Preferences help
enabled [disable] Abstract
Number of results
2011 | 58 | 1 | 111-117
Article title

Antitumor activity of antimicrobial peptides against U937 histiocytic cell line

Title variants
Languages of publication
We investigated cytotoxic activity of antimicrobial peptides of different origin (both naturally occurring and synthetic), structure and known mechanisms of action against human histiocytic lymphoma cell line U937. The strongest cytotoxic activity against U937 cell line was shown by Pexiganan MSI-78, followed by Citropin 1.1, Protegrin 1 and a synthetic lipopeptide, N-α-palmitoyl-l-lysyl-l-lysine amide (Pal-Lys-Lys-NH2). The cytotoxic activity of the peptides was more dependent on the time of incubation than concentration. Only for the lipopeptide, whose mode of action was restricted to disruption of electric potential of the cell membrane, the correlation between cytotoxicity and concentration was almost linear. The high cytotoxicity of Pexiganan MSI-78, Protegrin 1 and the lipopeptide could be basically explained by their membranolytic activity leading to necrosis. However, in the case of Citropin 1.1, the cell membrane integrity was disrupted only slightly and independently of the peptide concentration. Therefore, some other mechanism of action might be responsible for its strong dose-dependent cytotoxic activity, e.g., membranolytic activity leading to apoptosis. Furthermore, TNF-α production due to LPS (lipopolysaccharide) stimulation was suppressed by the presence of Citropin 1.1, Pexiganan MSI-78 or Protegrin 1, but not by Buforin 2 or the lipopeptide. Our experiments have shown that cytotoxic activity is not limited to some specific molecular structure of a peptide, but rather to the length of the peptide chain as it is likely to affect the efficiency of the tumor cell membrane disruption and interaction with LPS.
Physical description
  • Department of Cell Biology, Faculty of Medical Biotechnology, Medical University of Gdansk, Gdańsk, Poland
  • Faculty of Chemistry, University of Gdansk, Gdańsk, Poland
  • Department of Cell Biology, Faculty of Medical Biotechnology, Medical University of Gdansk, Gdańsk, Poland
  • Department of Inorganic Chemistry, Faculty of Pharmacy, Medical University of Gdansk, Gdańsk, Poland
  • Department of Cell Biology, Faculty of Medical Biotechnology, Medical University of Gdansk, Gdańsk, Poland
  • Aisenbrey C, Bechinger B, Gröbner G (2008) Macromolecular crowding at membrane interfaces: adsorption and alignment of membrane peptides. J Mol Biol 375: 376-385.
  • Ambroggio EE, Separovic F, Bowie JH, Fidelio GD, Bagatolli LA (2005) Direct visualization of membrane leakage induced by the antibiotic peptides: maculatin, citropin, and aurein. Biophys J 89: 1874-1881.
  • Avrahami D, Shai YA (2004) A new group of antifungal and antibacterial lipopeptides derived from non-membrane active peptides conjugated to palmitic acid. J Biol Chem 279: 12277-12285.
  • Beisswenger C, Bals R (2005) Functions of antimicrobial peptides in host defense and immunity. Curr Protein Pept Sci 6: 255-264.
  • Bessler WG, Cox M, Lex A, Suhr B, Wiesmüller KH, Jung G (1985) Synthetic lipopeptide analogs of bacterial lipoprotein are potent polyclonal activators for murine B lymphocytes. J Immunol 135: 1900-1905.
  • Boman HG, Agerberth B, Boman A (1993) Mechanisms of action on Escherichia coli of cecropin P1 and PR-39, two antibacterial peptides from pig intestine. Infect Immun 61: 2978-2984.
  • Bradshaw JP (2003) Cationic antimicrobial peptides. issues for potential clinical use. Biodrugs 17: 233-240.
  • Bulet P, Stöcklin R, Menin L (2004) Anti-microbial peptides: from invertebrates to vertebrates. Immunol Rev 198: 169-184.
  • Carraway KL 3rd, Funes M, Workman HC, Sweeney C (2007) Contribution of membrane mucins to tumor progression through modulation of cellular growth signaling pathways. Curr Top Dev Biol 78: 1-22.
