Advertisement

The Nucleus

pp 1–12 | Cite as

Cytotoxic effect of green synthesized silver nanoparticles in MCF7 and MDA-MB-231 human breast cancer cells in vitro

  • Arindam Bandyopadhyay
  • Bishnupada Roy
  • Pallab Shaw
  • Paritosh Mondal
  • Maloy Kr. Mondal
  • Pranesh Chowdhury
  • Shelley Bhattacharya
  • Ansuman ChattopadhyayEmail author
Original Article
  • 22 Downloads

Abstract

With the incessant rise in the cancer burden worldwide it is a dire need to develop anticancer agents that will offer negligible or no side effects and at the same time will be economically feasible. In this study, we utilized the principle of green chemistry where tyrosine and chitosan were used as reducer and stabilizer respectively to synthesize biocompatible silver nanoparticles. They were characterized by ultraviolet–visible spectroscopy, transmission electron microscopy and dynamic light scattering technique and found to be spherical with average diameter of 13–22 nm. Their toxicity was evaluated in MCF7 and MDA-MB-231 human breast cancer cell lines. MTT assay revealed excellent cytotoxic effect with IC50 values as low as 6.4 and 6.56 ppb respectively after 48 h of treatment. Intriguingly, they showed minimum toxicity in normal human peripheral blood lymphocytes at these effective concentrations. Cytomorphological alteration, ROS generation (DCFDA analysis) and nuclear fragmentation (Hoechst staining) were pronounced in both cancer cell lines following treatment. These nanoparticles also promoted expression and nuclear translocation of Nrf2 as an antioxidant response which was revealed by Western blot and immunofluorescence studies respectively. ‘Apoptosis assay’ confirmed the presence of apoptosis and ‘Caspase-8 activity assay’ revealed absence of the extrinsic apoptosis pathway. Western blot data (upregulation of p21, Bax/Bcl2 ratio, Caspase-9, Caspase-3 and cleaved PARP1) established the occurrence of intrinsic apoptosis pathway following cell cycle arrest. To conclude, the green synthesized silver nanoparticles are cytotoxic to cancer cells and can be considered as effective and safe cytotoxic agents in breast cancer therapeutics.

Keywords

Silver nanoparticles Chitosan Breast cancer Intrinsic apoptosis 

Notes

Acknowledgements

The authors express their gratitude to DBT (Grant No. BT/473/NE/TBP/2013 dated 13.02.2014), India and CSIR (Award No. 09/202(0057)/2016-EMR-I dated 20.10.2016), India for their financial assistance. AB and PM are grateful to CSIR for their fellowships. Meritorious Fellowship from UGC, India is gratefully acknowledged by PS.

Compliance with ethical standards

Conflict of interest

The authors declare that there is no conflict of interest.

