Reduced Expression Of SERPINB6 Gene In CNE-1 Cells Effects Of Proliferation,Invasion And Migration

Objective:The purpose of this study is to investigate the effect of SERPINB6 gene expression on proliferation,invasion and migration of CNE-1 cells and the activity of MMP-2,a metastatic protein of nasopharyngeal carcinoma,by interfering with the expression of SERPINB6 gene in CNE-1 cells.To further reveal the pathogenesis of nasopharyngeal carcinoma,provide theoretical basis for clinical diagnosis,and provide experimental basis for gene therapy of nasopharyngeal carcinoma with SERPINB6 as a potential target.Methods:1.T he human nasopharyngeal carcinoma CNE-1 cell line was used as the research object for culture.First,the pre-experiment was carried out The pre-experiment was divided into five groups:mock group(non-transfected,group A),N.C-siRNA group(transfected negtive control siRNA,group B),siRNA-SERPINB6-1 control group(transfected with siRNA-SERPINB6-1,group C),siRNA-SERPINB6-2 control group(transfected with siRNA-SERPINB6-2,roup D),siRNA-SERPINB6-3 control group(transfected with siRNA-SERPINB6-3,group E).2.The transfection rate(NC-FAM)of CNE-1 cells was detected by SERPINB6 small interferences using a common siRNA economical package.RT-PCR was used to detect the expression of SERPINB6 in five groups of cells,and the best interference group was selected as a follow-up formal experiment.3.The formal experiment was divided into three groups:mock group(group A),N.C-siRNA group(group B)and siRNA-SERPINB6 group(experimental group C,the best interference).MTT assay,Transwell invasion test and scratch test were used to detect the effects of SERPINB6 on proliferation,invasion and migration of nasopharyngeal carcinoma CNE-1 cells.4.Detection of MMP-2 expression at protein level by Western Blot.Results:1.In the preliminary experiment,after transfection of siRNA,the relative expression of SERPINNB6 in the experimental group was significantly lower than that in the negative control group and the blank group(P<0.05).The transfection of siRNA3 sequence,that is,experimental group C,decreased most significantly(P<0.01),and the interference effect was the best,which indicated that the experiment successfully interfered with SERPINB6,so the subsequent experiment adopted the siRNA3 sequence of experimental group C.2.The proliferation rates of the si-RNA group were 0.184(+0.017),0.278(+0.019)and 0.389(+0.023)by MTT every 24 hours,which were significantly higher than those of the negative control group and the blank group.(p<0.05).3.Scratch test showed that the migration distance of cells in the experimental group transfected with si-RNA was significantly faster than that in the negative control group and the blank group.(p<0.05).4.Transwell test showed that the number of cells entering the lower compartment in the si-RNA group was significantly higher than that in the negative control group and the blank group,and the invasive ability was enhanced.(p<0.05).5.Western Blot method showed that the level of MMP-2 protein activation in the si-RNA group was significantly higher than that in the negative control group and the blank group(p<0.05).Conclusion:1.SERPINB6 can inhibit cell proliferation,migration and invasion,which can provide a new target for gene therapy.2.Exogenous transfection of si-SERPINB6 promotes the migration and invasion of CNE-1 cells,and the expression of MMP-2 increases.SERPINB6 may be involved in the metastasis regulation of CNE-1 cells by mediating MMPS activation.