Abstract
[Objective] To investigate the effect of berbamine (BBM) on the proliferation and apoptosis of retinoblastoma (RB) HXO-RB44 cells and its possible mechanism in vitro.[Methods] RB cells in logarithmic growth phase were divided into BBM treated group and control group.RB cells in BBM treated group were cultured with different concentrations of BBM (2,4,8,16 and 32 mg/L) for 24,48 and 72 hours,respectively.The proliferation was assayed by methyl Thiazolyl tetrazolium (MTT).RB cells were cultured with different concentrations of BBM (4,8 and 16 mg/L) for 24 hours.The early apoptotic rates were detected by flow cytometry; the expression of bcl-2 and Bax were measured by enzyme-linked immunosorbent assay (ELISA) and the activity of Caspase-3 was detected by colorimetric assay.[Results] BBM could obviously inhibit the proliferation of RB cells in a time-and dose-dependent manner (24 hours:F=70.547,P＜0.01; 48 hours:F=603.438,P＜0.01; 72 hours:F=577.521,P＜0.01).The 1C50value at 24,48 and 72 hours were 25.26,10.94 and 6.25 mg/L,respectively.Necrosis rates of control group andBBM treated group were (1.25±0.45)％,(4.10±2.95)％,(4.39±0.21)％ and (10.54±4.38) ％ respectively; the difference between two groups was statistically significant (F =6.527,P＜0.05).Apoptotic and necrosis rates in advanced stage of control group and BBM treated group were (2.13±0.71)％,(5.45 ± 2.31)％,(9.86 ± 3.18)％ and ( 11.10 ± 1.70)％,respectively.The difference between two groups was statistically significant (F =10.845,P＜0.05).Early apoptotic rates of control group andBBM treated group were (0.51±0.26)％,(2.68±0.35)％,(5.97±0.50)％ and (11.22±1.17) ％,respectively.The difference between two groups was statistically significant (F=144.976,P＜0.01).In addition,BBM dose-dependently reduced bcl-2 level and increased Bax expression,causing the reduction of the bcl-2/Bax protein ratio as well as increased the Caspase-3 activity in RB cells remarkably (bcl2:F=835.726,P＜0.01; bax:F=111.963,P＜0.01; Caspase-3:F=298.058,P＜0.01).[Conclusion]s BBM can inhibit the proliferation and induce apoptosis or necrosis of RB cells in vitro,downregulating the expression of bcl-2,up-regulating the expression of Bax.Along with increased Caspase-3activity these may be the apoptotic mechanisms. Key words: Retinoblastoma/drug therapy; Berbamine/therapeutic use; Apoptosis; Cell proliferation
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