Expression Of CC Chemokine Receptor 7 And Vascular Endothelial Growth Factot -d In Lung Cancer

ObjectiveThe objective of this work is to investigate the expressions of CCR7 and VEGF-D protein in non small cell lung cancer and the relationship of their expressions with LVD and lymph node metastasis,and to reveal the molecular mechanism of CCR7-induced lymphangiogenesis and lymphy node metastasis in NSCLC.Materials and Methods1.Samples90 specimens of primary non small cell lung cancer were obtained from patients who had surgery in the First Affiliated hospital of China Medical University between 1980 and 2005,fixed in 10%paraformaldehyde,embedded in paraffin.2.Reagent The anti-human CCR7 goat monoclonal antibody and The anti-human VEGF-D rabbit monoclonal antibody were provide Santa Cruz company.The Ultro-sensitive S-Pkit and DAB agent kit were bought from Fujian Maxin Biological Company. pLNCX2-CCR7 plasmid were given as a present from Sam T.Hwang(National Institutes of Health,USA).3.MethodThe expressions of CCR7,VEGF-D and Lymphatic(D2-40 staining showed lymphatic) were detected in 90 specimens of human primary NSCLC by immunohistochemical SP method.Human lung big cell line BE1 cells were transfected by lipofection with CCR7 gene,the change of VEGF-D expression was observed by RT-PCR and Western blot.Correlation between the expression of CCR7 and VEGF-D was analyzed.4.Statistical analysisAll the data were analyzed with SPSS for Windows 13.0 software.The test was to compare the clinicopathological characteristics with the expression of CCR7 and VEGF-D.Spearman test was to the relationship of the expression of CCR7 and VEGF-D.The change of expression of VEGF-D circa transfection.and transfection. efficiency were computed by T-test.The result was expressioned by(?)±S,a=0.05 was standard,P＜0.05 had statistical significance.ResultImmunohistochemistry showed that CCR7 was distributed in cytoplasm and/or membrane of tumor cells,VEGF-D was distributed in cytoplasm of tumor cells.The expressions of CCR7,VEGF-D protein were found to be 70%(63/94) and 60%(54/94) in non small cell lung cancer,respectively,the expression of CCR7 and VEGF-D protein were closely related to the clinical stages(P=0.003)(P=0.005),LVD(P= 0.001;P＜0.001) and lymph node metastasis(P=0.004)(P=0.001) of non small cell lung cancer,but there was no correlation with age,gender,histology(P＞0.05). Furthermore,A significant correlation was found among CCR7 and VEGF-D(r=0.455, P＜0.001).In addition,the expression level of VEGF-D mRNA and protein increased in the transfected pLNCX2-CCR7 group(P＜0.05).ConclusionCCR7 could induce lymphangiogenesis and promote lymphy node metastasis by upregulating the expression of VEGF-D in NSCLC.