Anticancer activity of biogenerated silver nanoparticles: an integrated proteomic investigation
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Miriam Buttacavoli1,*, Nadia Ninfa Albanese1,*, Gianluca Di Cara2, Rosa Alduina1, Claudia Faleri3, Michele Gallo4, Giuseppe Pizzolanti5,6, Giuseppe Gallo1,6, Salvatore Feo1,6, Franco Baldi4 and Patrizia Cancemi1,2,6
1Department of Biological Chemical and Pharmaceutical Sciences and Technologies (STEBICEF), University of Palermo, Palermo, Italy
2Center of Experimental Oncobiology (C.OB.S.), La Maddalena Hospital III Level Oncological Dept., Palermo, Italy
3Department of Life Science, University of Siena, Siena, Italy
4Department of Molecular Science and Nanosystems, Cà Foscari University of Venice, Venice, Italy
5Biomedical Department of Internal and Specialist Medicine (DIBIMIS), Section of Endocrinology, University of Palermo, Palermo, Italy
6Advanced Technologies Network Center (ATeN), University of Palermo, Palermo, Italy
*These authors contributed equally to this work
Patrizia Cancemi, email: [email protected]
Keywords: silver nanoparticles (AgNPs); bacteria; breast cancer cells; anticancer activity; proteomics
Received: May 27, 2017 Accepted: October 30, 2017 Published: December 23, 2017
Silver nanoparticles (AgNPs), embedded into a specific polysaccharide (EPS), were biogenerated by Klebsiella oxytoca DSM 29614 under aerobic (AgNPs-EPSaer) and anaerobic conditions (AgNPs-EPSanaer). Both AgNPs-EPS matrices were tested by MTT assay for cytotoxic activity against human breast (SKBR3 and 8701-BC) and colon (HT-29, HCT 116 and Caco-2) cancer cell lines, revealing AgNPs-EPSaer as the most active, in terms of IC50, with a more pronounced efficacy against breast cancer cell lines. Therefore, colony forming capability, morphological changes, generation of reactive oxygen species (ROS), induction of apoptosis and autophagy, inhibition of migratory and invasive capabilities and proteomic changes were investigated using SKBR3 breast cancer cells with the aim to elucidate AgNPs-EPSaer mode of action. In particular, AgNPs-EPSaer induced a significant decrease of cell motility and MMP-2 and MMP-9 activity and a significant increase of ROS generation, which, in turn, supported cell death mainly through autophagy and in a minor extend through apoptosis. Consistently, TEM micrographs and the determination of total silver in subcellular fractions indicated that the Ag+ accumulated preferentially in mitochondria and in smaller concentrations in nucleus, where interact with DNA. Interestingly, these evidences were confirmed by a differential proteomic analysis that highlighted important pathways involved in AgNPs-EPSaer toxicity, including endoplasmic reticulum stress, oxidative stress and mitochondrial impairment triggering cell death trough apoptosis and/or autophagy activation.
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