| Objective:1.To investigate the expressions of Semaphorin3A(SEMA3A)and Matrix Metalloproteinase 14(MMP-14)protein,observe their correlation and prognostic significance in non-small cell lung carcinoma(NSCLC).2.To explore the clinical significance and prognostive value of E-cadherin and vimentin,the markers of epithelial-mesenchymal transition(EMT)in NSCLC,and to study the correlations between SEMA3 Awith E-cadherin and vimentin.3.To explore the inhibitive effect of SEMA3 A on regulation of EMT in A549 cell.Methods:1.The study included 94 cases of NSCLC as experimental group and 80 cases of normal lung tissues as control group.The expressions of SEMA3A and MMP-14 protein by immunohistochemistry methods in two groups have been detected,and their correlation and prognostic value have been analyzed.2.The expression of E-cadherin and vimentin was measured by immunohistochemistry methods in NSCLC and the clinical significance,prognostic value and correlation with SEMA3A have been explored.3.A549 cells were cultured in vitro and divided into 3 groups:1)control;2)transforming growth factor-β1(TGF-β1);3)TGF-β1+SEMA3A.EMT was observed by inverted phase contrast microscope.Cell migration was detected by wound scratch assay.The expression of vimentin was measured by immunocytochemistrical staining,and the levels of E-cadherin and vimentin was detected by western blot.Results:1.The SEMA3A protein positive detection rates in NSCLC tissues were 36.17%,which were lower than those of the normal control that were 75.00%(P<0.05).The positive detection rate of SEMA3 A was found associated with the following phenomena no pleural invasion,no lymph mode metastasis,number of LN metastasis<4,positive rate of PCNA<25%,well and moderately differentiation,no vascular invasion and maximum diameter<5 cm(P<0.05).2.The MMP-14 protein positive detection rates in NSCLC tissues were 75.53%,which were higher than those of the normal control that were 25.00%(P<0.05).The positive detection rate of MMP-14 was found associated with the following phenomena:pleural invasion,lymph mode metastasis,number of LN metastasis>4,positive rate of PCNA>25%,poor differentiation and vascular invasion(P<0.05).3.There was a negative correlation between the protein expression of SEMA3A protein and MMP-14 in NSCLC(r=-0.852,P=0.000).Furthermore,it has been identified that the patients with low expression of SEMA3A and a higher expression of MMP-14 has worse disease prognosis.4.The E-cadherin protein positive detection rates in NSCLC tissues were 41.49%,which were lower than those of the normal control that were 77.50%(P<0.05).The positive detection rate of E-cadherin was found associated with the following phenomena:no pleural invasion,no lymph mode metastasis,number of LN metastasis<4,positive rate of PCNA<25%,well and moderately differentiation,no vascular invasion and maximum diameter<5cm(P<0.05).5.The vimentin protein positive detection rates in NSCLC tissues were 44.68%,which were higher than those of the normal control that were 7.50%(P<0.05).The positive detection rate of vimentin was found associated with the following phenomena:pleural invasion,lymph mode metastasis,number of LN metastasis>4,positive rate of PCNA>25%,poor differentiation and vascular invasion(P<0.05).6.There was a negative correlation between the protein expression of vimentin protein and E-cadherin in NSCLC(r=-0.793,P=0.000).And the expression of SEMA3A was positively correlated with the expression of E-cadherin(r=-0.857,P=0.000),but negatively correlated with the expression of vimentin(r=-0.785,P=0.000)in NSCLC.Furthermore,It has been identified that the patients with low expression of E-cadherin and a higher expression of vimentin has worse disease prognosis.7.TGF-β1-treated A549 cells in vitro gradually transformed from elliptic type to a spindle-like morphology with an vimentin positive muscle wire structure in cytoplasm.The migration rate in 48 h after treatment with TGF-β1 was increased 2.29 fold.The vimentin expression in TGF-β1 group increased to 6.86 fold and the expression of E-cadherin decreased by 5.70%compared with the control group.Pretreatment with SEMA3 Acould inhibited all this changes.Conclusions:1.This is the first report that the down-expression of SEMA3 A along with the loss of membranous E-cadherin,up-regulation of MMP-14 and vimentin are closely associated with NSCLC,which involved in pleural invasion,lymph mode metastasis,positive expression of PCNA,degree of differentiation and vascular invasion.Detecting the four protein levels can provide certain reference value in evaluating the metastasis,prognosis and malignant development monitoring of NSCLC.2.The results of the present study suggest that there is a novel mechanism of action for SEMA3A’s beneficial effect in NSCLC,which involves attenuation of EMT induced by TGF-β1 and cell migration. |
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