Affiliation:
1. University of Tabriz
2. Tabriz University of Medical Sciences
Abstract
Abstract
Background
Gastric cancer (GC) is a highly chemoresistant malignancy with a poor prognosis. Paclitaxel's low response rate as second-line chemotherapy for advanced GC has prompted intensive research into its molecular basis and prospective targeted therapies to enhance its therapeutic efficacy. The objective of this study was to investigate the synergistic effects of NFE2L2 silencing in combination with paclitaxel treatment on GC cell viability, apoptosis, proliferation, autophagy, and migration.
Methods
After the siRNA-mediated silencing of NFE2L2 in AGS cells, the transfection efficacy was evaluated by qRT-PCR. The MTT assay was then applied to assess cell viability, followed by flow cytometry analysis for apoptosis, proliferation, and autophagy in AGS cells treated with NFE2L2 siRNA, paclitaxel, or their combination. Thereafter, the migration of cells was measured using a wound-healing assay. Ultimately, the relative gene expression levels of apoptotic (Bax, CASP3, and CASP9), anti-apoptotic (BCL2), metastatic (MMP2), and cell cycle (TP53) genes were measured by qRT-PCR in all experiment groups to further assess the molecular basis for the combination therapy.
Results
NFE2L2 siRNA transfection significantly enhanced paclitaxel-induced apoptosis and sensitized AGS cells to paclitaxel via modulating the expression of apoptosis-related genes including BCL2, Bax, CASP3, and CASP9. Besides, NFE2L2 siRNA and paclitaxel synergistically induced cell cycle arrest at the G2 phase, promoted autophagy activation, and inhibited AGS cell migration via MMP2 downregulation. Additionally, TP53, a key regulator of cell growth, was significantly upregulated in the treated groups compared to the control group.
Conclusions
Our findings suggest that paclitaxel combined with siRNA-mediated silencing of NFE2L2 might represent a promising therapeutic strategy for GC, however further translational and clinical research are warranted.
Publisher
Research Square Platform LLC
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