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Silver nanoparticles induce apoptosis and G2/M arrest via PKCζ-dependent signaling in A549 lung cells

  • Inorganic Compounds
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Abstract

The use of silver nanoparticles is one of the fastest growing product categories in the nanotechnology industry, with a focus on antimicrobial activity. However, thus far, toxicity data for silver nanoparticles are limited. In this study, we investigated the cytotoxic effects of silver nanoparticles (Ag NPs) and the pathway by which they affect A549 lung epithelial cells. The effects of Ag NPs on cell survival, cell cycle progression, and mRNA and protein alterations of selected cell cycle- and apoptosis-related genes were studied using formazan dye and LDH release assays, flow cytometric analysis, semi-quantitative RT-PCR, and Western blot analysis. Ag NPs reduced cell viability, increased LDH release, and modulated cell cycle distribution through the accumulation of cells at G2/M and sub-G1 phases (cell death), with a concurrent decrease in cells at G1. Ag NP treatment increased Bax and Bid mRNA levels and downregulated Bcl-2 and Bcl-w mRNAs in a dose-dependent manner. Furthermore, Ag NPs altered the mRNA levels of protein kinase C (PKC) family members. In particular, ectopic overexpression of PKCζ led to the enhancement of cellular proliferation and reduced sensitivity to Ag NPs in A549 cells. Together, these results suggest that Ag NPs induce strong toxicity and G2/M cell cycle arrest by a mechanism involving PKCζ downregulation in A549 cells.

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References

  • Ahamed M, Karns M, Goodson M, Rowe J, Hussain SM, Schlager JJ, Hong YL (2008) DNA damage response to different surface chemistry of silver nanoparticles in mammalian cells. Toxicol Appl Pharm 233:404–410

    Article  CAS  Google Scholar 

  • Alvarez-Gonzalez R, Spring H, Muller M, Burkle A (1999) Selective loss of poly(ADP-ribose) and the 85-kDa fragment of poly(ADP-ribose) polymerase in nucleoli during alkylation-induced apoptosis of HeLa cells. J Biol Chem 274:32122–32126

    Article  PubMed  CAS  Google Scholar 

  • AshaRani PV, Low Kah Mun G, Hande MP, Valiyaveettil S (2009) Cytotoxicity and genotoxicity of silver nanoparticles in human cells. ACS Nano 3:279–290

    Article  PubMed  CAS  Google Scholar 

  • Benn TM, Westerhoff P (2008) Nanoparticle silver released into water from commercially available sock fabrics (vol 42, p 4133, 2008). Environ Sci Technol 42:7025–7026

    Article  CAS  Google Scholar 

  • Cerda SR, Mustafi R, Little H, Cohen G, Khare S, Moore C, Majumder P, Bissonnette M (2006) Protein kinase C delta inhibits Caco-2 cell proliferation by selective changes in cell cycle and cell death regulators. Oncogene 25:3123–3138

    Article  PubMed  CAS  Google Scholar 

  • Charruyer A, Jean C, Colomba A, Jaffrezou JP, Quillet-Mary A, Laurent G, Bezombes C (2007) PKC zeta protects against UV-C-induced apoptosis by inhibiting acid sphingomyelinase-dependent ceramide production. Biochem J 405:77–83

    PubMed  CAS  Google Scholar 

  • Chen X, Schluesener HJ (2008) Nanosilver: a nanoproduct in medical application. Toxicol Lett 176:1–12

    Article  PubMed  CAS  Google Scholar 

  • Choi K, Kim SE, Kim JY, Yoon J, Lee JC (2008) Poly(oxyethylene)/silver nanoparticle composites as biocidal agents. J Nanosci Nanotechnol 8:5360–5362

    Article  PubMed  CAS  Google Scholar 

  • Cioffi N, Ditaranto N, Torsi L, Picca RA, De Giglio E, Sabbatini L, Novello L, Tantillo G, Bleve-Zacheo T, Zambonin PG (2005) Synthesis, analytical characterization and bioactivity of Ag and Cu nanoparticles embedded in poly-vinyl-methyl-ketone films. Anal Bioanal Chem 382:1912–1918

    Article  PubMed  CAS  Google Scholar 

  • Cohen A, Brodie C, Sarid R (2006) An essential role of ERK signalling in TPA-induced reactivation of Kaposi’s sarcoma-associated herpesvirus. J Gen Virol 87:795–802

