He Zhi-wen, Zhao Xiao-ying, Xu Rong-zhen, Wu Dong
Cancer Institute, The Second Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310009, China.
Zhejiang Da Xue Xue Bao Yi Xue Ban. 2006 Mar;35(2):209-14. doi: 10.3785/j.issn.1008-9292.2006.02.018.
To determine effects of berbamine on the growth of leukemia cell line NB4 and explore its possible mechanisms.
The growth of NB4 cells was examined with MTT assay. Morphological analysis and DNA agarose electrophoresis were used to detect apoptosis in NB4 cells, and the apoptosis rate was measured by flow cytometry. The PML/RAR alpha mRNA was determined by nested-PCR, and the Survivin mRNA was tested by RT-PCR. The expression of caspase 3 protein in NB4 cells was evaluated by flow cytometry.
The growth of NB4 cells was inhibited significantly after treated with berbamine at different concentrations for different time points, the IC(50)value was 3.860 microg/ml at 48 hours. Morphology analysis showed the characteristics of apoptosis, and the DNA agarose electrophoresis showed the typical DNA ladder. The apoptosis rate increased from 2.83% to 58.44% after treated with berbamine at 12 microg/ml for 48 hours. The expression of PML/RAR alpha mRNA presented no significant changes, however, Survivin mRNA was decreased dramatically. The protein expression of Caspase 3 increased significantly from 2.06% to 70.89% after treated with berberine at a concentration of 12 mug/ml for 48 hours.
Berbamine could inhibit the growth of leukemia cell line NB4. The induction of cell apoptosis may be one of the mechanisms for suppressing the growth of leukemia cell line NB4. Inhibition of Survivin mRNA and upregulation of Caspase 3 protein might be also involved in cell apoptosis.
探讨小檗胺对白血病细胞株NB4生长的影响及其可能机制。
采用MTT法检测NB4细胞的生长情况。用形态学分析和DNA琼脂糖凝胶电泳检测NB4细胞凋亡情况，并用流式细胞术检测凋亡率。采用巢式PCR法检测PML/RARα mRNA，用RT-PCR法检测Survivin mRNA。用流式细胞术评估NB4细胞中caspase 3蛋白的表达。
不同浓度小檗胺作用不同时间点后，NB4细胞生长受到显著抑制，48小时的IC(50)值为3.860μg/ml。形态学分析显示有凋亡特征，DNA琼脂糖凝胶电泳显示典型的DNA梯状条带。12μg/ml小檗胺作用48小时后，凋亡率从2.83%升至58.44%。PML/RARα mRNA表达无明显变化，但Survivin mRNA显著下降。12μg/ml黄连素作用48小时后，Caspase 3蛋白表达从2.06%显著升至70.89%。
小檗胺可抑制白血病细胞株NB4的生长。诱导细胞凋亡可能是其抑制白血病细胞株NB4生长的机制之一。抑制Survivin mRNA和上调Caspase 3蛋白也可能参与细胞凋亡过程。
