Study Of BFGF Expression In Malignant Solid Tumors

Objective:To investigate expression of basic fibroblast growth factor (basic fibroblast growth factor, bFGF) in the clinical different types of solid tumors, and a preliminary discussion bFGF expression and tumor cells, growth, differentiation and correlation of clinical staging of lymph node metastasis. By understanding how tumors bFGF expression levels, bFGF monoclonal antibody in clinical targeting therapy could provide a basis for the application.Methods:1. Preparation of bFGFmAb producing from ascites:Hehybridomas cells of secreting bFGF monoclonal antibodies, which were cryopreservated in our lab, were recoveried and cultured, BALB/c mice were immunized by injecting incomplete Freund's adjuvant(IFA) intraperitoneally. Intraperitoneal injection of hybridoma cells in mouse 2×106/mice,7～12 day ascites, extract ascites, which containing the bFGFmAb was produced. bFGFmAb was purified from the ascites through the Protein G-Sepharose affinity column, Indirect method ESLIA was used to detect the bFGFmAb titration before and aftter purification. The concentration of purified bFGFmAb stock solution was assayed by BCA standard solution. The bFGFmAb purity was determined by SDS-PAGE electrophoresis. The bFGFmAb solution was cryopreservated at-20℃for use.2. Immunohistochemical SP Assay was used to detect 131 cases bFGF protein expression in paraffin-embedded tissues of primary non-small cell lung cancer, breast cancer, colon cancer and melanoma.Results1.26.5ml ascites containing bFGFmAb was prepared, the bFGFmAb titration was 1:1024000, while the titration of purified bFGFmAb solution was 1:4096000. The result of SDS-PAGE electrophoresis revealed the high purity of purified bFGFmAb solution. The Concentration of purified bFGFmAb solution was 8.586mg/ml, the total weight of bFGFmAb was 89.148mg.2. bFGF expression of SP testing result was as below:1) The positive bFGF overexpression of non-small cell lung cancer (56.25%). breast cancer (48.72%), colorectal cancer (53.66%), melanoma (52.61%).2) Among different pathological types of NSCLC, adenocarcinoma bFGF overexpression (56.25%), squamous cell carcinoma bFGF expression (64.26%) The difference was not obvious. In different differentiation degree of NSCLC, low differentiated carcinoma (70%), middle to high differentiation (33.33%), there were significant differences. StageⅠNSCLC of bFGF expression (28.57%), stageⅡbFGF expression (44.44%), stageⅢandⅣbFGF expression (75%), there were significantly difference.3) bFGF positive expression in lymph node metastasis in breast cancer (61.54%) no lymph node metastasis(30.77%), statistically significant difference between the two groups(p=0.036); bFGF positive expression in stageⅠBreast cancer (20.07%),Ⅱ(57.89%),Ⅲ～Ⅳ(71.43%), statistically significant differences among the three groups(P=0.041).4) bFGF positive expression in high differentiation adenocarcinoma (71.43%), low differentiated adenocarcinoma (35.00%), statistically significant differences between the two groups (P=0.032); stageⅠColon bFGF Positive expression (27.27%),Ⅱstage (28.57%),Ⅲ～Ⅳstage (73.91%). statistically significant differences among the three groups(P=0.013).bFGF expression positive no lymph node metastasis in 5 cases (27.78%), lymph node metastasis in 17 cases (73.91%), the comparison between the two groups was statistically significant differences (P=0.003).5) bFGF Positive expression in stageⅠmelanoma (25.00%),Ⅱ(66.67%),Ⅲ～Ⅳ(50.00%), statistically significant differences among the three groups(P=0.046). bFGF positive expression in melanoma in lymph node metastasis in 8 cases (72.73%), no lymph node metastasis in 2 cases (25.00%), the comparison between the two group was statistically significant difference (P=0.040);Conclusions:1, bFGF expression in lung adenocarcinoma and squamous cell carcinoma, breast cancer, colon cancer, melanoma and other malignant tumor cells was expressed in the cytoplasm.2, bFGF expression was related in tumor tissues and clinical TNM staging of malignant tumor cell differentiation, and lymph node metastasis.3,bFGF could become malignant degree of cancer diagnosis, differences between tumor cells, predicting malignant tumor targets.