Abstract
The pyranocoumarin (±)-4′-O-acetyl-3′-O-angeloyl-cis-khellactone (PC) isolated from Radix Peucedani (root of Peucedanum praeruptorum Dunn) showed a dose-dependent effect at 10 - 30 μg/mL on causing apoptotic DNA and
nuclear fragmentations in HL-60 cells. After 24 h of PC treatment there were losses
of mitochondrial membrane potential and cytochrome c. PC also increased total cellular and mitochondrial Bax protein, stimulated an increase
in caspase-dependent Bcl-2 cleavage but showed no effect on Bcl-XL. These observations strongly suggest activation of the mitochondria apoptotic pathway.
The pan-specific caspase inhibitor, ZVAD-fmk, abolished the PC-induced apoptosis,
whereas the caspase-8 inhibitor IETD-fmk showed no effect, implying the involvement
of the caspase 9 pathway. PC caused a 2 to 12 hour transient increase in phospho-ERK,
and a 72 h-long activation of JNK. Pre-treatment with the MEK inhibitor PD98059, which
suppresses ERK activation, paradoxically promoted PC-induced mitochondrial cytochrome
c release, procaspase-3 and -8 cleavage, and enhanced apoptosis. Our results show that
PC triggers mitochondria-mediated apoptosis in HL-60 cells, and the involvement of
ERK and JNK signal pathways in the process.
Abbreviations
PC:(±)-4′-O-acetyl-3′-O-angeloyl-cis-khellactone
MAPK:mitogen-activated protein kinase
ERK:extracellular signal-regulated kinase
MEK:MAPK/ERK kinase
ΔΨm:mitochondrial membrane potential
HPLC:high-performance liquid chromatography
NMR:nuclear magnetic resonance
Key words
4′-O-acetyl-3′-O-angeloyl-khellactone -
Peucedanum praeruptorum
- Umbelliferae - HL-60 - Apoptosis - mitogen-activated protein kinases - Bax, Bcl-2
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Wang-Fun Fong
Department of Biology and Chemistry
City University of Hong Kong
Kowloon
Hong Kong SAR
P. R. China
Phone: +852-2788-9181
Fax: +852-2788-7406
Email: bhwffong@cityu.edu.hk