Apoptosis in human liver carcinoma caused by gold nanoparticles in combination with carvedilol is mediated via modulation of MAPK/Akt/mTOR pathway and EGFR/FAAD proteins

De Araújo Jr,Raimundo F.; Pessoa,Jonas B.; Cruz,Luis J.; Chan,Alan B.; De Castro Miguel,Emílio; Cavalcante,Rômulo S.; Brito,Gerly Anne C.; Silva,Heloiza Fernada O.; Gasparotto,Luiz H.S.; Guedes,Paulo M.M.; Araújo,Aurigena A.

doi:10.3892/ijo.2017.4179

Apoptosis in human liver carcinoma caused by gold nanoparticles in combination with carvedilol is mediated via modulation of MAPK/Akt/mTOR pathway and EGFR/FAAD proteins

Authors:
- Raimundo F. De Araújo Jr
- Jonas B. Pessoa
- Luis J. Cruz
- Alan B. Chan
- Emílio De Castro Miguel
- Rômulo S. Cavalcante
- Gerly Anne C. Brito
- Heloiza Fernada O. Silva
- Luiz H.S. Gasparotto
- Paulo M.M. Guedes
- Aurigena A. Araújo
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Affiliations: Department of Morphology, Federal University of Rio Grande do Norte, Natal 59072-970, RN, Brazil, Post Graduation Programme in Structural and Functional Biology, Federal University of Rio Grande do Norte, Natal 59072-970, RN, Brazil, Translational Nanobiomaterials and Imaging, Department of Radiology, Leiden University Medical Center, 2333 CL Leiden, The Netherlands, Percuros B.V., 2333 CL Leiden, The Netherlands, Department of Physical/Analytical Center/UFC, Fortaleza, CE, Brazil, Post Graduation Programme in Health Science, Federal University of Rio Grande do Norte, Natal 59072-970, RN, Brazil, Department of Morphology/Postgraduate Program in Morphology/UFC, Fortaleza, CE, Brazil, Group of Biological Chemistry and Chemometrics, Institute of Chemistry, Federal University of Rio Grande do Norte, Natal 59072-970, RN, Brazil, Department of Parasitology and Microbiology and Post Graduation Program in Parasitary Biology, Federal University of Rio Grande do Norte, Natal 59072-970, RN, Brazil, Department of Biophysics and Pharmacology, Post Graduation Programme in Public Health, Post Graduation Programme in Pharmaceutical Science, Federal University of Rio Grande do Norte, Natal 59072-970, RN, Brazil
Published online on: October 27, 2017 https://doi.org/10.3892/ijo.2017.4179
Pages: 189-200

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Abstract

In cancers, apoptosis signaling pathways and cell survival and growth pathways responsible for resistance to conventional treatments, such as Pi3K/Akt/mTOR and mitogen-activated protein kinase (MAPK) become dysregulated. Recently, alternative treatments to promote tumor cell death have become important. The present study reports on the antitumor and cytoprotective action of gold nanoparticles (GNPs) and carvedilol in combination and in isolated application. Apoptosis was analyzed by FITC/propidium iodide staining flow cytometry; caspase-3, caspase-8, Bcl-2 and MAPK/ERK activity by immunofluorescence microscopy; gene expression of proteins related to cell death as Akt, mTOR, EGFR, MDR1, survivin, FADD and Apaf, by the real-time PCR; and western blot analysis for MAPK/ERK, Akt and mTOR. Oxidative stress evaluation was performed by reduced glutathione (GSH) and malondialdehyde (MDA) levels. Intracellular GNPs targets were identified by transmission electron microscopy. After exposure to a combination of GNPs (6.25 µg/ml) and carvedilol (3 µM), death as promoted by apoptosis was detected using flow cytometry, for expression of pro-apoptotic proteins FADD, caspase-3, caspase-8 and sub-regulation of anti-apoptotic MAPK/ERK, Akt, mTOR, EGFR and MDR1 resistance. Non-tumor cell cytoprotection with GSH elevation and MDA reduction levels was detected. GNPs were identified within the cell near to the nucleus when combined with carvedilol. The combination of GNP and carvedilol promoted downregulation of anti-apoptotic and drug resistance genes, over-regulation of pro-apoptotic proteins in tumor cells, as well as cytoprotection of non-tumor cells with reduction of apoptosis and oxidative stress.

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