Induction of Apoptosis by Apicularen A in Human Promyelocytic Leukemia Cell Line HL-60

Abstract

Apicularen A, a macrolide isolated from the myxobacterial genus Chondromyces, suppressed the proliferation of human promyelocytic leukemia cells (HL-60 cells), increased the release of lactate dehydrogenase and induced condensation and fragmentation of chromatin at 1 to 100 nM. In addition, it induced the DNA fragmentation, increased the percentage of annexin V-stained cells, and cleaved poly(ADP-ribose) polymerase (PARP), a substrate of caspase. In contrast, apicularen B, an N-acetylglucosamine glycoside of apicularen A, had no such effects at 100 nM. These findings indicated that apicularen A induces apoptosis in HL-60 cells by activating caspases. Phosphorylation of p44/42 MAPK, p38 MAPK and Akt was not induced by apicularen A at 100 nM, suggesting that the apicularen A-induced apoptosis in HL-60 cells is not regulated by the activation of p44/42 MAPK, p38 MAPK or Akt. Furthermore, by acridine orange staining of the cells, it was suggested that apicularen A but not apicularen B inhibits vacuolar-type H⁺-ATPase.

Key words

Apicularen A - apicularen B - Chondromyces - Polyangiaceae - Myxobacteriales - apoptosis - HL-60 cells - V-ATPase

References

• 1 Kunze B, Jansen R, Sasse F, Höfle G, Reichenbach H. Apicularens A and B, new cytostatic macrolides from Chondromyces species (myxobacteria): production, physico-chemical and biological properties.  J Antibiotics. 1998;  51 1075-80
• 2 Mesner P W, Budihardjo I, Kaufmann S H. Chemotherapy-induced apoptosis.  Adv Pharmacol. 1997;  41 461-99
• 3 Hong J J, Yamaki K, Ishihara K, Ahn J W, Zee O P, Ohuchi K. Induction of apoptosis of RAW 264.7 cells by the cytostatic macrolide apicularen A.  J Pharm Pharmacol. 2003;  55 1299-306
• 4 Roulston A, Reinhard C, Amiri P, Williams L T. Early activation of c-Jun N-terminal kinase and p38 kinase regulates cell survival in response to tumor necrosis factor α.  J Biol Chem. 1998;  273 10 232-9
• 5 Chin B Y, Petrache I, Choi A H, Choi M E. Transforming growth factor β1 rescues serum deprivation-induced apoptosis via the mitogen-activated protein kinase (MAPK) pathway in macrophages.  J Biol Chem. 1999;  274 11 362-8
• 6 Jun C D, Oh C D, Kwak H J, Pae H O, Yoo J C, Choi B M. et al . Overexpression of protein kinase C isoforms protects RAW 264.7 macrophages from nitric oxide-induced apoptosis: involvement of c-Jun N-terminal kinase/stress-activated protein kinase, p38 kinase, and CPP-32 protease pathways.  J Immunol. 1999;  162 3395-401
• 7 Koh J S, Lieberthal W, Heydrick S, Levine J S. Lysophosphatidic acid is a major serum noncytokine survival factor for murine macrophages which acts via the phosphatidylinositol 3-kinase signaling pathway.  J Clin Invest. 1998;  102 716-27
• 8 Jansen R, Kunze B, Reichenbach H, Höfle G. Apicularen A and B, cytotoxic 10-membered lactones with a novel mechanism of action from Chondromyces species (myxobacteria): Isolation, structure elucidation, and biosynthesis. Eur J Org Chem 2000: 913-9
• 9 Yoshimoto Y, Imoto M. Induction of EGF-dependent apoptosis by vacuolar-type H⁺-ATPase inhibitors in A431 cells overexpressing the EGF receptor.  Exp Cell Res. 2002;  279 118-27
• 10 Messmer U K, Briner V A, Pfeilschifter J. Tumor necrosis factor-alpha and lipopolysaccharide induce apoptotic cell death in bovine glomerular endothelial cells.  Kidney Int. 1999;  55 2322-37
• 11 Brunet A, Datta S R, Greenberg M E. Transcription-dependent and -independent control of neuronal survival by the PI3K-Akt signaling pathway.  Curr Opin Neurobiol. 2001;  11 297-305
• 12 Boyd M R, Farina C, Belfiore P, Gagliardi S, Kim J W, Hayakawa Y. et al . Discovery of a novel antitumor benzolactone enamide class that selectively inhibits mammalian vacuolar-type (H⁺)-ATPase.  J Pharm Exp Ther. 2001;  297 114-20
• 13 Bowman E J, Siebers A, Altendorf K. Bafilomycins: a class of inhibitors of membrane ATPases from microorganisms, animal cells, and plant cells.  Proc Natl Acad Sci USA. 1988;  85 7972-6
• 14 Dröse S, Bindseil K U, Bowman E J, Siebers A, Zeeck A, Altendorf K. Inhibitory effect of modified bafilomycins and concanamycins on P- and V-type adenosine-triphosphatases.  Biochemistry. 1993;  32 3902-6
• 15 Ohta T, Arakawa H, Futagami F, Fushida S, Kitagawa H, Kayahara M. et al . Bafilomycin A1 induces apoptosis in the human pancreatic cancer cell line Capan-1.  J Pathol. 1998;  185 324-30
• 16 Ishisaki A, Hashimoto S, Amagasa T, Nishihara T. Caspase-3 activation during the process of apoptosis induced by a vacuolar type H⁺-ATPase inhibitor.  Biol Cell. 1999;  91 507-13
• 17 Geisow M J, Beaven G H, Hart P D, Young M R. Site of action of a polyanion inhibitor of phagosome-lysosome fusion in cultured macrophages.  Exp Cell Res. 1980;  126 159-65
• 18 Moriyama Y, Nelson N. Lysosomal H⁺-translocating ATPase has a similar subunit structure to chromaffin granule H⁺-ATPase complex.  Biochim Biophys Acta. 1989;  980 241-7
• 19 Yoshimori T, Yamamoto A, Noriyama Y, Futai M, Tashiro Y. Bafilomycin A₁, a specific inhibitor of vacuolar-type H⁺-ATPase, inhibits acidification and protein degradation in lysosomes of cultured cells.  J Biol Chem. 1991;  266 17 707-12
• 20 Ishisaka A, Hashimoto S, Amagasa T, Nishihara T. Caspase-3 activation during the process of apoptosis induced by a vacuolar type H⁺-ATPase inhibitor.  Biol Cell. 1999;  91 507-13