Effects And Related Mechanisms Of Human Umbilical Cord Mesenchymal Stem Cell Conditioned Medium On Proliferation,Migration And Invasion Of Pancreatic Cancer Cells

ObjectiveTo investigate the effects and related mechanisms of human umbilical cord mesenchymal stem cell conditioned medium(h UC-MSC-CM)on proliferation,migration and invasion of pancreatic cancer cells.MethodsMesenchymal Stem Cells(MSCs)were isolated from human umbilical cords.Then the MSCs were cultured,identified,and made into h UC-MSC-CM.Pancreatic cancer cell lines MIA-Pa Ca-2 and PANC1 were cultured in vitro.Pancreatic cancer cell lines treated with DMEM/F-12 medium containing 10% fetal bovine serum were the control group,and pancreatic cancer cell lines treated with h UC-MSC-CM containing 10% fetal bovine serum were h UC-MSC-CM group.Then MTT assay was used to detect cell proliferation ability,scratch assay was used to measure cell migration ability,and Transwell assay was used to detect invasion ability.RTQ-PCR assay was used to detect m RNA expression levels of Epithelial-Mesenchymal Transition(EMT)markers in cells.Western blot assays were used to detect the signaling pathways.1.The MTT assay showed that both MIA-Pa Ca-2 and PANC1 lines of h UC-MSC-CM group had significantly enhanced proliferation ability compared to the control group.The absorbance values were significantly different among the time points of 24,48 and 72 hours(***P<0.001).2.The scratch experiments showed that the relative scratch area healing rates at 48 h were(81.2 ± 2.2)% and(96.1 ± 4.2)% in the control and h UC-MSC-CM groups of MIA-Pa Ca-2 cells,respectively.The h UC-MSC-CM group was significantly higher than the control group(**P<0.01).The relative scratch area healing rates at 48 h were(67.4±1)% and(90.8±3.4)% in the control and h UC-MSC-CM groups of PANC1 cells,respectively.The h UC-MSC-CM group was significantly higher than the control group(***P<0.001).3.The Transwell assay showed that the number of cell invasion was(359±13)and(640±20)in the control and h UC-MSC-CM groups of MIA-Pa Ca-2 cells,respectively.The h UC-MSC-CM group was significantly higher than the control group(***P<0.001).The number of cell invasion was(100±7)and(790±32)in the control and h UC-MSC-CM groups of PANC1 cells,respectively.The h UC-MSC-CM group was significantly higher than the control group(***P<0.001)4.RTQ-PCR experiments showed that the m RNA expression levels of Snail and Vimentin were significantly upregulated and E-cadherin was significantly downregulated in the h UC-MSC-CM group of MIA-Pa Ca-2 and PANC1 compared with the control group(**P<0.01,***P<0.001).5.Western blot experiments showed that the expression levels of PI3 K and AKT were not significantly changed(P>0.05)and the expression levels of m TOR were significantly upregulated(*P<0.05,***P<0.001)in the h UC-MSC-CM group of both cell lines compared with the control group.Meanwhile,the expression levels of p-PI3 K,p-AKT,and p-m TOR wereResults significantly upregulated in the h UC-MSC-CM group of both cell lines compared with the control group(***P<0.001)ConclusionsThe h UC-MSC-CM may enhance the proliferation,migration and invasion of pancreatic cancer cells MIA-Pa Ca-2 and PANC1 by inducing EMT and activating PI3K/AKT/m TOR signaling pathway.