References

  1. 1.
    Al-Otaibi WA, Alkhatib MH, Wali AN. Cytotoxicity and apoptosis enhancement in breast and cervical cancer cells upon coadministration of mitomycin C and essential oils in nanoemulsion formulations. Biomed Pharmacother. 2018;106:946–55.PubMedCrossRefGoogle Scholar
  2. 2.
    Amaral I, Silva C, Correia-Branco A, Martel F. Effect of metformin on estrogen and progesterone receptor-positive (MCF-7) and triple-negative (MDA-MB-231) breast cancer cells. Biomed Pharmacother. 2018;102:94–101.PubMedCrossRefGoogle Scholar
  3. 3.
    Azizi M, Ghourchian H, Yazdian F, Bagherifam S, Bekhradnia S, Nyström B. Anti-cancerous effect of albumin coated silver nanoparticles on MDA-MB 231 human breast cancer cell line. Sci Rep. 2017;7:1–18.CrossRefGoogle Scholar
  4. 4.
    Bandyopadhyay A, Banerjee PP, Shaw P, Mondal MK, Das VK, Chowdhury P, et al. Cytotoxic and mutagenic effects of Thuja occidentalis mediated silver nanoparticles on human peripheral blood lymphocytes. Mater Focus. 2017;6:290–6.CrossRefGoogle Scholar
  5. 5.
    Banerjee PP, Bandyopadhyay A, Harsha SN, Policegoudra RS, Bhattacharya S, Karak N, et al. Mentha arvensis (Linn.)-mediated green silver nanoparticles trigger caspase 9-dependent cell death in MCF7 and MDA-MB-231 cells. Breast Cancer Targets Ther. 2017;9:265–78.CrossRefGoogle Scholar
  6. 6.
    Barua S, Banerjee PP, Sadhu A, Sengupta A, Chatterjee S, Sarkar S, et al. Silver nanoparticles as antibacterial and anticancer materials against human breast, cervical and oral cancer cells. J Nanosci Nanotechnol. 2017;17:968–76.PubMedCrossRefGoogle Scholar
  7. 7.
    Bhattacharyya SS, Das J, Das S, Samadder A, Das D, De A, et al. Rapid green synthesis of silver nanoparticles from silver nitrate by a homeopathic mother tincture Phytolacca Decandra. Zhong Xi Yi Jie He Xue Bao. 2012;10:546–54.PubMedCrossRefGoogle Scholar
  8. 8.
    Bhola PD, Letai A. Mitochondria—judges and executioners of cell death sentences. Mol Cell. 2016;61:695–704.PubMedPubMedCentralCrossRefGoogle Scholar
  9. 9.
    Bøyum A. Isolation of lymphocytes, granulocytes and macrophages. Scand J Immunol. 1976;5:9–15.PubMedCrossRefGoogle Scholar
  10. 10.
    Castro-Aceituno V, Ahn S, Simu SY, Singh P, Mathiyalagan R, Lee HA, et al. Anticancer activity of silver nanoparticles from Panax ginseng fresh leaves in human cancer cells. Biomed Pharmacother. 2016;84:158–65.PubMedCrossRefGoogle Scholar
  11. 11.
    Chaudhuri AR, Nussenzweig A. The multifaceted roles of PARP1 in DNA repair and chromatin remodelling. Nat Rev Mol Cell Biol. 2017;18:610–21.CrossRefGoogle Scholar
  12. 12.
    Chen MC, Mi FL, Liao ZX, Hsiao CW, Sonaje K, Chung MF, et al. Recent advances in chitosan-based nanoparticles for oral delivery of macromolecules. Adv Drug Deliv Rev. 2013;65:865–79.PubMedCrossRefGoogle Scholar
  13. 13.
    Comşa Ş, Cîmpean AM, Raica M. The story of MCF-7 breast cancer cell line: 40 years of experience in research. Anticancer Res. 2015;35:3147–54.PubMedGoogle Scholar
  14. 14.
    Das S, Das J, Samadder A, Bhattacharyya SS, Das D, Khuda-Bukhsh AR. Biosynthesized silver nanoparticles by ethanolic extracts of Phytolacca decandra, Gelsemium sempervirens, Hydrastis canadensis and Thuja occidentalis induce differential cytotoxicity through G2/M arrest in A375 cells. Colloids Surf B Biointerfaces. 2013;101:325–36.PubMedCrossRefGoogle Scholar
  15. 15.
