Title: Primary study on the effect and mechanism of celecoxib on the proliferation and cell cycle of SH-SY-5Y cells
Abstract: Objective To investigate the influence of high selective Cyclooxygenase-2 (COX-2) inhibitor celecoxib on proliferation and cell cycle of SH-SY-5Y cells and explore the potential COX-2dependent mechanism of celecoxib on the influence of SH-SY-5Y.Methods The SH-SY-5Y cells were treated by celecoxib with different doses(10 μmol/L,20 μmol/L,40μmol/L,80 μmol/L) for 48 hours.Cellular morphology was observed via inverted microscope.MethaberLzthiazuron (MTT) assay was used to evaluate proliferation of cells.The effect of celecoxib with different doses on cell cycle was detected by flow cytometry (10μmol/L,20 μmol/L,40 μmol/L,80 μmol/L).Western blot analysis for expression of CyclinD1,P16 and P21 was performed before and inoculation of celecoxib with different doses(10μmol/L,20μmol/L,40μmol/L,80μmol/L).Results After SH-SY-SY cells were treated by celecoxib with different doses (10 μmol/L,20 μmol/L,40 μmol/L,80 μmol/L) for 48 hours,the shape of SH-SY-5Y cells changed significantly.The shape of SH-SY-5Y cells pre-treated with celecoxib was in slender and fusiform shape.SH-SY-5Y cells became short or polygon after the treatment by celecoxib with concentrations.MTT assay showed the proliferation rate of SH-SY-5Y was 90.1%±3.6%,69.3% + 3.7% ,44.2% + 4.6% and 10.2% + 2.9% respectively with the dose of 10 μmol/L,20 μmol/L,40μmol/L and 80 μmol/L.Celecoxib inhibited the cells' proliferation in dose dependence.Flow cytometry showed the ratio of G0/G1 was 50.70% ± 4.0%,55.10%±5.2 %,67.20 %±3.9%,71.07 %±3.8% and 75.53%±4.8% respectively with the dose of 0μmol/L,10μmol/L,20 μmol/L,40 μmol/L and 80 μmol/L,in which the dose of 20 μmol/L,40 μmol/L and 80 μmol/L could significantly increase the ratio of G0/G1 compared with controls(P<0.01).The expression of CyclinDl,P21 and P16 were detected in SH-SY-SY cells.The western blot showed the expression level of CyclinD1 decreased with the increasing of the dose of celecoxib,while P21 and PI6 increased.Conclusions Celecoxib inhibits growth of SH-SY-5Y cells in G1 by down-regulating the expression of CyclinD1 and up-regulating the expression of P16 and P21 in vitro.Celecoxib would become a promising drug as an adjuvant therapy for children with neuroblastoma.
Key words:
Neuroblastoma; Cyclooxygenase 2; Cyclooxygenase 2 inhibitors; Cyclins
Publication Year: 2009
Publication Date: 2009-03-15
Language: en
Type: article
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