Studies on the Effects of Berbamine on the Human Leukemia Cell Line NB4.

Abstract

Purpose: To study the effects of berbamine on the human leukemia cell line NB4Methods: Berbamine compound was dissolved to the concentration of 1mg/ml with 0.9% sodium chloride and diluted to desired concentrations (as indicated in figure ledends 0.5,1,2,4,8,12,16ug/mL)with culture medium. Growth of NB4 cells was determined by MTT assay based upon the reduction of tetrazolium dye 3–4,5-dimethylthiazol-2,5-diphenyltetrazolium(MTT). 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.Result: 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 ug•mL-1at 48 hours. Typical nuclear condensation and apotosis body were observed as early as 24 hours following a 8μgμg•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 58.84% after treated with 12ug•mL-1 berbamine for 48 hours. The _expression of PML/RARα gene wasn't changed obviously.Conclusion: Berbamin could inhibit the proliferation of NB4 cells and inducing apoptosis might be its one of mechanisms. The berbamine-induced apoptosis is not related to the _expression of PML/RARα gene.