    Das S, Khuda-Bukhsh AR. PLGA-loaded nanomedicines in melanoma treatment: future prospect for efficient drug delivery. Indian J Med Res. 2016;144:181–93.PubMedPubMedCentralCrossRefGoogle Scholar
  16. 16.
    De Matteis V, Cascione M, Toma C, Leporatti S. Silver nanoparticles: synthetic routes, in vitro toxicity and theranostic applications for cancer disease. Nanomaterials. 2018;8:1–23.CrossRefGoogle Scholar
  17. 17.
    Elinav E, Peer D. Harnessing nanomedicine for mucosal theranostics—a silver bullet at last? ACS Nano. 2013;7:2883–90.PubMedCrossRefGoogle Scholar
  18. 18.
    Escoll M, Gargini R, Cuadrado A, Anton IM, Wandosell F. Mutant p53 oncogenic functions in cancer stem cells are regulated by WIP through YAP/TAZ. Oncogene. 2017;36:3515–27.PubMedCrossRefGoogle Scholar
  19. 19.
    Franci G, Falanga A, Galdiero S, Palomba L, Rai M, Morelli G, et al. Silver nanoparticles as potential antibacterial agents. Molecules. 2015;20:8856–74.PubMedPubMedCentralCrossRefGoogle Scholar
  20. 20.
    Fulda S, Debatin KM. Extrinsic versus intrinsic apoptosis pathways in anticancer chemotherapy. Oncogene. 2006;25:4798–811.PubMedCrossRefGoogle Scholar
  21. 21.
    Georgakilas AG, Martin OA, Bonner WM. p21: a two-faced genome guardian. Trends Mol Med. 2017;23:310–9.PubMedCrossRefGoogle Scholar
  22. 22.
    Huang X, Qiao Y, Zhou Y, Ruan Z, Kong Y, Li G, et al. Ureaplasma spp. lipid-associated membrane proteins induce human monocyte U937 cell cycle arrest through p53-independent p21 pathway. Int J Med Microbiol. 2018;308:819–28.PubMedCrossRefGoogle Scholar
  23. 23.
    Ingallina E, Sorrentino G, Bertolio R, Lisek K, Zannini A, Azzolin L, et al. Mechanical cues control mutant p53 stability through a mevalonate–RhoA axis. Nat Cell Biol. 2018;20:28–35.PubMedCrossRefGoogle Scholar
  24. 24.
    Jin R, Cao YC, Hao E, Métraux GS, Schatz GC, Mirkin CA. Controlling anisotropic nanoparticle growth through plasmon excitation. Nature. 2003;425:487–90.PubMedCrossRefGoogle Scholar
  25. 25.
    Karimian A, Ahmadi Y, Yousefi B. Multiple functions of p21 in cell cycle, apoptosis and transcriptional regulation after DNA damage. DNA Repair. 2016;42:63–71.PubMedCrossRefGoogle Scholar
  26. 26.
    Kaufmann SH, Desnoyers S, Ottaviano Y, Davidson NE, Poirier GG. Specific proteolytic cleavage of poly(ADP-ribose) polymerase: an early marker of chemotherapy-induced apoptosis. Cancer Res. 1993;53:3976–85.PubMedGoogle Scholar
  27. 27.
    Khan Z, Singh T, Hussain JI, Obaid AY, Al-Thabaiti SA, El-Mossalamy EH. Starch-directed green synthesis, characterization and morphology of silver nanoparticles. Colloids Surf B Biointerfaces. 2013;102:578–84.PubMedCrossRefGoogle Scholar
  28. 28.
    Kroll A, Pillukat MH, Hahn D, Schnekenburger J. Interference of engineered nanoparticles with in vitro toxicity assays. Arch Toxicol. 2012;86:1123–36.PubMedCrossRefGoogle Scholar
  29. 29.
    Liang Y, Yan C, Schor NF. Apoptosis in the absence of caspase 3. Oncogene. 2001;20:6570–8.PubMedCrossRefGoogle Scholar
  30. 30.
    Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951;193:265–75.PubMedGoogle Scholar
  31. 31.
    Ma Q. Role of Nrf2 in Oxidative Stress and Toxicity. Annu Rev Pharmacol Toxicol. 2013;53:401–26.PubMedPubMedCentralCrossRefGoogle Scholar
  32. 32.
    Mao BH, Chen ZY, Wang YJ, Yan SJ. Silver nanoparticles have lethal and sublethal adverse effects on development and longevity by inducing ROS-mediated stress responses. Sci Rep. 2018;8:1–16.CrossRefGoogle Scholar