  • Chen F-Y, Lee M-T, Huang HW (2003) Evidence for membrane thinning effect as the mechanism for peptide-induced pore formation. Biophys J 84: 3751-3758.
  • Chen Q, Wade D, Kurosaka K, Wang ZY, Oppenheim JJ, Yang D (2004) Temporin A and related frog antimicrobial peptides use formyl peptide receptor-like 1 as a receptor to chemoattract phagocytes. J Immunol 173: 2652-2659.
  • Cho JH, Sung BH, Kim SC (2009) Buforins: Histone H2A-derived antimicrobial peptides from toad stomach. Biochim Biophys Acta 1788: 1564-1569.
  • Cruciani RA, Barker JL, Zasloff M, Chen HC, Colamonici O (1991) Antibiotic magainins exert cytolytic activity against transformed cell lines through channel formation. Proc Natl Acad Sci USA 88: 3792-3796.
  • Cudic M, Otvos Jr L (2002) Intracellular targets of antibacterial peptides. Curr Drug Targets 3: 101-106.
  • de Planque MRR, Bonev BB, Demmers JAA, Greathouse DV, Koeppe RE 2nd, Separovic F, Watts A, Killian JA (2003) Interfacial anchor properties of tryptophan residues in transmembrane peptides can dominate over hydrophobic matching effects in peptide-lipid interactions. Biochemistry 42: 5341-5348.
  • Dubin A, Mak P, Dubin G, Rzychon M, Stec-Niemczyk J, Wladyka B, Maziarka K, Chmiel D (2005) New generation of peptide antibiotics. Acta Biochim Pol 52: 633-638.
  • Epand RM, Vogel HJ (1999) Diversity of antimicrobial peptides and their mechanisms of action. Biochim Biophys Acta 1462: 11-28.
  • Esper DH, Harb WA (2005) The cancer cachexia syndrome: a review of metabolic and clinical manifestations. Nutr Clin Pract 20: 369-376.
  • Fernandez DI, Gehman JD, Separovic F (2009) Membrane interactions of antimicrobial peptides from Australian frogs. Biochim Biophys Acta 1788: 1630-1638.
  • Ferrara N, Kerbel RS (2005) Angiogenesis as a therapeutic target. Nature 438: 967-974.
  • Fields GB, Noble RL (1990) Solid phase peptide synthesis utilizing 9-fluorenylmethoxycarbonyl amino acids. Int J Pept Protein Res 35: 161-214.
  • Gottesman MM (2002) Mechanisms of cancer drug resistance. Ann Rev Medi 53: 615-627.
  • Gottler LM, de la Salud Bea R, Shelburne CE, Ramamoorthy A, Marsh EN (2008) Using fluorous amino acids to probe the effects of changing hydrophobicity on the physical and biological properties of the β-hairpin antimicrobial peptide protegrin-1.Biochemistry 47: 9243-9250.
  • Gottler LM, Ramamoorthy A (2009) Structure, membrane orientation, mechanism, and function of pexiganan - A highly potent antimicrobial peptide designed from magainin. Biochim Biophys Acta 1788: 1680-1686.
  • Harmey JH, Bucana CD, Lu W, Byrne AM, McDonnell S, Lynch C, Bouchier-Hayes D, Dong Z (2002) Lipopolysaccharide-induced metastatic growth is associated with increased angiogenesis, vascular permeability and tumor cell invasion. Int J Cancer 101: 415-422.
  • Heinzelmann M, Mercer-Jones MA, Flodgaard H, Miller FN (1998) Heparin-binding protein (CAP37) is internalized in monocytes and increases LPS-induced monocyte activation. J Immunol 160: 5530-5536.
  • Isaacson RE (2003) MBI-226. Micrologix/Fujisawa. Curr Opin Investig Drugs 4: 999-1003.
  • Jacob L, Zasloff M (1994) Potential therapeutic applications of magainins and other antimicrobial agents of animal origin. Ciba Found Symp 186: 197-216.
  • Jang WS, Li XS, Sun JN, Edgerton M (2008) The P-113 fragment of histatin 5 requires a specific peptide sequence for intracellular translocation in Candida albicans, which is independent of cell wall binding. Antimicrob Agents Chemother 52: 497-504.
