The Study Of Relationship Between Cyclooxygenase-2, Matrix Metalloproteinase 14 And Proliferation, Invasion And Metastasis In Pancreatic Cancer Cells

Objective: By measuring the impact of different concentrations of cyclooxygenase-2 inhibitor celecoxib on proliferation, invasion, migration and COX-2, MMP-14 expression in pancreatic cancer cells, investigate the effect of COX-2 inhibitor celecoxib on pancreatic cancer and the related mechanism.Methods: 1. Human pancreatic cancer cell line PANC-1 cells were cultured in vitro; 2. PANC-1 cells were treated with diverse concentrations of celecoxib(0、20、60、100μmol/L) for different durations(24、48、72h). Cell proliferation, invasion and migration capabilities were measured by MTT colorimetry, transwell invasion assay, and scratch assay separately. At the same time, the protein expression of COX-2 and MMP-14 was assessed by Enzyme-linked immunosorbent assay(ELISA); 3. Statistical analysis of data.Results: 1. The capabilities of proliferation in PANC-1 cells were reduced in a concentration- and time-dependent manner after treated with different concentration of celecoxib for 24, 48 and 72 h by MTT colorimetry assay. This inhibitory effect was intensified along with the elevated concentrations of celecoxib and the prolongation of action time(P<0.05); 2. The capabilities of invasion in PANC-1 cells were decreased in a concentration-dependent manner after treated with different concentration of celecoxib for 24 h by transwell invasion assay, the number of cells invading through matrigel was 300.654±12.558, 271.041±10.569, 184.003±10.865 and 92.667±7.567, respectively(F=237.182, P<0.05); 3. The capabilities of migration in PANC-1 cells were impaired in a concentration-dependent manner after treated with different concentration of celecoxib for 24 h by scratch assay, the relative migration distance was 11.400±0.959, 9.511±1.110, 5.856±0.778 and 2.844±0.416, respectively(F=59.516, P<0.05); 4. The normal morphology of PANC-1 cells were spindle or polygonal in a good growth condition,but this morphology was changed after treated with different concentration of celecoxib for 24 h by cell morphological observation. Compared with normal cells, PANC-1 became round and shrinking followed by dead cells; 5. The protein expression of COX-2 and MMP-14 in PANC-1 cells was correspondingly reduced in a concentration-dependent manner after treated with different concentration of celecoxib for 24 h by ELISA, the protein expression of COX-2 was 0.876±0.023, 0.682±0.027, 0.505 ± 0.022 and 0.384 ± 0.019, respectively(F=262.537, P<0.05), the protein expression of MMP-14 was 7.955±0.297, 7.031±0.178, 6.072±0.209 and 5.047±0.216, respectively(F=89.247, P<0.05). 6. The protein expression of MMP-14 was positively correlated with the protein expression of COX-2 in PANC-1 cells(r=0.873,P<0.01).Conclusion: 1. COX-2 inhibitor celecoxib attenuates the proliferation of human pancreatic cancer cell line PANC-1 in a concentration- and time-dependent manner, the invasion and migration in a concentration-dependent manner; 2. COX-2 inhibitor celecoxib may down-regulate MMP-14 expression via inhibiting COX-2 expression, then attenuating the proliferation、invasion and migration of PANC-1 cells in a concentration-dependent manner, contributing to its anti-tumor effect in pancreatic cancer.