The Biological Pattern Of SEMA3B And Its Clinical Value In Pancreatic Carcinoma

Objective: Up-regulated Semaphorin3B(SEMA3B)cells were stablely constructed in pancreatic carcinoma(PC)CFPAC-1 and PANC-1 by using lentiviral vectors trancfection.The effect of SEMA3 B overexpression on cellular proliferation,invasion and migration of human PC cell lines was investigated in our study.Further,the correlation between the expression of SEMA3 B and clinicopathological pattern was analysed in patients with PC.Methods: The human PC CFPAC-1 and PANC-1 cell lines were culture in vitro,using the lentiviral vector mediated recombinant SEMA3 B transfected into PC cells.The cellular mRNA and protein were extracted to detect the expression of SEMA3 B by using qRT-PCR and western blotting,the transfection efficiency of SEMA3 B was identified.The effect of overexpression SEMA3 B on the proliferation of PC CFPAC-1 and PANC-1 cells was studied by using CCK-8 assay.The effect of overexpression SEMA3 B on cells invasion was detected by transwell assay.We also evaluated the effect of overexpression SEMA3 B on cells migration by wound healing assay.Further,31 cases of patients with PC were recruited from Jan.2009 to Jun.2016 in department of pathology of the first hospital Lanzhou University,and the related clinical data were analyzed.All specimens were identified as pancreatic carcinoma by pathologists and no other treatments such as radiotherapy and chemotherapy etc were performed preoperation.The expression level of SEMA3 B in 31 cases of PC was detected by immunohistochemistry.Fianlly,the correlation between SEMA3 B and the clinicopathological pattern of PC was discussed by ?2-test and Fisher's exact test.Results: The transfection efficiency of SEMA3 B in PC cells was detected by qRT-PCR and western blotting.The mRNA and protein level of SEMA3 B in transfected group was signifcantly higher than that in control group on two PC cell lines(P<0.001).The results of CCK-8 assay showed that the proliferation of transfected group was significantly inhibited by upregulated SEMA3B(P<0.001).The transwell assay showed that the number of penetrated cells in transfected group was significantly less than that in control group(P<0.001).The wound healing assay results showed that the scratch relative area of transfected group further increased in 24 h and 48 h was bigger than that in control group(P<0.001).In the 31 cases of PC,the expression of SEMA3 B presents downregulating in 25 cases and the other 6 cases presents upregulating.The expression level of SEMA3 B in PC tissues was closely related to the stage of tumo(P<0.05),local invasion(P<0.05),lymphatic metastasis(P<0.05)and distant metastasis(P<0.05).Conclusion: Lentiviral vector transfection can construct stable SEMA3 B upregulated model in PC CFPAC-1 and PANC-1 cell lines.Overexpression of SEMA3 B could significantly inhibit the proliferation,invasion and migration in PC CFPAC-1 and PANC-1 cells.The expression level of SEMA3 B in PC tissues was closely related to tumor stage,local invasion,lymphatic metastasis and distant metastasis.The lower of SEMA3 B,the higher degree of PC manignant.