The Effects Of 1, 25(OH)₂D₃ Associated With Celecoxib On The Proliferate Inhibition And Apoptosis Induction In Breast Cancer Cell Line Hs578T

Objective To study the effect of 1,25-dihydroxyvitamin D₃ (VitD₃) associated with celecoxib(CELE) on growth and apoptosis in breast cancer cell line Hs578T . Methods The cell numbers were compared by using MTT method, The cell cycle and apoptosis percentage were analyzed by flow cytometric, quantitatively apoptosis cells were determinaed by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) and HE staining.The expression of bcl-2,Bax,Ki-67,VEGF,survivin,COX2 and VDR were examined by using immunohistochemistry S-P method in different doses combined with Image pro plus 6.0 analyse software . Results 1)Incubation with 10-7mol/L VitD₃ associated with 5×10^-5mol/L CELE or 10^-8 mol/L VitD₃ associated with 5×10^-5mol/L CELE,Hs578T cells exhibited significant growth inhibition, The ratio of apoptosis were 53.0% and 48.0%.2)Flow cytometric analysis indicated cell′s S or G0/G1 arrest along with increasing apoptotic peak and percentage.3)The AI(apoptosis index) as detected by TUNEL stain in 10^-7mol/L VitD₃ associated with 5×10^-5mol/L CELE or 10^-8 mol/L VitD₃ associated with 5×10^-5mol/L CELE were higher than that in the negative control.4)Treatment of Hs578T cells with VitD₃ associated with CELE resulted in concentration-dependent decreases in bcl-2 and VEGF and increased in Bax proteins. Conclusion 1)10^-7mol/L VitD₃ associated with 5×10^-5mol/L CELE or 10-8 mol/L VitD₃ associated with 5×10^-5mol/L CELE exhibited obvious inhibitory effects on the cell proliferation.Down-regulation of Ki-67 and VEGF expression in Hs578T cells might be one of its possible mechanisms.2) 10^-7mol/L VitD₃ associated with 5×10^-5mol/L CELE or 10^-8 mol/L VitD3 associated with 5×10^-5mol/L CELE could obviously induce apoptosis of Hs578T cells in vivo.Enhancing Baxexpression and inhibiting Bcl-2 expression in Hs578T cells might be its mechanisms.3) Flow cytometric analysis indicated cell′s S or G0/G1 arrest along with increasing apoptotic peak and percentage.