Proteomic and metabolomic profiling combined with in vitro studies reveal the antiproliferative mechanism of silver nanoparticles in MDA-MB-231 breast carcinoma cells

• Published in: Journal of materials chemistry. B, Materials for biology and medicine Vol. 10; no. 13; pp. 2148 - 2159
• Main Authors: Nirmala, J Grace, Meher, Kimaya, Lopus, Manu
• Format: Journal Article
• Language: English
• Published: England Royal Society of Chemistry 30.03.2022
• Subjects: Antiproliferatives; Apoptosis; Binding; Breast cancer; Breast carcinoma; Breast Neoplasms - drug therapy; Cell cycle; Cell death; Cell Line, Tumor; Cell Survival; Cytotoxicity; Disruption; Female; Flow cytometry; Humans; Immunofluorescence; Lipid metabolism; Lipids; Metabolism; Metabolomics; Metal Nanoparticles - chemistry; Microtubules; Nanoparticles; Polymers; Proteomics; Silver; Silver - chemistry; Silver - pharmacology; Toxicity; Transmission electron microscopy; Tubulin
• ISSN: 2050-750X; 2050-7518
• DOI: 10.1039/d1tb02760c

Silver nanoparticles, shaped and stabilized by various means, are known to alter biological systems and promote cytotoxicity. However, the precise mechanism by which they induce toxic outcomes in cancer cells is poorly understood. Using a combination of cellular and biophysical assays and proteomic and metabolomic analyses, we report the cytotoxic mechanism of action of tryptone-stabilized silver nanoparticles (T-AgNPs). After their facile synthesis and characterization using an assortment of spectroscopic techniques and transmission electron microscopy, the mechanism of action of the particles was elucidated using MDA-MB-231 breast cancer cells as the cell model. The nanoparticles inhibited the proliferative (IC :100 ± 3 μg mL ) and clonogenic potential of the cells. Flow cytometry analyses revealed an absence of phase-specific cell cycle arrest but extensive cell death in the treated cells. The mechanism of action of the particles consisted of their direct binding to the microtubule-building protein tubulin and the disruption of its helical integrity, as confirmed fluorometric analysis and far-UV spectropolarimetry, respectively. The binding hampered the assembly of microtubules, as confirmed polymer mass analysis of assembled, purified tubulin and immunofluorescence imaging of cellular microtubules. Proteomic and metabolomic analyses revealed the downregulation of lipid metabolism to be a synergistic contributor to cell death. Taken together, we report a novel antiproliferative mechanism of action of T-AgNPs that involves tubulin disruption and the downregulation of lipid metabolism.