Apoptosis of U266 cells induced by brucine through death-receptor pathway / 白血病·淋巴瘤

ABSTRACT

Objective To investigate the mechanism of the apoptotic effect of brucine on human multiple myeloma. Methods MTT, morphology, flow cytometry, and RT-PCR were used to observe the apoptotic pathways of brucine on human multiple myeloma cell line-U266. Results The apoptotic effect of brucine showed a dose and time dependent manner, 48 hour IC50 0.16 mg/ml. The cells treated with brucine showed significant feature associated with apoptosis by Hoechst 33258 at fluorescence microscope.Mitochondrial membrance potential showed no statistical significant difference in different concentration with Rhodamine 123 by flow cytometry (P ＞0.05). The Caspase-3 expression detected by RT-PCR was increased at 12, 24 and 48 hours treated with brucine, and its gray value was (0.2597±-0.020), (0.5488±0.016), (0.6205±0.006), (0.6533±0.009) (P ＞ 0.05), detection of added z-IETD-fmk, z-LEHD-fmk (Caspase-8 and Caspase-9specific inhibitor) the expression of Caspase-3, the gray scale values were (0.7118±0.006), (0.2637±0.003)(P ＜0.01); and (0.7182±0.004) (0.7195±0.003) (P=0.836). Caspase-8 was activated. Conclusion Within the 0.4 mg/ml concentration brucine can induce apoptosis in U266 cells. Brucine induced apoptosis on cell line U266 through Caspase-8 activation by death-receptor pathway.