Abstract
Metal based drugs have successfully been used in both the detection and treatment of different disease states. The antibacterial features of metal ion silver are well documented. Most recently, metal ion silver has been tested and applied in anticancer activity. The present study observed the cytotoxic, anti-proliferative and apoptotic effects of metal complex silver nitrate in H-ras transformed 5RP7 cell lines for 24 h. In addition, the toxic effects of silver nitrate was investigated on NIH/3T3 primary mouse embryonic fibroblast cells for 24 h. Cytotoxic effects were determined by MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) assay. Apoptosis and necrosis were evaluated by flow cytometric analysis (Annexin-V FITC/PI). Caspase-3 activation was researched by flow cytometric analysis. Apoptotic morphology was observed by DAPI staining. Structure and ultra-structure changes of cells were assessed using transmission electron microscopy. The results indicate silver nitrate has high cytotoxicity and a strong capacity to induce apoptosis in H-ras 5RP7 cells. Furthermore silver nitrate was not toxic against NIH/3T3 primary mouse embryonic fibroblast cells at low doses for 24 h.





Similar content being viewed by others
Abbreviations
- DMEM:
-
Dulbecco’s modified Eagle medium
- FBS:
-
Fetal bovine serum
- PBS:
-
Phosphate buffer solution
- MTT:
-
(3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide)
- DAPI:
-
Diamidino-2-pheylindole
- PI:
-
Propidium iodid
- FITC:
-
Fluorescein isothiocyanate
- DMSO:
-
Dimethylsulfoxide
- TEM:
-
Transmission electron microscopy
References
Ahmad S, Isab AA, Ali S, Al-Arfaj AR (2006) Perspectives in bioinorganic chemistry of some metal based therapeutic agents. Polyhedron 25:1633–1645
Akalin Ciftci G, Ulusoylar Yıldırım S, Altıntop MD, Kaplancıklı ZA (2014) Induction of apoptosis in lung adenocarcinoma and glioma cells by some oxadiazole derivatives. Med Chem Res 23:3353–3362
Alnemri ES, Livingston DJ, Nicholson DW, Salvesen G, Thornberry NA, Wong WW, Yuan J (1996) Human ICE/CED-3 protease nomenclature. Cell 87:171
Arora S, Jain J, Rajwade JM, Paknikar KM (2008) Cellular responses induced by silver nanoparticles: in vitro studies. Toxicol Lett 179:93–100
Berners-Price SJ, Bowen RJ, Galettis P, Healy PC, Mckeage MJ (1999) Structural and solution chemistry of gold(I) and silver(I) complexes of bidentate pyridyl phosphines: selective antitumour agents. Coord Chem Rev 185–186:823–836
Boca SC, Potara M, Gabudean AM, Juhem A, Baldeck PL, Astilean S (2011) Chitosan-coated triangular silver nanoparticles as a novel class of biocompatible, highly effective photothermal transducers for in vitro cancer cell therapy. Cancer Lett 311:131–140
Burghardt RC, Droleskey R (2006) Transmission electron microscopy. Curr Protoc Microbiol 3:2B.1.1–2B.1.39
Cavicchioli M, Massabni AC, Heinrich TA, Costa-Neto CM, Abrao EP, Fonseca BAL, Castellano EE, Corbi PP, Lustri WR, Leite CQF (2010) Pt(II) and Ag(I) complexes with acesulfame: crystal structure and a study of their antitumoral, antimicrobial and antiviral activities. J Inorg Biochem 104:533–540
Dinda J, Nandy A, Rana BK, Bertolasi V, Saha KD, Bielawski CW (2014) Cytotoxicity of silver(I), gold(I) and gold(III) complexes of a pyridine wingtip substituted annelated N-heterocyclic carbene. RSC Adv 4:60776–60784
Foldbjerg R, Olesena P, Hougaard M, Danga DA, Hoffmannc HJ, Autrupa H (2009) PVP-coated silver nanoparticles and silver ions induce reactive oxygen species, apoptosis and necrosis in THP-1 monocytes. Toxicol Lett 190:156–162
Frazer RA (2012) Use of silver nanoparticles in HIV treatment protocols: a research proposal. J Nanomed Nanotechnol 3:127
Gopinath P, Gogoi SK, Sanpui P, Paul A, Chattopadhyay A, Ghosh SS (2010) Signaling gene cascade in silver nanoparticle induced apoptosis. Colloids Surf B 77:240–245
Greulich C, Diendorf J, Geßmann J, Simon T, Habijan T, Eggeler G, Schildhauer TA, Epple M, Köller M (2011) Cell type-specific responses of peripheral blood mononuclear cells to silver nanoparticles. Acta Biomater 7:3505–3514
Human Z, Munyaneza A, Omondi B, Sanabria NM, Meijboom R, Cronje MJ (2015) The induction of cell death by phosphine silver(I) thiocyanate complexes in SNO-esophageal cancer cells. Biometals 28:219–228
Kaba SI, Egorova EM (2015) In vitro studies of the toxic effects of silver nanoparticles on HeLa and U937 cells. Nanotechnol Sci Appl 8:19–29