    Google Scholar 

  • Dai TH, Tegos GP, Burkatovskaya M, Castano AP, Hamblin MR (2009) Chitosan acetate bandage as a topical antimicrobial dressing for infected burns. Antimicrob Agents Chemother 53:393–400

    Article  PubMed  CAS  Google Scholar 

  • Datta K, Li JP, Bhattacharya R, Gasparian L, Wang EF, Mukhopadhyay D (2004) Protein kinase C zeta transactivates hypoxia-inducible factor alpha by promoting its association with p300 in renal cancer. Cancer Res 64:456–462

    Article  PubMed  CAS  Google Scholar 

  • De Martino A, Filomeni G, Aquilano K, Ciriolo MR, Rotilio G (2006) Effects of water garlic extracts on cell cycle and viability of HepG2 hepatoma cells. J Nutr Biochem 17:742–749

    Article  PubMed  Google Scholar 

  • Duran N, Marcato PD, Alves OL, Souza GI, Esposito E (2005) Mechanistic aspects of biosynthesis of silver nanoparticles by several Fusarium oxysporum strains. J Nanobiotechnol 3:8

    Article  Google Scholar 

  • Fang JL, Beland FA (2009) Long-term exposure to zidovudine delays cell cycle progression, induces apoptosis, and decreases telomerase activity in human hepatocytes. Toxicol Sci 111:120–130

    Article  PubMed  CAS  Google Scholar 

  • Fukumoto S, Nishizawa Y, Hosoi M, Koyama H, Yamakawa K, Ohno S, Morii H (1997) Protein kinase C delta inhibits the proliferation of vascular smooth muscle cells by suppressing G1 cyclin expression. J Biol Chem 272:13816–13822

    Article  PubMed  CAS  Google Scholar 

  • Impellitteri CA, Tolaymat TM, Scheckel KG (2009) The speciation of silver nanoparticles in antimicrobial fabric before and after exposure to a hypochlorite/detergent solution. J Environ Qual 38:1528–1530

    Article  PubMed  CAS  Google Scholar 

  • Kanemitsu H, Yamauchi H, Komatsu M, Yamamoto S, Okazaki S, Uchida K, Nakayama H (2009) 6-Mercaptopurine (6-MP) induces cell cycle arrest and apoptosis of neural progenitor cells in the developing fetal rat brain. Neurotoxicol Teratol 31:104–109

    Article  PubMed  CAS  Google Scholar 

  • Kim SA, Kim SW, Chang S, Yoon JH, Ahn SG (2009) 5′-nitro-indirubinoxime induces G2/M cell cycle arrest and apoptosis in human KB oral carcinoma cells. Cancer Lett 274:72–77

    Article  PubMed  CAS  Google Scholar 

  • Kumar A, Vemula PK, Ajayan PM, John G (2008) Silver-nanoparticle-embedded antimicrobial paints based on vegetable oil. Nat Mater 7:236–241

    Article  PubMed  CAS  Google Scholar 

  • Kutsuzawa K, Chowdhury EH, Nagaoka M, Maruyama K, Akiyama Y, Akaike T (2006) Surface functionalization of inorganic nano-crystals with fibronectin and E-cadherin chimera synergistically accelerates trans-gene delivery into embryonic stem cells. Biochem Biophs Res Commun 350:514–520

    Article  CAS  Google Scholar 

  • Kvitek L, Soukupova J (2009) Comment on ‘Preparation and antibacterial activity of Fe3O4@ Ag nanoparticles’. Nanotechnology 20:028001

    Google Scholar 

  • Lag M, Refsnes M, Lilleaas EM, Holme JA, Becher R, Schwarze PE (2005) Role of mitogen activated protein kinases and protein kinase C in cadmium-induced apoptosis of primary epithelial lung cells. Toxicology 211:253–264

    Article  PubMed  CAS  Google Scholar 

  • Lee HY, Park HK, Lee YM, Kim K, Park SB (2007) A practical procedure for producing silver nanocoated fabric and its antibacterial evaluation for biomedical applications. Chem Commun 28:2959–2961

    Google Scholar 

  • Lee YS, Yoon HJ, Oh JH, Park HJ, Lee EH, Song CW, Yoon S (2009) 1,3-Dinitrobenzene induces apoptosis in TM4 mouse Sertoli cells: Involvement of the c-Jun N-terminal kinase (JNK) MAPK pathway. Toxicol Lett 189:145–151

    Article  PubMed  CAS  Google Scholar 

  • Liu BS, Huang TB (2008) Nanocomposites of genipin-crosslinked chitosan/silver nanoparticles—structural reinforcement and antimicrobial properties. Macromol Biosci 8:932–941