  • Kamysz W, Kochańska B, Kędzia A, Ochocińska J, Maćkiewicz Z, Kupryszewski G (2002) Statherin SV2 and its analogue. Synthesis and evaluation of antimicrobial activity. Pol J Chem 76: 801-806.
  • Kamysz W, Silvestri C, Cirioni O, Giacometti A, Licci A, Della Vittoria A, Okroj M, Scalise G (2007) In vitro activities of the lipopeptides palmitoyl (Pal)-Lys-Lys-NH2 and Pal-Lys-Lys alone and in combination with antimicrobial agents against multiresistant gram-positive cocci. Antimicrob Agents Chemother 51: 354-358.
  • Kobayashi S, Chikushi A, Tougu S, Imura Y, Nishida M, Yano Y, Matsuzaki K (2004) Membrane translocation mechanism of the antimicrobial peptide buforin 2. Biochemistry 43: 15610-15616.
  • Koczulla AR, Bals R (2003) Antimicrobial peptides: current status and therapeutic potential. Drugs 63: 389-406.
  • Kokryakov VN, Harwig SS, Panyutich EA, Shevchenko AA, Aleshina GM, Shamova OV, Korneva HA, Lehrer RI (1993) Protegrins: leukocyte antimicrobial peptides that combine features of corticostatic defensins and tachyplesins. FEBS Lett 327: 231-236.
  • Lehrer RI, Ganz T (1999) Antimicrobial peptides in mammalian and insect host defense. Curr Opin Immunol 11: 23-27.
  • Mader JS, Richardson A, Salsman J, Top D, de Antueno R, Duncan R, Hoskin DW (2007) Bovine lactoferricin causes apoptosis in Jurkat T-leukemia cells by sequential permeabilization of the cell membrane and targeting of mitochondria. Exp Cell Res 313: 2634-2650.
  • Mader JS, Hoskin DW (2006) Cationic antimicrobial peptides as novel cytotoxic agents for cancer treatment. Expert Opin Investig Drugs 15: 933-946.
  • Mander T, Hill S, Hughes A, Rawlins P, Clark C, Gammon G, Foxwell B, Moore M (1997) Differential effects on TNF alpha production by pharmacological agents with varying molecular sites of action. Int J Immunopharmacol 19: 451-462.
  • Mani R, Cady SD, Tang M, Waring AJ, Lehrer RI, Hong M (2006) Membrane-dependent oligomeric structure and pore formation of a beta-hairpin antimicrobial peptide in lipid bilayers from solid-state NMR. Proc Natl Acad Sci 103: 16242-16247.
  • Matsuzaki K, Sugishita K, Ishibe N, Ueha M, Nakata S, Miyajima K, Epand RM (1998) Relationship of membrane curvature to the formation of pores by magainin 2. Biochemistry 37: 11856-11863.
  • Melo MN, Castanho MARB (2007) Omiganan interaction with bacterial membranes and cell wall models. Assigning a biological role to saturation. Biochim Biophys Acta 1768: 1277-1290.
  • Moore AJ, Devine DA, Bibby MC (1994) Preliminary experimental anticancer activity of cecropins. Pept Res 7: 265-269.
  • Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65: 55-63.
  • Naumov GN, Townson JL, MacDonald IC, Wilson SM, Bramwell VH, Groom AC, Chambers AF (2003) Ineffectiveness of doxorubicin treatment on solitary dormant mammary carcinoma cells or late-developing metastases. Breast Cancer Res Treat 82: 199-206.
  • Oren Z, Shai Y (1998) Mode of action of linear amphipathic alpha-helical antimicrobial peptides. Biopolymers 47: 451-463.
  • Papo N, Shai Y (2003) Exploring peptide membrane interaction using surface plasmon resonance: differentiation between pore formation versus membrane disruption by lytic peptides. Biochemistry 42: 458-466.
  • Paquette DW, Waters GS, Stefanidou VL, Lawrence HP, Fridcn PM, O 'Connor SM, Sperati JD, Oppenheim FG, Butchens LH, Williams RC (1997) Inhibition of experimental gingivitis in beagle dogs with topical salivary histatins. J Clin Periodontol 24: 216-222.
