer is approximately80%. Despite the advancement in surgical, radio-and chemo-therapies over the years, the5-year survival rate of lung cancer is still not significantly improved, and the majority of NSCLC patients are usually diagnosed during the later stages of lung cancer and die without timely treatment. Therefore, early detection and development of novel therapeutic approaches for the successful treatment of lung cancer are critical. Annexin A1(ANXA1), which involves in a variety of pathological and physiological process, such as the inflammatory response, cell differentiation and proliferation, cell death signal regulation and clearance of apoptotic cells, is a Ca2+-dependent phospholipid binding protein. In recent years, the researchers found that ANXA1plays an important role in the occurrence and development of many cancers like head and neck squamous cell carcinoma, esophageal squamous cell carcinoma, prostate cancer, liver cancer, colorectal cancer, and is associated with the malignant degree of tumor, including local invasion, distal metastasis and prognosis deterioration. However, the study of ANXA1expression in NSCLC is few. The Wnt/p-catenin signal pathway participates in many physiological and pathologic processes such as biological development, cellular transport and apoptosis. And in recent years, its important role in a variety of tumors become a hot spot in tumor research. In Wnt/β-catenin pathway, β-catenin, as a most critical factor, can regulate the Wnt pathway by changing its expression or its distribution in cells. Based on the previous work of our department, in this study, we detected the expression of ANXA1and β-catenin in NSCLC, expecting to provide the theoretical basis for the early diagnosis and looking for new treatment methods for NSCLC.Objective:This project intends1) to detect the expression of ANXA1and P-catenin in tumor tissue specimens and their distal tissues (from the edge of the tumor>5cm),2) to analyse the relationship between ANXA1expression and tumor development of NSCLC,3) to explore correlation of anxal and Wnt/β-catenin pathway in NSCLC development.Methods:64cases of pathologically diagnosed cancer tissues and their paried distal tissue were selected as experimental and control group, respectively. ANXA1and p-catenin mRNA were detected by using real-time quantitative polymerase chain reaction(RT-qPCR). ANXA1and β-catenin proteins were tested by using Western blotting and immunohistochemical staining. Datas were analyzed by SPSS17.0software packet, measurement data were compared with t-test or one-way ANOVA analysis, qualitative variables were compared with chi-square test, and correlation analysis were performed with Spearman’s rank correlation analysis. Statistical significance level was defined as P value<0.05.Results:1.Real-time PCR:①The mRNA expression of ANXA1in lung cancer tissues was higher than that of the distal cancerous tissues (0.574±1.403vs.0.240±0.893, t=2.060P=0.045). The difference of expression was closely related to degree of differentiation, lymph node metastasis and TNM staging (F=9.954P=0.000, t=2.186P=0.036, F=4.564P=0.015), but had no correlation with the. gender, age, smoking history, tumor diameter and histological type (P>0.05).②The mRNA expression of β-catenin in lung cancer tissues was higher than that of the distal cancerous tissues (0.059±0.130vs.0.033±0.078,t=2.125P=0.040). The difference of expression was closely related to degree of differentiation and lymph node metastasis (F =4.888P=0.014, t=-2.465P=0.023), but had no correlation with the gender, age, smoking history, tumor diameter, histological type and TNM stage (P>0.05).③At the mRNA level, there was a positive correlation between the expression of ANXA1and β-catenin in lung cancer tissues (r=0.368P=0.008).2.Immunohistochemical staining on paraffin sections showed that:①ANXA1protein was significantly higher expression in tumor cells than normal cells. In lung cancer tissues, ANXA1distributed not only in the cytoplasm and membrane, but also express in the macrophages of the tissue adjacent to carcinoma. The positive expression rate of ANXA1protein in lung cancer tissues was significantly higher than that in paracancerous tissues (76.6%(49/64) vs.3.1%(2/64)). The difference of expression was closely related with degree of differentiation, lymph node metastasis and TNM staging (/2=9.661P=0.008, χ2=6.050P=0.014,χ2=7.018P=0.030), but had no relation with the gender, age, smoking history, tumor diameter, histological type (P>0.05).②In normal tissues, a small amount of P-catenin protein expressed in the cytoplasm and membrane. However, there was a tendency that β-catenin was highly expressed in lung tissue, mainly expressed in the cell membrane, abnormal deposition in the nucleus could also be observed. The positive expression rate of P-catenin protein in lung cancer was higher than the distal tissue (71.9%(46/64) vs.3.1%(2/64),χ2=64.533P=0.000). The expression difference was closely related to gender, differentiation degree and lymph node metastasis (χ2=4.753P=0.029,χ2=11.077P=0.004,χ2=5.154P=0.023), but had no related with age, smoking history, tumor diameter, histological type and TNM stage (P>0.05).③At the protein level, there was a positive correlation between the expression of ANXA1and β-catenin in lung cancer tissues (r=0.638P=0.000).3.Western Blotting:①In both normal and tumor tissues, ANXA1and β-catenin proteins were expressed.②The amount of ANXA1protein in lung cancer tissue was significantly higher than the distal tissue.③The amount of P-catenin protein in lung cancer tissue was higher than the distal tissue.Conclusion:The amount of ANXA1, β-catenin mRNAs and proteins in lung cancer tissue showed a higher trend. The expressiones of the two genes in lung cancer tissues had significant correlation, and the differential expressiones of both in lung cancer were significantly correlated with tumor cells differentiation, tumor invasion and metastasis, suggesting that ANXA1gene might involved in the occurrence and development of NSCLC, which might affect the regulation of Wnt/β-Catenin signal pathway in the development of NSCLC. |
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