Induction of Apoptosis by 3,4′-Dimethoxy-5-hydroxystilbene in Human Promyeloid Leukemic HL-60 Cells

Abstract

3, 4′-Dimethoxy-5-hydroxystilbene (DMHS) is a hydroxystilbene compound obtained by methylation and acid hydrolysis of piceid (resveratrol-3-O-glucoside) from Polygonum cuspidatum. Herein, we report that DMHS induces programmed cell death or apoptosis in human promyelocytic leukemic HL-60 cells. We found that treatment of HL-60 cells with DMHS suppressed the cell growth in a concentration-dependent manner with an IC₅₀ value of 25 μM. DMHS increased internucleosomal DNA fragmentation in a time-dependent manner. The cell death by DMHS was partially prevented by the caspase inhibitor, zVAD-fmk. DMHS caused activation of caspases such as caspase-3, -8, and -9. Immunoblot experiments revealed that DMHS-induced apoptosis was associated with the induction of Bax expression. The release of cytochrome c from mitochondria into the cytosol was increased in response to DMHS. Taken together, our present results indicated that DMHS leads to apoptotic cell death in HL-60 cells through increased Bax expression and release of cytochrome c into cytosol and may be considered as a good candidate for a cancer chemopreventive agent in humans.

Hydroxystilbene - apoptosis - HL-60 cells - caspases - Bax - cytochrome c

References

• 1 Jang M, Cai L, Udeani G O, Slowing K V, Thomas C F, Beecher C WW, Fong H HS, Farnsworth N R, Kinghorn A D, Mehta R G, Moon R C, Pezzuto J M. Cancer chemopreventive activity of resveratrol, a natural product derived from grapes.  Science. 1997;  275 218-20
• 2 Subbaramaiah K, Chung W J, Michaluart P, Telang N, Tanabe T, Inoue H, Jang M, Pezzuto J M, Dannenberg A J. Resveratrol inhibits cyclooxygenase-2 transcription and activity in phorbol ester-treated human mammary epithelial cells.  Journal of Biological Chemistry. 1998;  273 21 875-82
• 3 Huang C, Ma W Y, Goranson A, Dong Z. Resveratrol suppresses cell transformation and induces apoptosis through a p53-dependent pathway.  Carcinogenesis. 1999;  20 237-42
• 4 Clement M V, Hirpara J L, Chawdhury S H, Pervaiz S. Chemopreventive agent resveratrol, a natural product derived from grapes, triggers CD95 signaling-dependent apoptosis in human tumor cells.  Blood. 1998;  92 996-1002
• 5 Tsan M -F, White J E, Maheshwari J G, Bremner T A, Sacco J. Resveratrol induces Fas signaling-independent apoptosis in THP-1 human monocytic leukemia cells.  British Journal of Haematology. 2000;  109 405-12
• 6 Wyllie A H. Apoptosis: an overview.  British Medical Bulletin. 1997;  53 451-65
• 7 Earnshaw W C, Martins L M, Kaufmann S H. Mammalian caspases: structure, activation, substrates, and functions during apoptosis.  Annual Review of Biochemistry. 1999;  68 383-424
• 8 Nagata S. Apoptosis by death factor.  Cell. 1997;  88 355-65
• 9 Ryu S Y, Han Y N, Han B H. Monoamine oxidase-a inhibitors from medicinal plants.  Archives of Pharmacentical Research. 1988;  11 230-9
• 10 Hsieh T, Juan G, Darzynkiewicz Z, Wu J M. Resveratrol increases nitric oxide synthase, induces accumulation of p53 and p21(WAF1/CIP1), and suppresses cultured bovine pulmonary artery endothelial cell proliferation by perturbing progression through S and G2.  Cancer Research. 1999;  59 2596-601
• 11 Chun Y J, Kim M Y, Guengerich F P. Resveratrol is a selective human cytochrome P450 1A1 inhibitor.  Biochemical and Biophysical Research Communications. 1999;  262 20-4
• 12 Ciolino H P, Yeh G C. Inhibition of aryl hydrocarbon-induced cytochrome P-450 1A1 enzyme activity and CYP1A1 expression by resveratrol.  Molecular Pharmacology. 1999;  56 760-7
• 13 Surh Y -J, Hurh Y -J, Kang J -Y, Lee E, Kong G, Lee S J. Resveratrol, an antioxidant present in red wine, induces apoptosis in human promyelocytic leukemia (HL-60) cells.  Cancer Letters. 1999;  140 1-10
• 14 Kang J J, Schaber M D, Srinivasula S M, Alnemri E S, Litwack G, Hall D J, Bjornsti M A. Cascades of mammalian caspase activation in the yeast Saccharomyces cerevisiae .  Journal of Biological Chemistry. 1999;  274 3189-98
• 15 Friesen C, Herr I, Krammer P H, Debatin K -M. Involvement of the CD95 (APO-1/FAS) receptor/ligand system in drug-induced apoptosis in leukemia cells.  Nature Medicine. 1996;  2 574-7
• 16 Krajewski S, Tanaka S, Takayama S, Schibler M J, Fenton W, Reed J C. Investigation of the subcellular distribution of the bcl-2 oncoprotein: residence in the nuclear envelope, endoplasmic reticulum, and outer mitochondrial membranes.  Cancer Research. 1993;  53 4701-14
• 17 Liu X, Kim C N, Yang J, Jemmerson R, Wang X. Induction of apoptotic program in cell-free extracts: requirement for dATP and cytochrome c.  Cell. 1996;  86 147-57
• 18 Hannun Y A, Obeid L M. Ceramide: an intracellular signal for apoptosis.  Trends in Biochemical Science. 1995;  20 73-7
• 19 Chun Y J, Ryu S Y, Jeong T C, Kim M Y. Mechanism-based inhibition of human cytochrome P450 1A1 by rhapontigenin.  Drug Metabolism and Disposition. 2001;  29 389-93