Violacein induces cell death by triggering mitochondrial membrane hyperpolarization in vitro

BMC Microbiology

Lucymara Fassarella Agnez-Lima²  Tatjana Keesen de Souza Lima¹  Silvia Regina Batistuzzo de Medeiros²  Jana Dara Freires de Queiroz²  Angélica Maria de Sousa Leal² 

[1] Departamento de Biologia Celular e Molecular, Centro de Biotecnologia, Universidade Federal da Paraíba, CEP 58051-900, João Pessoa, PB, Brazil;Departamento de Biologia Celular e Genética, Universidade Federal do Rio Grande do Norte, CEP 59072-970, Natal, RN, Brazil

关键词: Mitochondrial membrane potential;    Cytotoxicity;    Oxidative stress;    Chromobacterium violaceum;    Violacein;   

Others  :  1221681
DOI  :  10.1186/s12866-015-0452-2

 received in 2014-09-30, accepted in 2015-05-27,  发布年份 2015

【 摘 要 】

Background

Violacein is a purple pigment from Chromobacterium violaceum that possesses diverse biological and pharmacological properties. Among these, pro-oxidant and antioxidant activities have been suggested. However, the cytotoxic mechanisms induced by violacein are poorly understood and the improvement in knowledge regarding these cell death mechanisms will be useful to develop new therapeutic approaches. Considering this, in our work, we investigated the pro-oxidant effects of violacein in non-tumor (CHO-K1 and MRC-5) and tumor (HeLa) cell lines, searching for a better understanding of reactive oxygen species (ROS) production and cell death induction.

Results

Cytotoxicity induced by violacein was observed in the three cell lines; however, MRC-5 and HeLa cells were shown to be more sensitive to violacein treatment. Although punctual alterations in the antioxidant apparatus and increase in oxidative stress biomarkers was observed in some violacein concentrations, no association was found between increased oxidative stress and induction of cell death. However, the increase of mitochondrial membrane potential was observed.

Conclusions

In fact, the increase of mitochondrial membrane potential in MRC-5 and HeLa cells suggests that mitochondrial membrane hyperpolarization might be the main cause of cell death triggered by violacein.

