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Aim:To investigate the effects of small interfering RNA of cyclooxygenase-2(COX-2) on the proliferation and apoptosis of human multiple myeloma RPMI8226cells and its relation with the Bcl-2 family in vitro.Methods:Transcription andexpression of COX-2 in human myeloma RPMI8226 cells were checked by RT-PCRand Western blot analysis,respectively.The COX-2 siRNA fragment targetingexon 5 of COX-2 gene was transfected into the cells with the Amaxa nucleofectiontechnique.The inhibition of cell growth was detected by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium (MTT) assay.Apoptosis was estimatedby Annexin-V/propidium iodide double-labeled cytometry and confirmed by termi-nal deoxynucleotidyl transferase-mediated dUTP nick end-labeling assay.Bcl-2and Bax expression was evaluated by Western blot analysis.Results:The COX-2siRNA fragment could be successfully transfected into RPMI8226 cells,whichresulted in the significant inhibition of transcription and expression of COX-2 inthe myeloma cells.Proliferation of the transfected cells was inhibited andapoptosis was induced (6.52%±0.32%,12.53%±2.52%,24.39%±3.51% and36.48%±4.96% for 0,12,24,and 48 h,respectively) in a time-dependent manner(P<0.01).However,the expression of Bcl-2 and Bax in the RPMI8226 cells had nosignificant changes after nucleofection.Conclusion:COX-2 siRNA transfectioncan suppress COX-2 expression in human myeloma RPMI8226 cells,which leadsto growth inhibition and apoptosis independent of Bcl-2.
Aim: To investigate the effects of small interfering RNA of cyclooxygenase-2 (COX-2) on the proliferation and apoptosis of human multiple myeloma RPMI 8226 cells and its relation with the Bcl-2 family in vitro. Methods: Transcription andexpression of COX-2 in human myeloma RPMI8226 cells were checked by RT-PCR and Western blot analysis, respectively. The COX-2 siRNA fragment targettingexon 5 of COX-2 gene was transfected into the cells with the Amaxa nucleofectiontechnique. The inhibition of cell growth was detected by 3- ( 4,5-dimethyl-2-thiazolyl) -2,5-diphenyl-2H-tetrazolium (MTT) assay. Apoptosis was estimated by Annexin-V / propidium iodide double-labeled cytometry and confirmed by termi-nal deoxynucleotidyl transferase-mediated dUTP nick end-labeling assay. Bcl-2 and Bax expression was evaluated by Western blot analysis. Results: The COX-2 siRNA fragment could be successfully transfected into RPMI8226 cells, whichresulted in the significant inhibition of transcription and expression of COX-2 inthe myelo ma cells. Proliferation of the transfected cells was inhibited and apoptosis was induced (6.52% ± 0.32%, 12.53% ± 2.52%, 24.39% ± 3.51% and36.48% ± 4.96% for 0,12,24, and 48 h, respectively However, the expression of Bcl-2 and Bax in the RPMI8226 cells had nosignificant changes after nucleofection. Confluence: COX-2 siRNA transfectioncan suppress COX-2 expression in human myeloma RPMI8226 cells , which leadsto growth inhibition and apoptosis independent of Bcl-2.