Methotrexate-associated Zinc Oxide Nanoparticles Reduce Cell Viability and Alters TP53 and COX-2 Gene Expressions in MCF7 Breast Cancer Cells
Abstract
BACKGROUND: Conventional breast cancer treatment faces a lot of challenges due to treatment resistance and undesirable side effects. Oxidative stress pathway is involved in the response of cancer cells to chemotherapeutic agents and nanomaterials, which is usually related to alteration of malondialdehyde (MDA), tumor protein p53 (TP53), and Cyclooxygenase-2 (COX-2). Methotrexate (MTX) has been proved to have anticancer effect, but its clinical use is limited. Associating it with zinc oxide (ZnO) could offer a means of enhancing the cellular effects of MTX. This study was conducted to synthesize MTX–ZnO nanoparticles (NPs) and evaluate their antiproliferative activity, oxidative stress, and TP53 and COX-2 gene expression in MCF7 breast cancer cells.
METHODS: MTX–ZnO NPs were synthesized and characterized using UV–Vis spectroscopy, dynamic light scattering, zeta potential analysis, scanning electron microscopy, and X-ray diffraction analysis. The MTT assay was used to evaluate the cytotoxic activity against MCF7 cells, enzyme-linked immunosorbent assay (ELISA) to evaluate MDA level, and quantitative real-time polymerase chain reaction (PCR) to evaluate TP53 and COX-2 expressions.
RESULTS: The synthesized MTX–ZnO NPs exhibited average hydrodynamic diameter of 56.8 nm, zeta potential of −21.9 mV, and predominantly spherical morphology. MTX–ZnO NPs showed a concentration dependent cytotoxicity, reducing MCF7 cell viability from 91% to 51%. Treatment with MTX–ZnO NPs resulted in increase of MDA levels (16.8 µM vs. 3.4 µM), suggesting the improvement of lipid peroxidation and oxidative stress after MTX–ZnO NPs treatment. In addition, TP53 expression increased from approximately 1.5-fold to 10.5-fold and COX-2 expression increased from approximately 3-fold to 57-fold.
CONCLUSION: MTX–ZnO NPs exhibited antiproliferative effects on MCF7 breast cancer cells and were associated with increased oxidative stress and upregulated TP53 and COX-2 expression. The results indicate the potential of MTX–ZnO NPs in breast cancer treatment researches.
KEYWORDS: methotrexate, zinc oxide nanoparticles, breast cancer, MCF7 cells, oxidative stress, TP53, COX-2
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DOI: https://doi.org/10.18585/inabj.v18i5.4330
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