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Gold nanoparticles induce nuclear damage in breast cancer cells, which is further amplified by hyperthermia

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Abstract

Gold nanoparticles have emerged as promising tools for cancer research and therapy, where they can promote thermal killing. The molecular mechanisms underlying these events are not fully understood. The geometry and size of gold nanoparticles can determine the severity of cellular damage. Therefore, small and big gold nanospheres as well as gold nanoflowers were evaluated side-by-side. To obtain quantitative data at the subcellular and molecular level, we assessed how gold nanoparticles, either alone or in combination with mild hyperthermia, altered the physiology of cultured human breast cancer cells. Our analyses focused on the nucleus, because this organelle is essential for cell survival. We showed that all the examined gold nanoparticles associated with nuclei. However, their biological effects were quantitatively different. Thus, depending on the shape and size, gold nanoparticles changed multiple nuclear parameters. They redistributed stress-sensitive regulators of nuclear biology, altered the nuclear morphology, reorganized nuclear laminae and envelopes, and inhibited nucleolar functions. In particular, gold nanoparticles reduced the de novo biosynthesis of RNA in nucleoli, the subnuclear compartments that produce ribosomes. While small gold nanospheres and nanoflowers, but not big gold nanospheres, damaged the nucleus at normal growth temperature, several of these defects were further exacerbated by mild hyperthermia. Taken together, the toxicity of gold nanoparticles correlated with changes in nuclear organization and function. These results emphasize that the cell nucleus is a prominent target for gold nanoparticles of different morphologies. Moreover, we demonstrated that RNA synthesis in nucleoli provides quantitative information on nuclear damage and cancer cell survival.

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Abbreviations

BSA:

Bovine serum albumin

CAS:

Cellular apoptosis susceptibility protein

DAPI:

4′,6-Diamidino-2-phenylindole

GNP:

Gold nanoparticle

H3:

Histone H3

ICP-MS:

Inductively coupled plasma mass spectroscopy

LSP:

Localized surface plasmon resonance

MTT:

3-(4,5-dimethythiazol-2-yl)-2,5-Diphenyl tetrazolium bromide

NIR:

Near-infrared

NPC:

Nuclear pore complex

STDEV:

Standard deviation

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Acknowledgments

The authors are supported by grants from CIHR (Canadian Institutes of Health Research), NSERC (Natural Sciences and Engineering Council of Canada) and FQRNT (Fonds de Recherche du Québec–Nature et Technologies).

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Authors and Affiliations

  1. Department of Physiology, McGill University, Montreal, H3G 1Y6, Canada

    Mohamed Kodiha & Ursula Stochaj

  2. Department of Pharmacology and Therapeutics, McGill University, Montreal, H3G 1Y6, Canada

    Eliza Hutter, Sebastien Boridy, Michal Juhas & Dusica Maysinger

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  1. Mohamed Kodiha
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  2. Eliza Hutter
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  3. Sebastien Boridy
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  4. Michal Juhas
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Correspondence to Ursula Stochaj.

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18_2014_1622_MOESM1_ESM.tif

Suppl. Figure 1 Effect of gold nanospheres and nanoflowers on mitochondrial metabolic activity and proliferation. Dose–response curves are shown for MCF7 cells incubated with small (•) or big (●) gold nanospheres or gold nanoflowers (♦). MTT assays were carried out after 24-h incubation with GNPs; all results were normalized to untreated control cells. Each data point represents averages of at least three independent experiments performed in triplicates. Error bars indicate SEM. (B) Phosphorylation of histone H3. After overnight incubation with GNPs, MCF7 cells were further incubated for 3 h at 37 °C (No stress) or heat-shocked for 1 h at 43 °C and allowed to recover for 2 h or 24 h at 37 °C. Attached and floating cells were harvested and analyzed by quantitative Western blotting with antibodies against phospho(Ser10)-H3 and actin. For each data set, controls were incubated with vehicle. Results were normalized to controls, and changes in the ratio phospho-H3/actin are shown for two independent experiments. DNA synthesis was measured in controls or GNP-treated cells. EdU-positive cells were identified by quantitative image analysis. A minimum of 156 cells per data point was scored in each of two independent experiments. Results were normalized to vehicle-treated cells for each time point. V vehicle; S small gold nanospheres; F gold nanoflowers; B big gold nanospheres (TIFF 895 kb)

18_2014_1622_MOESM2_ESM.tif

Suppl. Figure 2 Detection of gold nanospheres in cytoplasmic and nuclear fractions. MCF7 cells were incubated overnight in the absence (V vehicle) or presence of small gold nanospheres (S). Cytoplasmic and nuclear fractions were prepared and GNPs detected by transmission electron microscopy (EM). The corresponding inductively coupled plasma mass spectroscopy (ICP-MS) values for cytoplasmic and nuclear fractions are also shown. Note that small gold nanospheres were present in both fractions (TIFF 520 kb)

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Kodiha, M., Hutter, E., Boridy, S. et al. Gold nanoparticles induce nuclear damage in breast cancer cells, which is further amplified by hyperthermia. Cell. Mol. Life Sci. 71, 4259–4273 (2014). https://doi.org/10.1007/s00018-014-1622-3

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