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. 2021 Jan 20;11(2):256.
doi: 10.3390/nano11020256.

Starch-Capped AgNPs' as Potential Cytotoxic Agents against Prostate Cancer Cells

Mariana Morais  1   2   3 , Vera Machado  1 , Francisca Dias  1 , Carlos Palmeira  4   5   6 , Gabriela Martins  4   5 , Magda Fonseca  7 , Catarina S M Martins  7 , Ana Luísa Teixeira  1 , João A V Prior  7 , Rui Medeiros  1   2   3   6   8
Affiliations

Starch-Capped AgNPs' as Potential Cytotoxic Agents against Prostate Cancer Cells

Mariana Morais et al. Nanomaterials (Basel). .

Abstract

One of the major therapeutic approaches of prostate cancer (PC) is androgen deprivation therapy (ADT), but patients develop resistance within 2-3 years, making the development of new therapeutic approaches of great importance. Silver nanoparticles (AgNPs) synthesized through green approaches have been studied as anticancer agents because of their physical-chemical properties. This study explored the cytotoxic capacity of starch-capped AgNPs, synthesized through green methods, in LNCaP and in PC-3 cells, a hormonal-sensitive and hormone-resistant PC cell line, respectively. These AgNPs were synthesized in a microwave pressurized synthesizer and characterized by ultraviolet-visible (UV-Vis) spectroscopy, transmission electron microscopy (TEM), and energy-dispersive X-ray spectroscopy (EDX). Their cytotoxicity was assessed regarding their ability to alter morphological aspect (optical microscopy), induce damage in cytoplasmic membrane (Trypan Blue Assay), mitochondria (WST-1 assay), cellular proliferation (BrdU assay), and cell cycle (Propidium iodide and flow-cytometry). AgNPs showed surface plasmon resonance (SPR) of approximately 408 nm and average size of 3 nm. The starch-capped AgNPs successfully induced damage in cytoplasmic membrane and mitochondria, at concentrations equal and above 20 ppm. These damages lead to cell cycle arrest in G0/G1 and G2/M, blockage of proliferation and death in LNCaP and PC-3 cells, respectively. This data shows these AgNPs' potential as anticancer agents for the different stages of PC.

Keywords: anticancer agents; cytotoxicity; green synthesis; prostate cancer; silver nanoparticles; starch.

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Conflict of interest statement

All authors certify that they have no affiliations with or involvement in any organization or entity with any financial interest or non-financial interest in the subject matter or materials discussed in this manuscript.

Figures

Figure 1
Figure 1
(A) Photograph of starch capped AgNPs’ suspension; (B) UV-Vis spectrum of starch capped AgNPs’ suspension; (C) TEM image of AgNPs (20 nm resolution); (D) TEM image of AgNPs (5 nm resolution); (E) HRTEM image of AgNPs; (F) FFT from the blue box in Figure 1E showing spacings of ~2.3 and ~2.0 A.
Figure 2
Figure 2
Bright field microscopy images of LNCaP cell morphology after exposure to different concentrations of AgNPs ranging from 5 to 100 ppm, for 24 h and 48 h (20× OLYMPUS IX51 microscope).
Figure 3
Figure 3
Bright field microscopy images of PC-3 cell morphology after exposure to different concentrations of AgNPs ranging from 5 to 100 ppm, for 24 h and 48 h (20× OLYMPUS IX51 microscope).
Figure 4
Figure 4
LNCaP cells viability assessed by trypan blue exclusion method upon treatment with AgNPs for 24 h (A) and 48 h (B) and the comparison between their effect at the two time points (C). Results are expressed as percentage of control (untreated cells), as mean ± SEM.
Figure 5
Figure 5
PC-3 cells viability assessed by trypan blue exclusion method upon treatment with AgNPs for 24 h (A) and 48 h (B) and the comparison between their effect at the two time points (C). Results are expressed as percentage of control (untreated cells), as mean ± SEM.
Figure 6
Figure 6
Evaluation of cell viability, by WST-1 assay, upon LNCaP cells treatment with AgNPs at concentrations of 10–210 ppm for 24 h (A) and 48 h (B) and the comparison between their effect at the two time points (C). Results are expressed as percentage of control (untreated cells), as mean ± SEM.
Figure 7
Figure 7
Evaluation of cell viability, by WST-1 assay, upon PC-3 cells treatment with AgNPs at concentrations of 10–210 ppm for 24 h (A) and 48 h (B) and the comparison between their effect at the two time points (C). Results are expressed as percentage of control (untreated cells), as mean ± SEM.
Figure 8
Figure 8
Effect of AgNPs at concentrations of 10–210 ppm, on LNCaP cell proliferation assessed by BrdU incorporation assay after 24 h. Results are expressed as percentage of control (untreated cells) considered as 100%, as mean ± SEM.
Figure 9
Figure 9
Effect of AgNPs at concentrations of 10–210 ppm, on PC-3 cell proliferation assessed by BrdU incorporation assay after 24 h (A) and 48 of treatment (B). and the comparison between their effect at the two time points (C). Results are expressed as percentage of control (untreated cells) considered as 100%, as mean ± SEM.
Figure 10
Figure 10
Cell cycle analysis, of LNCaP cells treated with different concentrations of AgNPs for 24 h, assessed using a PI stain and flow cytometry. (A)—Representative DNA histograms. (B)—Quantitative analysis of PC-3 cells AgNPs treated. Data is expressed as mean ± SEM.
Figure 11
Figure 11
Cell cycle analysis, of LNCaP cells treated with different concentrations of AgNPs for 24 h, assessed using a PI stain and flow cytometry. (A)—Representative DNA histograms. (B)—Quantitative analysis of PC-3 cells AgNPs treated. Data is expressed as mean ± SEM.
Figure 12
Figure 12
Cell cycle analysis, of PC-3 cells treated with different concentrations of AgNPs for 24 h, assessed using a PI stain and flow cytometry. (A)—Representative DNA histograms. (B)—Quantitative analysis of PC-3 cells AgNPs treated. Data is expressed as mean ± SEM.
Figure 13
Figure 13
Cell cycle analysis, of PC-3 cells treated with different concentrations of AgNPs for 48 h, assessed using a PI stain and flow cytometry. (A)—Representative DNA histograms images. (B)—Quantitative analysis of PC-3 cells AgNPs treated. Data is expressed as mean ± SEM.
Figure 14
Figure 14
AgNPs’ cellular uptake quantification through Ag+ quantification by atomic absorption spectrometry (AAS) in LNCaP cells (A,B) and in PC-3 cells (C,D).
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