Koch M, Kiefer S, Cavelius C, Kraegeloh A (2012) Use of a silver ion selective electrode to assess mechanisms responsible for biological effects of silver nanoparticles. J Nanopart Res 14:646
Kolesarova A, Capcarova M, Sirotkin AV, Medvedova M, Kovacik J (2011) In vitro assessment of silver effect on porcine ovarian granulosa cells. J Trace Elem Med Bio 25:166–170
Krishnaraj C, Muthukumaran P, Ramachandran R, Balakumaran MD, Kalaichelvan PT (2014) Acalypha indica Linn: Biogenic synthesis of silver and gold nanoparticles and their cytotoxic effects against MDA-MB-231, human breast cancer cells. Biotechnol Rep 4:42–49
Liu J, Sonshine DA, Shervani S, Hurt RH (2010) Controlled release of biologically active silver from nanosilver surfaces. ACS Nano 4:6903–6913
Medvetz DA, Hindi KM, Panzner MJ, Ditto AJ, Yun YH, Youngs WJ (2008) Anticancer activity of Ag(I) N-heterocyclic carbene complexes derived from 4,5-dichloro-1H-imidazole. Met Based Drugs 2008:384010
Miao A-J, Luo Z, Chen C-S, Chin W-C, Santschi PH, Quigg A (2010) Intracellular uptake: a possible mechanism for silver engineered nanoparticle toxicity to a freshwater alga Ochromonas danica. PLoS ONE 12:e15196
Miura N, Shinohara Y (2009) Cytotoxic effect and apoptosis induction by silver nanoparticles in HeLa cells. Biochem Biophys Res Commun 390:733–737
Mossmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65:55–63
Müller G, Kramer A (2008) Biocompatibility index of antiseptic agents by parallel assessment of antimicrobial activity and cellular cytotoxicity. J Antimicrob Chemother 61:1281–1287
Nagy G, Turani M, Kovacs KE, Banfalvi G (2011) Chromatin changes upon silver nitrate treatment in human keratinocyte HaCaT and K562 erythroleukemia cells. In: Banfalvi G (ed) Cellular effects of heavy metals. Springer, Berlin, pp 195–217
Navarro E, Piccapietra F, Wagner B, Marconi F, Kaegi R, Odzak N, Sigg L, Behra R (2008) Toxicity of silver nanoparticles to Chlamydomonas reinhardtii. Environ Sci Technol 42:8959–8964
Perry DK, Smyth MJ, Stennicke HR, Salvesen GS, Duriez P, Poirier GG et al (1997) Zinc is a potent inhibitor of the apoptotic protease, caspase-3. A novel target for zinc in the inhibition of apoptosis. J Biol Chem 272:18530–18533
Sainz RM, Mayo JC, Rodriguez C, Tan DX, Lopez-Burillo S, Reiter RJ (2003) Melatonin and cell death: differential actions on apoptosis in normal and cancer cells. Cell Mol Life Sci 60:1407–1426
Siciliano TJ, Deblock MC, Hindi KM, Durmus S, Panzner MJ, Tessier CA, Youngs WJ (2011) Synthesis and anticancer properties of gold(I) and silver(I) N-heterocyclic carbene complexes. J Organomet Chem 696:1066–1071
Sotiriou GA, Pratsinis SE (2010) Antibacterial activity of nanosilver ions and particles. Environ Sci Technol 44:5649–5654
Tarnowski BI, Spinale FG, Nicholson JH (1991) DAPI as a useful stain for nuclear quantitation. Biotechnol Histochem 66:297–302
Teyssot M-L, Jarrousse A-S, Manin M, Chevry A, Roche S, Norre F, Beaudoin C, Morel L, Boyer D, Mahiou R, Gautier A (2009) Metal-NHC complexes: a survey of anti-cancer properties. Dalton Trans 35:6894–6902
Thati B, Noble A, Creaven BS, Walsh M, McCann M, Devereux M, Kavanagh K, Egan DA (2009) Role of cell cycle events and apoptosis in mediating the anti-cancer activity of a silver(I) complex of 4-hydroxy-3-nitro-coumarin-bis(phenanthroline) in human malignant cancer cells. Eur J Pharmacol 602:203–214
U.S. Environmental Protection Agency (1992) Integrated risk information system (IRIS). Environmental Criteria and Assessment Office of Environmental Assessment, Cincinnati
Vermes I, Haanen C, Steffens-Nakken H, Reutelingsperger C (1995) A novel assay for apoptosis. Flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled Annexin V. J Immunol Methods 184:39–51
Yamazaki T, Yamazaki A, Hibino Y, Chowdhury SA, Yokote Y, Kanda Y, Kunii S, Sakagami H, Nakajima H, Shimada J (2006) Biological impact of contact with metals on cells. In Vivo 20:605–611
Acknowledgments
This article was presented as a poster at The American Society for Cell Biology Annual Meeting December 15–19, 2012, San Francisco, CA.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Kaplan, A., Akalin Ciftci, G. & Kutlu, H.M. Cytotoxic, anti-proliferative and apoptotic effects of silver nitrate against H-ras transformed 5RP7. Cytotechnology 68, 1727–1735 (2016). https://doi.org/10.1007/s10616-015-9922-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10616-015-9922-5