The study on effects of Bcl-2 shRNA in enhancing methotrexatE-induced apoptosis and inhibiting cell proliferation in Raji cells / 肿瘤

ABSTRACT

Objective:To study the effect of Bcl-2 short hairpin RNA (shRNA) in enhancing methotrexate (MTX)-induced apoptosis of Raji cells. Methods:Expression plasmid containing Bcl-2 shRNA was transfected into Raji cells by lipofectmine 2000 and then the transfected cells were treated with MTX. The expression levels of Bcl-2 mRNA and protein were evaluated by RT-PCR and immunofluorescence method at 48 h of transfection. MTT assay was used to analyze cell proliferation at 24, 48 and 72 h. Apoptosis was detected by Giemsa staining and flow cytomertric cell cycle analysis. Results:After transfection with Bcl-2 shRNA, the expression levels of Bcl-2 mRNA and protein in Raji cells were significantly decreased (P<0.05). Bcl-2 shRNA transfection plus MTX treatment induced marked apoptosis, decreased in cell proliferation activity, and increased in apoptotic rate. The difference was significant compared with MTX group, negative shRNA plus MTX group, Bcl-2 shRNA group, and empty plasmid plus MTX group (P<0.05). Conclusion:Bcl-2 shRNA could enhance MTX-induced apoptosis and inhibition of cell proliferation in Raji cells.