【 授权许可】

   
2015 Leal et al.

【 图 表 】

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【 参考文献 】

• [1]Subramaniam S, Ravi V, Sivasubramanian A. Synergistic antimicrobial profiling of violacein with commercial antibiotics against pathogenic micro-organisms. Pharm Biol. 2014; 52:86-90.
• [2]Cazoto LL, Martins D, Ribeiro MG, Durán N, Nakazato G. Antibacterial activity of violacein against Staphylococcus aureus isolated from bovine mastitis. J Antibiot (Tokyo). 2011; 64:395-7.
• [3]Berni E, Marcato PD, Nakazato G, Kobayashi RKT, Vacchi FI, Umbuzeiro GA, Durán N. Violacein/poly(ε-caprolactone)/chitosan nanoparticles against bovine mastitis: Antibacterial and ecotoxicity evaluation. J Phys: ConfSer. 2013; 429:012030.
• [4]Melo PS, Maria SS, Vidal BC, Haun M, Durán N. Violacein cytotoxicity and induction of apoptosis in V79 cells. In Vitro Cell Dev Biol Anim. 2000; 36:539-43.
• [5]Bromberg N, Dreyfuss JL, Regatieri CV, Palladino MV, Durán N, Nader HB, Haun M, Justo GZ. Growth inhibition and pro-apoptotic activity of violacein in Ehrlich ascites tumor. Chem Biol Interact. 2010; 186:43-52.
• [6]Queiroz KC, Milani R, Ruela-de-Sousa RR, Fuhler GM, Justo GZ, Zambuzzi WF, Duran N, Diks SH, Spek CA, Ferreira CV, Peppelenbosch MP. Violacein induces deathof resistant leukaemia cells via kinome reprogramming, endoplasmic reticulumstress and Golgi apparatus collapse. PLoS One. 2012; 7:e45362.
• [7]Leon LL, Miranda CC, De Souza AO, Durán N. Antileishmanial activity of the violacein extracted from Chromobacteriumviolaceum. J Antimicrob Chemother. 2001; 48:449-50.
• [8]May G, Brummer B, Ott H. Treatment of prophylaxis of polio and herpes virus infections comprises admin. of 3-(1,2-dihydro-5-(5-hydroxy-1H- indol-3-yl)-2-oxo-3H-pyrrole-3-ylidene)-1,3-dihydro-2H-indol-2-one. GerOffenDE 1991, 3935066.
• [9]Andrighetti-Fröhner CR, Antonio RV, Creczynski-Pasa TB, Barardi CR, Simões CM. Cytotoxicity and potential antiviral evaluation of violacein produced by Chromobacteriumviolaceum. Mem Inst Oswaldo Cruz. 2003; 98:843-8.
• [10]Durán N, Antonio RV, Haun M, Pilli RA. Biosynthesis of a trypanocide by Chromobacteriumviolaceum. World J Microbiol Biotechno. 1994; 10:686-90.
• [11]Matz C, Webb JS, Schupp PJ, Phang SY, Penesyan A, Egan S, Steinberg P, Kjelleberg S. Marine biofilm bacteria evade eukaryotic predation by targeted chemical defense. PLoS One. 2008; 3:e2744.
• [12]Lopes SC, Blanco YC, Justo GZ, Nogueira PA, Rodrigues FL, Goelnitz U, Wunderlich G, Facchini G, Brocchi M, Duran N, Costa FT. Violacein extracted from Chromobacteriumviolaceum inhibits Plasmodium growth in vitro and in vivo. Antimicrob Agents Chemother. 2009; 53:2149-52.
• [13]Antonisamy P, Ignacimuthu S. Immunomodulatory, analgesic and antipyretic effects of violacein isolated from Chromobacteriumviolaceum. Phytomedicine. 2010; 17:300-4.
• [14]Antonisamy P, Kannan P, Ignacimuthu S. Anti-diarrhoeal and ulcer-protective effects of violacein isolated from Chromobacteriumviolaceum in Wistar rats. Fundam Clin Pharmacol. 2009; 23:483-90.
• [15]Konzen M, De Marco D, Cordova CA, Vieira TO, Antônio RV, Creczynski-Pasa TB. Antioxidant properties of violacein: possible relation on its biological function. Bioorg Med Chem. 2006; 14:8307-13.
• [16]de Carvalho DD, Costa FT, Duran N, Haun M. Cytotoxic activity of violacein in human colon cancer cells. Toxicol In Vitro. 2006; 20:1514-21.
• [17]Arur S, Uche UE, Rezaul K, Fong M, Scranton V, Cowan AE, Mohler W, Han DK. Annexin I is an endogenous ligand that mediates apoptotic cell engulfment. Dev Cell. 2003; 4:587-98.
• [18]Wlodkowic D, Telford W, Skommer J, Darzynkiewicz Z. Apoptosis and beyond: cytometry in studies of programmed cell death. Methods Cell Biol. 2011; 103:55-98.
• [19]Bromberg N, Justo GZ, Haun M, Durán N, Ferreira CV. Violacein cytotoxicity on human blood lymphocytes and effect on phosphatases. J Enzyme Inhib Med Chem. 2005; 20:449-54.
• [20]Saraiva VS, Marshall JC, Cools-Lartigue J, Burnier MN. Cytotoxic effects of violacein in human uveal melanoma cell lines. Melanoma Res. 2004; 14:421-4.
• [21]Matés JM. Effects of antioxidant enzymes in the molecular control of reactive oxygen species toxicology. Toxicology. 2000; 153:83-104.
• [22]Franco-Molina MA, Mendoza-Gamboa E, Sierra-Rivera CA, Gómez-Flores RA, Zapata-Benavides P, Castillo-Tello P, Alcocer-González JM, Miranda-Hernández DF, Tamez-Guerra RS, Rodríguez-Padilla C. Antitumor activity of colloidal silver on MCF-7 human breast cancer cells. J Exp Clin Cancer Res. 2010; 29:148. BioMed Central Full Text
• [23]Mojib N, Nasti TH, Andersen DT, Attigada VR, Hoover RB, Yusuf N, Bej AK. The antiproliferative function of violacein-like purple violet pigment (PVP) from an Antarctic Janthinobacterium sp. Ant5-2 in UV-induced 2237 fibrosarcoma. Int J Dermatol. 2011; 50:1223-33.
• [24]Melo PS, de Azevedo MM, Frungillo L, Anazetti MC, Marcato PD, Durán N. Nanocytotoxicity: violacein and violacein-loaded poly (D, L-lactide-co-glycolide) nanoparticles acting on human leukemic cells. J Biomed Nanotechnol. 2009; 5:192-201.
• [25]Ferreira CV, Bos CL, Versteeg HH, Justo GZ, Durán N, Peppelenbosch MP. Molecular mechanism of violacein-mediated human leukemia cell death. Blood. 2004; 104:1459-64.
• [26]Ly JD, Grubb DR, Lawen A. The mitochondrial membrane potential (deltapsi(m))in apoptosis; an update. Apoptosis. 2003; 8:115-28.
• [27]Perl A, Gergely P, Nagy G, Koncz A, Banki K. Mitochondrial hyperpolarization: a checkpoint of T-cell life, death and autoimmunity. Trends Immunol. 2004; 25:360-7.
• [28]Banki K, Hutter E, Gonchoroff NJ, Perl A. Elevation of mitochondrial transmembrane potential and reactive oxygen intermediate levels are early events and occur independently from activation of caspases in Fassignaling. J Immunol. 1999; 162(3):1466-79.
• [29]VandenBerghe T, Vanlangenakker N, Parthoens E, Deckers W, Devos M, Festjens N, Guerin CJ, Brunk UT, Declercq W, Vandenabeele P. Necroptosis, necrosis and secondary necrosis converge on similar cellular disintegration features. Cell Death Differ. 2010; 6:922-30.
• [30]Rettori D, Duran N. Production, extraction and purificationof violacein: an antibiotic pigment producedby Chromobacteriumviolaceum. World J Microbiol Biotechnol. 1998; 14:685-8.
• [31]Kaltenbach JP, Kaltenbach MH, Lyons WB. Nigrosin as a dye for differentiating live and dead ascites cells. Exp Cell Res. 1958; 15:112-7.
• [32]Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976; 72:248-54.
• [33]Petta TB, Nakajima S, Zlatanou A, Despras E, Couve-Privat S, Ishchenko A, Sarasin A, Yasui A, Kannouche P. Human DNA polymerase iota protects cells against oxidative stress. EMBO J. 2008; 27:2883-95.
• [34]Hawkins CL, Morgan PE, Davies MJ. Quantification of protein modification by oxidants. Free Radic Biol Med. 2009; 46:965-88.
• [35]Li JJ, Tang Q, Li Y, Hu BR, Ming ZY, Fu Q, Qian JQ, Xiang JZ. Role of oxidative stress in the apoptosis of hepatocellular carcinoma induced by combination of arsenic trioxide and ascorbic acid. Acta Pharmacol Sin. 2006; 27:1078-84.