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Objective:The aim of this study was to study the effect of Bcl-2 small hairpin RNA(shRNA) enhancing methotrexate(MTX)-induced apoptosis of Raji cells.Methods:Expression plasmid with Bcl-2 shRNA was transfected into Raji cells by Lipofectmine 2000 and then treated with MTX.At 48 h of transfection,the expression level of Bcl-2 mRNA and protein was evaluated by RT-PCR and immunofluorescence.MTT assay was used to analyze cell proliferation at 24,48 and 72 h.Apoptosis was detected by Giemsa staining and flow cytometric cell cycle analysis.Results:After transfection with Bcl-2 shRNA,the expression levels of Bcl-2 mRNA and protein in Raji cells decreased(P < 0.05).Using Giemsa staining,cells transfected with Bcl-2 shRNA combined with MTX at 48 h displayed changes of apoptosis.MTX significantly inhibited the growth of cells after transfected with Bcl-2 shRNA(P < 0.05).Apoptotic rates of the Raji cells treated with Bcl-2 shRNA combined with MTX significantly increased(P < 0.05),compared with either control shRNA/MTX combination or MTX-treatment cells alone.Conclusion:Our results suggest the shRNA against Bcl-2 mRNA could increase MTX-induced apoptosis of Raji cells.
Objective: The aim of this study was to study the effect of Bcl-2 small hairpin RNA (shRNA) enhancing methotrexate (MTX) -induced apoptosis of Raji cells. Methods: Expression plasmid with Bcl-2 shRNA was transfected into Raji cells by Lipofectmine 2000 and then treated with MTX. At 48 h of transfection, the expression level of Bcl-2 mRNA and protein was evaluated by RT-PCR and immunofluorescence. MTT assay was used to analyze cell proliferation at 24, 48 and 72 h.Apoptosis was detected by Giemsa staining and flow cytometric cell cycle analysis. Results: After transfection with Bcl-2 shRNA, the expression levels of Bcl-2 mRNA and protein in Raji cells decreased (P <0.05). Using Giemsa staining, cells transfected with Bcl- 2 shRNA combined with MTX at 48 h displayed changes of apoptosis. MTX significantly inhibited the growth of cells after transfected with Bcl-2 shRNA (P <0.05) .Apoptotic rates of the Raji cells treated with Bcl-2 shRNA combined with MTX increased increased (P <0.05), compared with ei ther control shRNA / MTX combination or MTX-treatment cells alone. Confluence: Our results suggest the shRNA against Bcl-2 mRNA could increase MTX-induced apoptosis of Raji cells.