    Article  PubMed  CAS  Google Scholar 

  • Lu SY, Gao WJ, Gu HY (2008) Construction, application and biosafety of silver nanocrystalline chitosan wound dressing. Burns 34:623–628

    Article  PubMed  Google Scholar 

  • Makhluf SB, Abu-Mukh R, Rubinstein S, Breitbart H, Gedanken A (2008) Modified PVA-Fe3O4 nanoparticles as protein carriers into sperm cells. Small 4:1453–1458

    Article  PubMed  Google Scholar 

  • Morita Y, Naka T, Kawazoe Y, Fujimoto M, Nakagawa R, Fukuyama H, Nagata S, Kishimoto T (2000) Signals transducers and activators of transcription (STAT)-induced STAT inhibitor-1 (SSI-1)/suppressor of cytokine signaling-1 (SOCS-1) suppresses tumor necrosis factor α-induced cell death in fibroblasts. Proc Natl Acad Sci USA 91:5405–5410

    Google Scholar 

  • Morrow KJ Jr, Bawa R, Wei C (2007) Recent advances in basic and clinical nanomedicine. Med Clin North Am 91:805–843

    Article  PubMed  CAS  Google Scholar 

  • Mukherjee S, Dey S, Bhattacharya RK, Roy M (2009) Isothiocyanates sensitize the effect of chemotherapeutic drugs via modulation of protein kinase C and telomerase in cervical cancer cells. Mol Cell Biochem 330:9–22

    Article  PubMed  CAS  Google Scholar 

  • Muro S, Mateescu M, Gajewski C, Robinson M, Muzykantov VR, Koval M (2006) Control of intracellular trafficking of ICAM-1-targeted nanocarriers by endothelial Na+/H+ exchanger proteins. Am J Physiol Lung Can 290:L809–L817

    Article  CAS  Google Scholar 

  • Mustafi R, Cerda S, Chumsangsri A, Fichera A, Bissonnette M (2006) Protein kinase-zeta inhibits collagen I-dependent and anchorage-independent growth and enhances apoptosis of human Caco-2 cells. Mol Cancer Res 4:683–694

    Article  PubMed  CAS  Google Scholar 

  • Peng YH, Sigua CA, Karsonovich C, Murr MM (2007) Protein kinase C-zeta (PKC-zeta) regulates Kupffer cell apoptosis during experimental sepsis. J Gastrointest Surg 11:1712–1721

    Article  PubMed  Google Scholar 

  • Ron D, Kazanietz MG (1999) New insights into the regulation of protein kinase C and novel phorbol ester receptors. Faseb J 13:1658–1676

    PubMed  CAS  Google Scholar 

  • Scampicchio M, Arecchi A, Mannino S (2009) Optical nanoprobes based on gold nanoparticles for sugar sensing. Nanotechnology 20:135501

    Article  PubMed  Google Scholar 

  • Shahverdi AR, Fakhimi A, Shahverdi HR, Minaian S (2007) Synthesis and effect of silver nanoparticles on the antibacterial activity of different antibiotics against Staphylococcus aureus and Escherichia coli. Nanomed Nanotechnol 3:168–171

    Article  CAS  Google Scholar 

  • Shatos MA, Hodges RR, Oshi Y, Bair JA, Zoukhri D, Kublin C, Lashkari K, Dartt DA (2009) Role of cPKC alpha and nPKC epsilon in EGF-stimulated goblet cell proliferation. Invest Ophth Vis Sci 50:614–620

    Article  Google Scholar 

  • Sun ZM, Arendt CW, Ellmeier W, Schaeffer EM, Sunshine MJ, Gandhi L, Annes J, Petrzilka D, Kupfer A, Schwartzberg PL, Littman DR (2000) PKC-theta is required for TCR-induced NF-kappa B activation in mature but not immature T lymphocytes. Nature 404:402–407

    Article  PubMed  CAS  Google Scholar 

  • Sun RH, Gao P, Chen L, Ma DL, Wang JM, Oppenheim JJ, Zhang N (2005) Protein kinase C zeta is required for epidermal growth factor-induced chemotaxis of human breast cancer cells. Cancer Res 65:1433–1441

    Article  PubMed  CAS  Google Scholar 

  • Sung JH, Ji JH, Yoon JU, Kim DS, Song MY, Jeong J, Han BS, Han JH, Chung YH, Kim J, Kim TS, Chang HK, Lee EJ, Lee JH, Yu IJ (2008) Lung function changes in Sprague-Dawley rats after prolonged inhalation exposure to silver nanoparticles. Inhal Toxicol 20:567–574

