Pyranocoumarin (±)-4′-O-Acetyl-3′-O-angeloyl-cis-khellactone Induces Mitochondrial-Dependent Apoptosis in HL-60 Cells

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-X_L. 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

ΔΨ_m:mitochondrial membrane potential

HPLC:high-performance liquid chromatography

NMR:nuclear magnetic resonance

4′-O-acetyl-3′-O-angeloyl-khellactone - Peucedanum praeruptorum - Umbelliferae - HL-60 - Apoptosis - mitogen-activated protein kinases - Bax, Bcl-2

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