Cytotoxic effect of pyocyanin on human pancreatic cancer cell line (Panc-1)

Document Type : Original Article


Department of Microbiology, Tehran North Branch, Islamic Azad University, Tehran, Iran


Objective(s): Pyocyanin, a blue-green pigment produced by Pseudomonas aeruginosa, interferes with host redox cycles, which can lead to generation of reactive oxygen species and progressive cellular oxidative damage. The aim of this study was to assess the influence of pyocyanin on human pancreatic cancer cell line.
Materials and Methods: Polymerase Chain Reaction (PCR) was applied to confirm the existence of a specific pyocyanin producing gene (phzM). The pigment was then characterized by UV-visible, FT-IR, and HPLC analysis. Panc-1 cells were treated by different concentrations of pyocyanin and their cytotoxic effect as well as the induction of apoptosis/necrosis were evaluated by XTT assay and        flow cytometry.
Results: An overnight pyocyanin treatment resulted in significant cell reduction in a concentration-dependent manner. Inhibition rate of 6 pyocyanin (the highest concentration) extracted from clinical and soil isolates of P. aeruginosa were 98.69±0.23 and 89.88±1.86%, respectively, which decreased as the pyocyanin concentration lessened. Pyocyanin could also induce dose-dependent apoptosis/necrosis in Panc-1 cells after 24 hr.
Conclusion: We reported, for the first time, cytotoxic effects of pyocyanin against human pancreatic cancer cell line. Considering this effect of the pigment, study on pyocyanin as a potential anti-tumor biodrug requires further studies.


Main Subjects

1. Lau GW, Hassett DJ, Ran H, Kong F. The role of pyocyanin in Pseudomonas aeruginosa infection. Trends Mol Med 2004; 10:599-606.
2. Muller M, Li Z, Maitz PK. Pseudomonas pyocyanin inhibits wound repair by inducing premature cellular senescence: role for p38 mitogen-activated protein kinase. Burns 2009; 35:500-508.
3. Budzikiewicz H. Secondary metabolites from fluorescent pseudomonads. FEMS Microbiol Rev 1993; 104:209-228.
4. Hassett D, Charniga L, Bean K, Ohman D, Cohen M. Response of Pseudomonas aeruginosa to pyocyanin: mechanisms of resistance, antioxidant defenses, and demonstration of a manganese-cofactored superoxide dismutase. Infect Immun 1992; 60:328-336.
5. Denning GM, Railsback MA, Rasmussen GT, Cox CD, Britigan BE. Pseudomonas pyocyanine alters calcium signaling in human airway epithelial cells. Am J Physiol: Lung Cell Mol Physiol 1998; 274:L893-L900.
6. Usher LR, Lawson RA, Geary I, Taylor CJ, Bingle CD, Taylor GW, et al. Induction of neutrophil apoptosis by the Pseudomonas aeruginosa exotoxin pyocyanin: a potential mechanism of persistent infection. J Immunol 2002; 168:1861-1868.
7. Laursen JB, Nielsen J. Phenazine natural products: biosynthesis, synthetic analogues, and biological activity. Chem Rev 2004; 104:1663-1686.
8. Mavrodi DV, Blankenfeldt W, Thomashow LS. Phenazine compounds in fluorescent Pseudomonas spp. biosynthesis and regulation. Annu Rev Phytopathol 2006; 44:417-445.
9. Zhao J, Wu Y, Alfred A, Wei P, Yang S. Anticancer effects of pyocyanin on HepG2 human hepatoma cells. Lett Appl Microbiol 2014; 58:541-548.
10. Hassani HH, Hasan HM, Al-Saadi A, Ali AM, Muhammad MH. A comparative study on cytotoxicity and apoptotic activity of pyocyanin produced by wild type and mutant strains of Pseudomonas aeruginosa. Eur J Exp Biol 2012; 2:1389-1394.
11. Lozano R, Naghavi M, Foreman K, Lim S, Shibuya K, Aboyans V, et al. Global and regional mortality from 235 causes of death for 20 age groups in 1990 and 2010: a systematic analysis for the Global Burden of Disease Study 2010. The Lancet 2013; 380:2095-2128.
12. Al Haddad AH, Adrian TE. Challenges and future directions in therapeutics for pancreatic ductal adenocarcinoma. Expert Opin Invest Drugs 2014; 23:1499-1515.
13. Nakaike S, Yamagishi T, Nanaumi K, Otomo S, Tsukagoshi S. Cell‐killing activity and kinetic analysis of a novel antitumor compound NC‐190, a benzo[a]phenazine derivative. Jpn J Cancer Res 1992; 83:402-409.
14. Moayedi A, Nowroozi J, Akhavan Sepahy A. Effect of fetal and adult bovine serum on pyocyanin production in Pseudomonas aeruginosa isolated from clinical and soil samples. Iran J Basic Med Sci 2017; 20:1331-1338.
15. Gohain N, Thomashow LS, Mavrodi DV, Blankenfeldt W. The purification, crystallization and preliminary structural characterization of PhzM, a phenazine-modifying methyltransferase from Pseudomonas aeruginosa. Acta Crystallogr Sect F Struct Biol Cryst Commun 2006; 62:887-890.
16. Jayaseelan S, Ramaswamy D, Dharmaraj S. Pyocyanin: production, applications, challenges and new insights. World J Microbiol Biotechnol 2014; 30:1159-1168.
17. Ra’oof W, Latif I. In vitro study of the swarming phenomena and antimicrobial activity of pyocyanin produced by Pseudomonas aeruginosa isolated from different human infections. Eur J Sci Res 2010; 47:405-421.
18. Mavrodi DV, Bonsall RF, Delaney SM, Soule MJ, Phillips G, Thomashow LS. Functional analysis of genes for biosynthesis of pyocyanin and phenazine-1-carboxamide from Pseudomonas aeruginosa PAO1. J Bacteriol 2001; 183:6454-6465.
19. Ohfuji K, Sato N, Hamada-Sato N, Kobayashi T, Imada C, Okuma H, et al. Construction of a glucose sensor based on a screen-printed electrode and a novel mediator pyocyanin from Pseudomonas aeruginosa. Biosens Bioelectron 2004; 19:1237-1244.
20. El-Fouly M, Sharaf A, Shahin A, El-Bialy HA, Omara A. Biosynthesis of pyocyanin pigment by Pseudomonas aeruginosa. J Radiat Res Appl Sci 2015; 8:36-48.
21. Mohammed HA, Yossef HS, Mohammad FI. The cytotoxicity effect of pyocyanin on human hepatocellular carcinoma cell line (HepG2). Iraqi J Sci 2014; 55:668-674.
22. O’Malley YQ, Reszka KJ, Spitz DR, Denning GM, Britigan BE. Pseudomonas aeruginosa pyocyanin directly oxidizes glutathione and decreases its levels in airway epithelial cells. Am J Physiol: Lung Cell Mol Physiol 2004; 287:L94-L103.
23. Dickinson DA, Forman HJ. Cellular glutathione and thiols metabolism. Biochem Pharmacol 2002; 64:1019-1026.
24. Jacob C, Jamier V, Ba LA. Redox active secondary metabolites. Curr Opin Chem Biol 2011; 15:149-155.
25. Wilson R, Sykes D, Watson D, Rutman A, Taylor G, Cole P. Measurement of Pseudomonas aeruginosa phenazine pigments in sputum and assessment of their contribution to sputum sol toxicity for respiratory epithelium. Infect Immun 1988; 56:2515-2517.