Apoptosis of human leukemia HL-60 cells and murine leukemia WEHI-3 cells induced by berberine through the activation of caspase-3.
The aim of this study was to clarify the mechanisms of apoptosis, cytotoxicity, DNA damage and fragmentation, as well as the production of reactive oxygen species (ROS) and Ca+2, induced by berberine in human promyelocytic leukemia HL-60 and murine myelomonocytic leukemia WEHI-3 cells. The levels of Bcl-2 and Bax, the changes of mitochondria membrane potential (MMP), cytochrome c release and activation of caspase-3 were also investigated in both cell lines. The flow cytometry and DAPI staining assays indicated that berberine induced cytotoxicity in both cell lines examined. Flow cytometry assay also showed that berberine induced ROS and Ca+2 production, decreased the levels of MMP and increased the activity of caspase-3 in both cell lines examined. Berberine-induced apoptosis was accompanied by increased levels of Ca+2 and a decrease in the mitochondrial membrane potential, leading to the release of cytochrome c and the cleavage of pro-caspase-3. Western blotting also showed that berberine increased the levels of Bax and cytochrome c and decreased the levels of Bcl-2 in both cell lines. Inhibition of caspase-3 activation (z-VAD-fmk: cell-permeable broad-spectrum caspase inhibitor) completely blocked berberine-induced apoptosis in both HL-60 and WEHI-3 cells. Therefore, berberine induced apoptosis in both examined cell lines through the activation of caspase-3.
Lin
Chin-Chung
CC
Graduate Institute of Chinese Medical Science, School of Chinese Medicine, Taiwan, ROC.
Kao
Shung-Te
ST
Chen
Guang-Wei
GW
Ho
Heng-Chien
HC
Chung
Jing-Gung
JG
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001599:Berberine / Q000494:pharmacology*; D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism; D053148:Caspase 3; D061945:Caspase Inhibitors; D020169:Caspases / Q000378:metabolism*; D002470:Cell Survival / Q000187:drug effects; D045304:Cytochromes c / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects; D005456:Fluorescent Dyes / Q000737:chemistry; D018922:HL-60 Cells; D006801:Humans; D007211:Indoles / Q000737:chemistry; D007425:Intracellular Membranes / Q000187:drug effects / Q000502:physiology; D007951:Leukemia, Myeloid / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D015473:Leukemia, Promyelocytic, Acute / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D051379:Mice; D008928:Mitochondria / Q000187:drug effects / Q000502:physiology; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000096:biosynthesis / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D051028:bcl-2-Associated X Protein / Q000378:metabolism; D051025:bcl-Associated Death Protein / Q000378:metabolism
D061945:Caspase Inhibitors; D005456:Fluorescent Dyes; D007211:Indoles; D019253:Proto-Oncogene Proteins c-bcl-2; D017382:Reactive Oxygen Species; D051028:bcl-2-Associated X Protein; D051025:bcl-Associated Death Protein; D001599:Berberine; C007293:DAPI; D045304:Cytochromes c; C505407:CASP3 protein, human; C505409:Casp3 protein, mouse; D053148:Caspase 3; D020169:Caspases; D002118:Calcium