Establishment Of GPSM2 Eukaryotic Expression Vector And The Effects To Migration Ability Of Human Pancreatic Cancer

ObjectiveTo build a stable cell line with the G-protein signaling modulator 2(GPSM2)high expression,and study the relationship between GPSM2 and migration ability of human pancreatic cancer.To explore the use of GPSM2 as a molecular target for the treatment of pancreatic cancer to provide new ideas and strategies.Methods1.To construct GPSM2 over-expressed plasmid,transfect human pancreatic cancer MIA-PaCa-2 cells.2.Human pancreatic cancer MIA-PaCa-2 cells were transfected with pCMV-Tag 3B-GPSM2(GPSM2 transfection group)or empty pCMV-Tag 3B vectors(negative control group),with untreated MIA-PaCa-2 cells as blank control.In each group of cells,the GPSM2 mRNA expressions were measured by RT-PCR.3.In each group of cells,the protein expressions of GPSM2 and β-catenin were determined by Western-blot analysis.4.The changes of ability of migration were examined by Transwell.Results1.The recombinant cell line with high expression of GPSM2 was successfully constructed.2.The results of RT-PCR showed that the expression of GPSM2 mRNA was significantly high when compared with negative control group and blank control group(all P<0.01).Compared with the blank control group,the efficiency of GPSM2 gene was 73.3 times.There was no significant difference in the expression of GPSM2 mRNA between negative control group and blank control group.(P>0.05).3.Western-blot results showed that there were high expression of GPSM2 and β-catenin in GPSM2 transfection group than negative control group and blank control group(all P<0.05).There was no statistical difference in the expression of GPSM2 and P-catenin between negative control group and blank control group(all P>0.05).In addition,a positive linear relationship existed between GPSM2 and P-catenin expressions in pancreatic cancer cells(P<0.05).4.In GPSM2 transfection group compared with negative control group and blank control group,transwell results showed that the number of migrated cells was significantly increased(all P<0.01).ConclusionsMigrated cells count of the recombinant cell line with high expression of GPSM2 was significantly increased.The upregulation of GPSM2 expression in human pancreatic cancer cells can increase the expression of β-catenin protein,which may promote the migration of human pancreatic cancer cells.