RhoGDI2 Regulates The Biolomcal Behaviors Of Pancreatic Cancer And Sensitivity To Gemcitabine

Background:Pancreatic cancer is one of the most malignant cancers in gastrointestinal tract,and gemcitabine is currently a first-line treatment for the disease.However,due to the existence of drug resistance,chemotherapy failure is frequently difficult to avoid.Rho GTPases as a superfamily of 20-30 members,plays an important role in cancer biological behaviors,with RhoGDI2(Rho GDP dissociation inhibitor2,RhoGDI2)as a key regulatory factor.Existing researches show that the ectopic expression of RhoGDI2 is relative to the migration and invasion capacities of pancreatic cancer cells.Moreover,it can also influence the chemotherapy resistance in other cancer cells.Purpose:In this research we aimed to identify the role of RhoGDI2 on pancreatic cancer cells' biological behaviors,investigate the correlation between the expression of RhoGDI2 and sensitivity of pancreatic cancer cells to gemcitabine,and detect the molecular mechanisms.Methods:1?Western blot was adopted to estimate the expression of RhoGDI2 in 5 pancreatic cancer cell lines(PANC-1?PATU8988?CFPAC-1?SW1990?PL45).2?siRNA(Small interfering RNA)was adopted to down-regulate RhoGDI2 in pancreatic cell lines,and overexpressed plasmid was adopted to up-regulate RhoGDI2 in pancreatic cell lines,then the efficiency of transfection was evaluated by western blot.After that,colony forming assay and CCK-8 kit were adopted to evaluate the proliferation of pancreatic cells,cell wound scratch assay was adopted to evaluate the migration of pancreatic cells and Trans well invasion assay was adopted to evaluate the invasion of pancreatic cells.To evaluate the apoptosis of pancreatic cells,Hoechst33342 staining solution was adopted,GreenNucTM Caspase-3 assay kit was adopted to find further molecular pathways.IC50?CCK-8 kit?Hoechst33342 staining solution and GreenNuc TM Caspase-3 assay kit were adopted to assess the sensibility of pancreatic cells to Gemcitabine.Results:Among 5 pancreatic cell lines(PANC-1?PATU8988?CFPAC-1?SW1990?PL45),RhoGDI2 was expressed highest in PATU8988 cells and lowest in PL45 cells.Western blot was adopted to confirm the transfection efficiency of RhoGDI2,and the differences between experiment groups and control groups were statistically significant.Cell function assay,including colony forming assay?CCK-8 kit?cell wound scratch assay and Transwell invasion assay,indicated that RhoGDI2 could obviously increase pancreatic cancer cells proliferation?migration and invasion.Hoechst33342 staining solution assay showed that RhoGDI2 could inhibit the apoptosis of pancreatic cancer cells,furthermore,GreenNucTM Caspase-3 staining solution assay shows that RhoGDI2 may promote apoptosis of pancreatic cancer cells via regulating caspase-3.The differences of IC50 of pancreatic cells for gemcitabine?CCK-8 kit?Hoechst33342 staining solution assay and GreenNucTM Caspase-3 staining solution assay indicated that RhoGDI2 could depress the pancreatic cancer cells sensitivity for gemcitabine.Conclusion:1?RhoGDI2 promotes the proliferation,migration and invasion of pancreatic cancer cells,inhibits the apoptosis of pancreatic cancer cells as well.2?RhoGDI2 can depress the pancreatic cancer cells sensitivity for gemcitabine.3?RhoGDI2 may depress the apoptosis of pancreatic cancer cells induced by gemcitabine via regulating caspase-3.