ABSTRACT
Objective To observe the anti-tumor effect on human multiple myeloma cell lines U266 and KM3 with a combination of varinostat and melphalan in vitro.Methods The cell proliferation of U266 and KM3 was datected with MTT assay when treated them with vorinostat alone and melphalan alone,then calculate their IC50 values respectively.Fixed the concentrations of vorinostator melphalan,the cell proliferation was datected in combination with melphalan/vorinostat in seriesly concentrations by MTr assay.Then to calculate drug combination index(CI).Results The IC50 value of U266 was 5.0-7.5 μmol/L and that of KM3 was 2.5-5.0 μmol/L when treated by vorinostat alone,the IC50 value of U266 was 40-60 μmol/L and that of KM3 was 60-80 μmol/L treated by melphalan alone.When fixed the concentration of vorinostat(in U266 the concentration was 1.25 μmol/L,in KM3 the concentration was 1.0 μ mol/L),Synergism(CI＜0.9)was observed when the concentrations of melphalan were 20 μmol/L,40 μmol/L,60 μ mol/L and 80 μ mol/L in U266,40 μmol/L,60 μmol/L,80 μmol/L and 100 μmol/L in KM3;When fixed the concertration of melphalan (in U266,was 10 μmol/L,in KM3 was 20 μmol/L),synergism(CI＜0.9)was observed when the concentrations of vorinostat were 1.0 μmol/L,2.5 μmol/L,5.0 μmol/L and 7.5 μmol/L in U266,and 1.0 μmol/L,2.5μmol/L,5.0 μmol/L in KM3.An additive effect was observed with the czombination of vorinostat 7.5 μmol/L plus melphalan 20 μmol/L in KM3(CI=0.93).Conclusion Vorinostat had potential anti-myeloma effect alone,and had synergistic anti-tumor effect with melphalan in vitro.
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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.
ABSTRACT
Objective:To investigate the effect of SU11248 proliferation and apoptosis of multiple myeloma cell line U266 in vitro and analyze its mechanisms.Methods:Effect of SU11248 on proliferation of U266 cells was detected by MTT assay.The ability of SU11248 to induce apoptosis of U266 cells was examined by cell cycle analysis,TUNEL and DNA fragmentation.Expression of c-myc,hTERT,Bcl-2 and Bax mRNA in U266 cells was assessed by RT-PCR analysis.Results:The proliferation of U266 cells was inhibited by SU11248 in dose-and time-dependent manners (P