  33. 33.
    Mittal S, Pandey AK. Cerium oxide nanoparticles induced toxicity in human lung cells: role of ROS mediated DNA damage and apoptosis. Biomed Res Int. 2014;2014:1–14.CrossRefGoogle Scholar
  34. 34.
    Mohammed M, Syeda J, Wasan K, Wasan E. An overview of chitosan nanoparticles and its application in non-parenteral drug delivery. Pharmaceutics. 2017;9:1–26.CrossRefGoogle Scholar
  35. 35.
    Mukherjee S, Mitra I, Fouzder C, Mukherjee S, Ghosh S, Chatterji U, et al. Effect of Pt(II) complexes on cancer and normal cells compared to clinically used anticancer drugs: cell cycle analysis, apoptosis and DNA/BSA binding study. J Mol Liq. 2017;247:126–40.CrossRefGoogle Scholar
  36. 36.
    Nguyen T, Nioi P, Pickett CB. The Nrf2-antioxidant response element signaling pathway and its activation by oxidative stress. J Biol Chem. 2009;284:13291–5.PubMedPubMedCentralCrossRefGoogle Scholar
  37. 37.
    Pan Y, Ye C, Tian Q, Yan S, Zeng X, Xiao C, et al. miR-145 suppresses the proliferation, invasion and migration of NSCLC cells by regulating the BAX/BCL-2 ratio and the caspase-3 cascade. Oncol Lett. 2018;15:4337–43.PubMedPubMedCentralGoogle Scholar
  38. 38.
    Pascal JM. The comings and goings of PARP-1 in response to DNA damage. DNA Repair. 2018;71:177–82.PubMedPubMedCentralCrossRefGoogle Scholar
  39. 39.
    Reyes J, Chen JY, Stewart-Ornstein J, Karhohs KW, Mock CS, Lahav G. Fluctuations in p53 signaling allow escape from cell-cycle arrest. Mol Cell. 2018;71:581–91.PubMedPubMedCentralCrossRefGoogle Scholar
  40. 40.
    Romanov VS, Rudolph KL. p21 shapes cancer evolution. Nat Cell Biol. 2016;18:722–4.PubMedCrossRefGoogle Scholar
  41. 41.
    Roy B, Mukherjee S, Mukherjee N, Chowdhury P, Babu SP. Design and green synthesis of polymer inspired nanoparticles for the evaluation of their antimicrobial and antifilarial efficiency. RSC Adv. 2014;4:34487–99.CrossRefGoogle Scholar
  42. 42.
    Satchell P, Gutmann J, Witherspoon D. Apoptosis: an introduction for the endodontist. Int Endod J. 2003;36:237–45.PubMedCrossRefGoogle Scholar
  43. 43.
    Shalini S, Dorstyn L, Dawar S, Kumar S. Old, new and emerging functions of caspases. Cell Death Differ. 2015;22:526–39.PubMedCrossRefGoogle Scholar
  44. 44.
    Wang H, Zhang G. Endoplasmic reticulum stress-mediated autophagy protects against β, β-dimethylacrylshikonin-induced apoptosis in lung adenocarcinoma cells. Cancer Sci. 2018;109:1889–901.PubMedPubMedCentralCrossRefGoogle Scholar
  45. 45.
    Yuan YG, Zhang S, Hwang JY, Kong IK. Silver nanoparticles potentiates cytotoxicity and apoptotic potential of camptothecin in human cervical cancer cells. Oxid Med Cell Longev. 2018;2018:1–21.Google Scholar
  46. 46.
    Zhang XF, Liu ZG, Shen W, Gurunathan S. Silver nanoparticles: synthesis, characterization, properties, applications, and therapeutic approaches. Int J Mol Sci. 2016;17:1–34.Google Scholar
  47. 47.
    Zhu L, Han MB, Gao Y, Wang H, Dai L, Wen Y, et al. Curcumin triggers apoptosis via upregulation of Bax/Bcl-2 ratio and caspase activation in SW872 human adipocytes. Mol Med Rep. 2015;12:1151–6.PubMedCrossRefGoogle Scholar

Copyright information

© Archana Sharma Foundation of Calcutta 2019

Authors and Affiliations

  1. 1.Department of ZoologyVisva-BharatiSantiniketanIndia
  2. 2.Department of ChemistryVisva-BharatiSantiniketanIndia

Personalised recommendations