Expression Of C-Met In Tongue Squamous Cell Carcinoma And Its Role And Mechanism In Lymphangiogenesis And Lymphatic Metastasis

Objective:To investigate the expression of c-Met in tongue squamous cell carcinoma(TSCC)and analyze its relationship with clinicopathological features of patients,and to explore its role and mechanism in the process of lymphangiogenesis and lymphatic metastasis in TSCC.Methods:1.The pathological specimens of 58 patients with TSCC and 10 matched paracancerous tissue specimens that were surgically removed at the Tianjin Stomatological Hospital were studied.Immunohistochemistry was used to detect the expression of c-Met and VEGF-C protein.And lymphatic vessel density(LVD)was determined using specific lymphatic endothelial marker D2-40.The correlation of c-Met and VEGF-C expressions with the clinicopathologic characteristics of the patients and lymphangiogenesis were analyzed.2.c-Met specific ligand hepatocyte growth factor(HGF)and c-Met narrow-spectrum specific inhibitor of phosphorylation SU11274 were used to intervent the expression of c-Met in TSCC cell line CAL-27.The proliferation,migration and invasion of cells were detected by CCK-8 and Transwell assays,respectively.The expression of c-Met,p-Met,VEGF-C,E-cadherin,Vimentin,MMP9 and TIMP2 were detected by Western blot.Results:1.c-Met is highly expressed in TSCC.And its expression level in TSCC was significantly associated with the presence or absence of lymph node metastasis,clinical stage,T stage,and tumor differentiation(P<0.05).It was not related to the patient’s gender,age,smoking,alcohol consumption,and tumor location(P>0.05).VEGF-C is highly expressed in TSCC.The expression of VEGF-C in TSCC was significantly correlated with lymph node metastasis and clinical stages(P<0.05).It was not related to gender,age,smoking,alcohol consumption,tumor site,T stage,and tumor differentiation(P>0.05).The expression of c-Met and VEGF-C in TSCC was positively correlated(r=0.313,P=0.012).The LVD in lymph node metastasis of TSCC was significantly higher than that in the non-metastasis group(t=7.117,P=0.000),and the expression level of c-Met was significantly associated with LVD(t=3.483,P=0.000).The expression level of VEGF-C was significantly correlated with LVD(t=3.298,P=0.002).2.HGF significantly promoted the proliferation of CAL-27 cells(P<0.05).When the concentration of HGF is less than 30 ng/ml,the cell proliferation ability was enhanced with the increase of drug concentration in a dose-effect relationship;SU11274 inhibited the proliferation of CAL-27 cells in a dose and time dependent manner.HGF promoted cell migration and invasion(P<0.05);SU11274 inhibited cell migration and invasion(P<0.05).Western blot results showed that HGF promoted the expression of c-Met,p-Met,VEGF-C,Vimentin and MMP9,but inhibited the expression of E-cadherin and TIMP2;SU11274 inhibited the expression of p-Met,VEGF-C,Vimentin and MMP9.Expression,while promoted the expression of E-cadherin and TIMP2.When HGF and SU11274 co-operate,SU11274 can reverse the effects of HGF on cell proliferation,migration,invasion and the expression of related proteins.Conclusion:1.c-Met is closely related to the occurrence and development of TSCC,also related to its lymphangiogenesis and lymph node metastasis.The expression of c-Met is positively correlated with VEGF-C.c-Met may promote lymphangiogenesis and lymph node metastasis of TSCC by regulating VEGF-C.2.c-Met can influence cell migration and invasion by regulating CAL-27 epithelial mesenchymal transition and extracellular matrix degradation.