    Article  PubMed  CAS  Google Scholar 

  • Takahashi T, Shibuya M (2001) The overexpression of PKC delta is involved in vascular endothelial growth factor-resistant apoptosis in cultured primary sinusoidal endothelial cells. Biochem Biophs Res Commun 280:415–420

    Article  CAS  Google Scholar 

  • Tulla M, Helenius J, Jokinen J, Taubenberger A, Muller DJ, Heino J (2008) TPA primes alpha 2 beta 1 integrins for cell adhesion (vol 582, p 3520, 2009). Febs Lett 582:3966

    Article  CAS  Google Scholar 

  • Venugopal J, Low S, Choon AT, Ramakrishna S (2008) Interaction of cells and nanofiber scaffolds in tissue engineering. J Biomed Mater Res B 84B:34–48

    Article  CAS  Google Scholar 

  • Vigneshwaran N, Kathe AA, Varadarajan PV, Nachane RP, Balasubramanya RH (2007) Functional finishing of cotton fabrics using silver nanoparticles. J Nanosci Nanotechnol 7:1893–1897

    Article  PubMed  CAS  Google Scholar 

  • Watkin RD, Nawrot T, Potts RJ, Hart BA (2003) Mechanisms regulating the cadmium-mediated suppression of Sp1 transcription factor activity in alveolar epithelial cells. Toxicology 184:157–178

    Article  PubMed  CAS  Google Scholar 

  • White AT, Spence FJ, Chipman JK (2005) Glutathione depletion modulates gene expression in HepG2 cells via activation of protein kinase C alpha. Toxicology 216:168–180

    Article  PubMed  CAS  Google Scholar 

  • Wong KKY, Cheung SOF, Huang LM, Niu J, Tao C, Ho CM, Che CM, Tam PKH (2009) Further evidence of the anti-inflammatory effects of silver nanoparticles. Chemmedchem 4:1129–1135

    Article  PubMed  CAS  Google Scholar 

  • Wu TT, Hsieh YH, Hsieh YS, Liu JY (2008) Reduction of PKC alpha decreases cell proliferation, migration, and invasion of human malignant hepatocellular carcinoma. J Cell Biochem 103:9–20

    Article  PubMed  CAS  Google Scholar 

  • Xin MG, Gao FQ, May WS, Flagg T, Deng XM (2007) Protein kinase C zeta abrogates the proapoptotic function of bax through phosphorylation. J Biol Chem 282:21268–21277

    Article  PubMed  CAS  Google Scholar 

  • Zhang L, Zhang J, Hu CQ, Cao J, Zhou XL, Hu YZ, He QJ, Yang B (2009) Efficient activation of p53 pathway in A549 cells exposed to L2, a novel compound targeting p53-MDM2 interaction. Anti Cancer Drug 20:416–424

    Article  CAS  Google Scholar 

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Acknowledgments

This work was supported by a 2009 General Project grant from the Ministry of Knowledge and Economy through the International Inhalation Toxicology Evaluation Technology program at the Korea Institute of Toxicology.

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Authors and Affiliations

  1. Department of Anatomy, Chungnam National University, Daejeon, 301-040, Republic of Korea

    Young Sook Lee, Dong Woon Kim & Young Ho Lee

  2. Molecular Toxicology Team, Korea Institute of Toxicology, Daejeon, 305-343, Republic of Korea

    Jung Hwa Oh, Seokjoo Yoon & Mi Sun Choi

  3. Environmental Toxicology Team, Korea Institute of Toxicology, Daejeon, 305-343, Republic of Korea

    Sung Kyu Lee & Ji Won Kim

  4. Department of Inhalation Toxicology, Korea Institute of Toxicology, Korea Research Institute of Chemical Technology, 1051, Shinjeong-dong, Jeongeup, Jeollabuk-do, 580-185, Republic of Korea

    Kyuhong Lee & Chang-Woo Song

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  1. Young Sook Lee
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Correspondence to Chang-Woo Song.

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Lee, Y.S., Kim, D.W., Lee, Y.H. et al. Silver nanoparticles induce apoptosis and G2/M arrest via PKCζ-dependent signaling in A549 lung cells. Arch Toxicol 85, 1529–1540 (2011). https://doi.org/10.1007/s00204-011-0714-1

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