The Mechanism Of Gemcitabine Induced High Secretion Of CXCL14 In Pancreatic Stellate Cells Which In Turn Promote The Invasion And Metastasis Of Pancreatic Cancer Cells

Objective In this research,we detected the expression of the CXCL14 in the supernatant of CaPSC which dealt with chemotherapeutic drug gemcitabine and the effect of the supernatant of CaPSC which high expression of CXCL14 on the proliferation and metastasis of pancreatic cancer cells and the related mechanism.Methods Obtain the CaPSC from the fresh pancreatic cancer tissue.The change of CXCL14 expression in the supernatant of CaPSC after dealt with gemcitabine was detected by ELISA.We constructed the CXCL14 overexpression plasmid and the interfering plasmid and then infect the CaPSC to obtain the stable transfection cell lines CaPSC-CXCL14 and CaPSC-shCXCL14.The effects of the supernatant of CaPSC-CXCL14 on proliferation,invasion and metastasis of pancreatic cancer cell line Panc-1 and BXPC-3 were detected by CCK-8,transwell and cell wound scratch assay.The changes of the expression of ?-catenin and EMT related m RNA and protein in Panc-1 and BXPC-3 cells were detected by real-time PCR and Western blot.Result The result of ELISA declared that CXCL14 was high expression in the supernatant of CaPSC after dealt with gemcitabine.We successfully constructed CXCL14 overexpression cell line CaPSC-CXCL14 and down-regulated expression cell line CaPSC-shCXCL14.The result of CCK-8 shown that after dealt with the supernatant of CaPSC-CXCL14,the proliferation rate of Panc-1 and BXPC-3 have no significant change compare with dealt with the supernatant of CaPSC(P>0.05).The result of wound healing assay and transwell assay shown that after dealt with the supernatant with CaPSC-CXCL14,the migration rate and invasive cell numbers of Panc-1 and BXPC-3 was significantly higher than that dealt with the supernatant of DMEM or CaPSC or CaPSC-shCXCL14(P<0.05 or P<0.01),and the effect was significant restrained by the XAV-939 which is the blocker of the ?-catenin passageway.The result of real-time PCR and Western blot shown that after dealt with the supernatant of CaPSC-CXCL14 the relate m RNA and protein of ?-catenin-EMT in Panc-1 and BXPC-3 was higher expression compare with dealt with the supernatant of DMEM or CaPSC or CaPSC-shCXCL14(P<0.05 or P<0.01)and the XAV-939 could significant restrained these effect.Conclusion This research demonstrated that the gemcitabine can significantly induce the high secretion of CXCL14 in pancreatic stellate cells and promote the invasion and metastasis of pancreatic cancer cells through the ?-catenin-EMT signaling pathway,but the effect on the proliferation of pancreatic cancer cells is not obvious.It is suggested that gemcitabine combined with the inhibitor of CXCL14 or ?-catenin-EMT pathway may be a new idea for pancreatic cancer treatment.