Berberine and a Berberis lycium extract inactivate Cdc25A and induce alpha-tubulin acetylation that correlate with HL-60 cell cycle inhibition and apoptosis.
Berberis lycium Royle (Berberidacea) from Pakistan and its alkaloids berberine and palmatine have been reported to possess beneficial pharmacological properties. In the present study, the anti-neoplastic activities of different B. lycium root extracts and the major constituting alkaloids, berberine and palmatine were investigated in p53-deficient HL-60 cells. The strongest growth inhibitory and pro-apoptotic effects were found in the n-butanol (BuOH) extract followed by the ethyl acetate (EtOAc)-, and the water (H(2)O) extract. The chemical composition of the BuOH extract was analyzed by TLC and quantified by HPLC. 11.1 microg BuOH extract (that was gained from 1mg dried root) contained 2.0 microg berberine and 0.3 microg/ml palmatine. 1.2 microg/ml berberine inhibited cell proliferation significantly, while 0.5 microg/ml palmatine had no effect. Berberine and the BuOH extract caused accumulation of HL-60 cells in S-phase. This was preceded by a strong activation of Chk2, phosphorylation and degradation of Cdc25A, and the subsequent inactivation of Cdc2 (CDK1). Furthermore, berberine and the extract inhibited the expression of the proto-oncogene cyclin D1. Berberine and the BuOH extract induced the acetylation of alpha-tubulin and this correlated with the induction of apoptosis. The data demonstrate that berberine is a potent anti-neoplastic compound that acts via anti-proliferative and pro-apoptotic mechanisms independent of genotoxicity.
Khan
Musa
M
Department of Plant Sciences, Quaid-i-Azam University, Islamabad, Pakistan.
Giessrigl
Benedikt
B
Vonach
Caroline
C
Madlener
Sibylle
S
Prinz
Sonja
S
Herbaceck
Irene
I
Hölzl
Christine
C
Bauer
Sabine
S
Viola
Katharina
K
Mikulits
Wolfgang
W
Quereshi
Rizwana Aleem
RA
Knasmüller
Siegfried
S
Grusch
Michael
M
Kopp
Brigitte
B
Krupitza
Georg
G
D000107:Acetylation; D017209:Apoptosis / Q000187:drug effects*; D001599:Berberine / Q000494:pharmacology*; D028142:Berberis / Q000737:chemistry*; D015153:Blotting, Western; D002453:Cell Cycle / Q000187:drug effects*; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D020552:Comet Assay; D018922:HL-60 Cells; D006801:Humans; D010766:Phosphorylation / Q000187:drug effects; D010936:Plant Extracts / Q000494:pharmacology*; D018517:Plant Roots / Q000737:chemistry; D000090063:Proto-Oncogene Mas; D014404:Tubulin / Q000378:metabolism*; D020687:cdc25 Phosphatases / Q000037:antagonists & inhibitors* / Q000378:metabolism
C000715470:MAS1 protein, human; D010936:Plant Extracts; D000090063:Proto-Oncogene Mas; D014404:Tubulin; D001599:Berberine; C517004:CDC25A protein, human; D020687:cdc25 Phosphatases