  • Park CB, Kim MS, Kim SC (1996) A novel antimicrobial peptide from Bufo bufo gargarizans. Biochem Biophys Res Commun 218: 408-413.
  • Park CB, Kim HS, Kim SC (1998) Mechanism of action of the antimicrobial peptide buforin II: buforin II kills microorganisms by penetrating the cell membrane and inhibiting cellular functions. Biochem Biophys Res Commun 244: 253-257.
  • Polozov IV, Polozova AI, Tytler EM, Anantharamaiah GM, Segrest JP, Woolley GA, Epand RM (1997) Role of lipids in the permeabilization of membranes by class L amphipathic helical peptides. Biochemistry 36: 9237-9245.
  • Reddy KVR, Yedery RD, Aranha C (2004) Antimicrobial peptides: premises and promises. Int J Antimicrob Agents 24: 536-547.
  • Rosenfeld Y, Papo N, Shai Y (2006) Endotoxin (lipopolysaccharide) neutralization by innate immunity host-defense peptides. J Biol Chem 281: 1636-1643.
  • Shai Y (1999) Mechanism of the binding, insertional destabilization of phospholipid bilayer membranes by helical antimicrobial and cell non-selective membrane lytic peptides. Biochim Biophys Acta 1462: 55-70.
  • Simmaco M, Mignogna G, Canofeni S, Miele R, Mangoni ML, Barra D (1996) Temporins, antimicrobial peptides from the European red frog Rana temporaria. Eur J Biochem 242: 788-792.
  • Smiley S, Almyroudis N, Segal BH (2005) Epidemiology and management of opportunistic infections in immunocompromised patients with cancer. Abstr Hematol Oncol 8: 20-30.
  • Sundstrom C, Nilsson K (1976) Establishment and characterization of a human histiocytic lymphoma cell line (U937). Int J Cancer 17: 565-577.
  • Takabayashi T, Takahashi N, Okamoto M, Yagi H, Sato M, Fujieda S (2009) Lipopolysaccharides increase the amount of CXCR4, and modulate the morphology and invasive activity of oral cancer cells in a CXCL12-dependent manner. Oral Oncol 45: 968-973.
  • Utsugi T, Schroit AJ, Connor J, Bucana CD, Fidler IJ (1991) Elevated expression of phosphatidylserine in the outer membrane leaflet of human tumor cells and recognition by activated human blood monocytes. Cancer Res 51: 3062-3066.
  • von Haussen J, Koczulla R, Shaykhiev R, Herr C, Pinkenburg O, Reimer D, Wiewrodt R, Biesterfeld S, Aigner A, Czubayko F, Bals R (2008) The host defence peptide LL-37/hCAP-18 is a growth factor for lung cancer cells. Lung Cancer 59: 12-23.
  • Wabnitz PA, Bowie JH, Wallace JC, Tyler MJ (1999) The citropin peptides from the skin glands of the Australian Blue Mountains tree frog Litoria citropa. Part 2: sequence determination using electrospray mass spectrometry. Rapid Commun Mass Spectrom 13: 1724-1732.
  • Wallach D (1984) Preparations of lymphotoxin induce resistance to their own cytotoxic effect. J Immunol 132: 2464-2469.
  • Wegener KL, Wabnitz PA, Carver JA, Bowie JH, Chia BC, Wallace JC, Tyler MJ (1999) Host defence peptides from the skin glands of the Australian blue mountains tree-frog Litoria citropa. Solution structure of the antibacterial peptide citropin 1.1. Eur J Biochem 265: 627-637.
  • Wu M, Maier E, Benz R, Hancock RE (1999) Mechanism of interaction of different classes of cationic antimicrobial peptides with planar bilayers and with the cytoplasmic membrane of Escherichia coli. Biochemistry 38: 7235-7242.
  • Yang L, Harroun TA, Weiss TM, Ding L, Huang HW (2001) Barrel-stave model or toroidal model? A case study on melittin pores. Biophys J 81: 1475-1785.
Document Type
Publication order reference
YADDA identifier
JavaScript is turned off in your web browser. Turn it on to take full advantage of this site, then refresh the page.