Effect Of SAE2 On Epithelial Interepithelial Transformation Of Bile Duct Carcinoma Cells

Objective: To investigate the effect of SAE2 gene on the epithelial mesenchymal transition(EMT)of cholangiocarcinoma cells by investigating the effects of SAE2 gene overexpression and SAE2 gene interference on the cell proliferation the relationship between apoptosis and cell invasiveness of cholangiocarcinoma cells.Methods:Transfection of cholangiocarcinoma cell line by liposome transfection,the recombinant eukaryotic expression plasmid of SAE2 gene was constructed,and SAE2 cell line was stably expressed.They were divided into the following 3 groups: SAE2 overexpression group,knockdown SAE2 group and empty plasmid group.By real-time RT-PCR and Western blot detection of SAE2 mRNA and protein expression in each group.MTT assay was used to detect the proliferation of cells before and after transfection.The apoptosis of each group was detected by FCM,and the transwell assay was performed by Transwell assay changes in tumor cell invasiveness.Results: The results of RT-PCR and WB showed that SAE2 mRNA and protein expression was higher in SAE2 overexpression group than in knockdown SAE2 group,which was higher than that of empty plasmid group(P^0.05).Knockdown SAE2 cells,The expression of SAE2 mRNA and protein was significantly lower than the empty plasmid group,the difference was statistically significant(P ^ 0.05).MTT results showed that there was no significant difference in MTT between groups(P < 0.05).On the 2nd day,the MTT OD value in SAE2 overexpression group was higher than that in knockdown SAE2 group,which was higher than that of empty plasmid group.Statistically significant(P^0.05).At 3 days,the MTT OD of SAE2 overexpression group cells was higher than that of knockdown SAE2 group,which was higher than that of empty plasmid group(P^0.05).The differences the two groups have were statistically significant(P < 0).05).The OD value was significantly higher than that of the SAE2 knockdown group(P^0.05).The results of FCM showed that the percentage of cell count in Annexin V positive results was not statistically significant(P^0.05)in each group ofcells at 1d,and the percentage of cell count in Annexin V positive results in SAE2 overexpression group was lower than that at 2d.The SAE2 group was lower than the empty plasmid group,The differences the two groups have were statistically significant(P < 0).05).The percentage of cells that knocked down the SAE2 group of Annexin V positive results was not statistically different from that of the empty plasmid group(P^0.05).The percentage of cells with Annexin V positive cell count was lower in SAE2 overexpression group than in knockdown SAE2 group,which was lower than that in empty plasmid group.The percentage of cell count in Annexin V positive result in empty plasmid group was lower than that in SAE2 knockdown group.Significance(P^0.05).The results of Transwell invasion assay showed that there was no significant difference in cell invasiveness between groups(P^0.05).At 2 days,the invasiveness of SAE2 overexpression group was higher than that of knockdown SAE2 group,higher than that of empty plasmid group.The differences the two groups have were statistically significant(P < 0).05).There was no significant difference in the invasiveness between the SAE2 group and the empty plasmid group(P^0.05).At 3 days,the invasiveness of the SAE2 overexpression group was higher than the knockdown.The SAE2 group was higher than the empty plasmid group.The invasiveness of the empty plasmid group was higher than that of the knockdown SAE2 group,and the difference was statistically significant(P^0.05).Conclusion: In this study,QBC939 cell line stably expressing SAE2 gene was successfully constructed.It was found that QBC939 cells highly expressing SAE2 had stronger proliferative ability and invasion ability than QE939 cells that knocked down SAE2,and the apoptosis was weaker than that of QC939 cells knocked down by SAE2.SAE2 may be an important regulatory gene for proliferation,apoptosis and invasion of QBC939 cells.