| Objective: Prostate cancer is one of the common cancers in older men with an increasing incidence. At present, incidence and mortality is increasing year by year. This disease depends on the presence of androgen, and most patients with early effective treatments of castration or combined androgen blockade, but after a few months to several years cancer is often transformed into hormone-refractory prostate cancer(HRPC). HRPC is one of the most complex issues in urological oncology, and the world there is no effective treatment now. Treatment of HRPC has become the bottleneck of Urology. Therefore, it is urgently needed that exploring more effective methods and treatments to improve the patient's prognosis and quality of life. At home and abroad studies in recent years have founded that COX-2 is high expression in a lot of tumors, and COX-2 played a key role in tumorigenesis and development. Nonsteroidal anti-inflammatory drugs(NSAIDs) especially the selective inhibitor of COX-2 have a wide range of tumor inhibition may reduce the incidence of tumors and prevent tumor formation, tumor recurrence and the transfer of the body. Celecoxib, is one of the selective inhibitor of COX-2 and it has been widely used in bone and joint diseases. In recent years, many reports show that it can induce anti-neoplastic activity on many human tumor cell lines. At present, rare has been elucidated on the influence of prostate cancer cells by inhibitor of COX-2. In this study, in order to investigate the possible antitumor mechanism of Celecoxib in COX-2 independent pathway, we observed the effect of selective COX-2 inhibitor Celecoxib on proliferation, apoptosis and cell cycle distribution in human prostate cancer PC-3 cells. Thus, theory evidence of Celecoxib in clinical application will be provided. Materials and method:Human prostate cancer PC-3 cells were cultivated in vitro and then treated with various concentrations of Celecoxib. Cell inhibitory growth rate was investigated by MTT assay,and the OD values were computed. Apoptosis rate, cell cycle distribution were detected by Flow cytometry(FCM). Using microscope observed the change of morphologic of PC-3 cells. Expression of Caspase-3 in PC-3 cells was detected by immunohistochemistry.The results were analyzed by SPSS13.0,and we established the standard of statistic significance asα=0.05.Results:1 Celecoxib on PC-3 cell growth inhibition: Cell inhibitory growth rate of PC-3 after treated with various concentrations of Celecoxib(40,80,120,160μmol/L) for 24 hours were 9.39%, 15.34%, 32.46%, 53.31% respectively. Cell inhibitory growth rate of PC-3 after treated with various concentrations of Celecoxib for 48 hours were 12.49%, 20.38%, 45.33%, 61.27% respectively. Cell inhibitory growth rate of PC-3 after treated with various concentrations of Celecoxib for 72 hours were 16.06%,21.43%,54.33%,81.60% respectively. Cell inhibitory growth rate in experiment groups after treated PC-3 cells for 24h, 48h, 72h were significantly different(P<0.05), at the same time, there was a significant difference between concentration groups and time groups(P<0.05).2 Celecoxib on cell cycle of PC-3: After PC-3 cells treated with various concentrations of Celecoxib(0,40,80,160μmol/L) for 48 hours,the cell percentage in G0/G1 phase of test groups were (26.78±0.89)%,(41.87±0.75)%,(49.25±0.68)%,(62.89±1.07)% respectively; the cell percentage in S phase were (51.86±1.60)%,(44.75±2.82)%,(35.48±1.88)%,(23.48±0.34)% respectively; the cell percentage in G2/M phase were (17.88±1.47)%,(15.65±1.53)%,(12.88±1.59)%,(10.88±0.93)% respectively. By one-way ANOVA, cell cycle distribution were significantly different among concentration groups, with the concentration of Celecoxib increased, cells in G0/G1 phase increased obviously and cells in S, G2/M phase decreased (P<0.05), the cells were blockaged on G0/G1 phase.3 Celecoxib on apoptosis of PC-3: Apoptosis cells increased significantly after PC-3 cells treated with Celecoxib(40,80,160μmol/L) for 48 hours. Apoptosis percentage(AP) were (12.04±1.64)%,(21.02±2.83)%,(32.68±3.64)% respectively; and AP in control (Celecoxib is 0μmol/L) was (4.86±0.78)%. By one-way ANOVA, AP were significantly different among concentration groups(P<0.05).4 Immunohistochemistry showed: The correspondent expression of Caspase-3 in PC-3 cells after treated with Celecoxib for 48 hours at the concentrations of 0μmol/L, 40μmol/L, 80μmol/L, 160μmol/L were (0.17±0.031),(0.23±0.042),(1.31±0.047),(1.39±0.071) respectively. The correspondent expression of Caspase-3 were significantly increased. The increasing expression of Caspase-3 existed significant difference among concentration groups(P<0.05).Conclusions:1 The survival of PC-3 cells was inhibited by Celecoxib in vitro with a concentration and time dependent manners.2 The number of PC-3 cells in G0/G1 phase was increased, while those in S and G2 phase was decreased by the effect of Celecoxib. Celecoxib induced cell cycle arrest in G0/G1 phase and inhibited the growth of PC-3 Cells.3 Celecoxib can induce apoptosis on PC-3 cells. With the Celecoxib concentration increased, apoptotic percentage of PC-3 cells increased in a concentration-dependent.4 The expression of Caspase-3 in PC-3 cells can be raised by Celecoxib in vitro with a concentrationdependent manner. So think that apoptosis in PC-3 cells can be increased by Celecoxib through the raised of Caspase-3 expression.
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