小檗胺对NB4白血病细胞生长的影响

Studies on Effects of Berbamine on Human Leukemia Cell Line NB4

目的:探讨小檗胺对 NB4白血病细胞生长的影响及其机制。方法:体外培养 NB4人白血病细胞,经不同浓度小檗胺处理不同时间后,MIT 法检测药物对 NB4细胞增殖的抑制作用;细胞形态学观察和 DNA 琼脂糖电泳检测细胞凋亡;流式细胞仪检测 DNA 含量及周期;巢式 PCR 检测 PML/RARα融合基因表达。结果:NB4细胞经不同浓度小檗胺处理不同时间后细胞生长受抑,并呈时间剂量依赖性,48 h IC_(50)为3.86μg·mL^(-1);细胞形态学观察到细胞凋亡特征,DNA 琼脂糖电泳显示典型的凋亡梯形图,经12μg·mL^(-1)的小檗胺处理48 h 后流式细胞仪检测发现细胞凋亡率明显增加,从2.83%增加58.44%;但未发现药物作用后 NB4细胞 PML/RARα基因表达水平的改变。结论:小檗胺能抑制 NB4细胞生长,其机制之一可能是诱导细胞凋亡。

Objective： To study the effects of berbamine on the human leukemia cell line NB4. Methods： Berbamine compound was dissolved to the concentration of 1 mg · mL^-l with 0. 9% sodium chloride and diluted to desired concentrations with culture medium. MTT was used to examine the effect of berbamine on growth of NB4 cells. Morphological observation was used to detect the characteristic changes of apoptosis in NB4 cells and DNA agarose electrophoresis were used to detect the characteristic changes of apoptosis in NB4 cells, while apoptosis rate was measured by flow cytometry assay. The expression of PML/RARα gene was determined by nested - PCR. Results： MTT results showed berbamine could inhibit the growth of NB4 cells significantly after treated with different concentration of berbamine for different time, IC50 was 3. 86 μg · mL^-1 at 48 hours. Typical nuclear condensation and apotosis body were observed as early as 24 hours following a 8 μg · mL^-1 berbamine treatment;while the DNA agarose electrophoresis also show the typical DNA ladder,and the apoptosis rate of NB4 cell was increased from 2. 83% to 58.44% after treated with 12 μg · mL^-1 berbamine for 48 hours. The expression of PML/RARα gene wasn＇ t changed obviously. Conclusion：Berbamine can inhibit the proliferation of NB4 cells and inducing apoptosis might be its one of mechanisms.