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An In Vitro Protocol for Evaluating MicroRNA Levels, Functions, and Associated Target Genes in Tumor Cells
Published on: May 21, 2019
Delta Np63 alpha – Responsive microRNA Modulate the Expression of Metabolic Enzymes
1Head and Neck Cancer Research Division, Cancer Research Building II, 1550 Orleans Street, Baltimore, MD 21231, USA. eratovi1@jhmi.edu.
Abstract:
MicroRNAs, whose transcription is regulated by members of the tumor protein p53 family, modulate the expression of numerous metabolic enzymes, significantly altering tumor cell response to chemotherapeutic treatments. The role for ΔNp63α-regulated microRNAs in regulation of cell cycle arrest, apoptosis and autophagy in squamous cell carcinoma (SCC) cells upon cisplatin exposure has been reported. The current study indicated that the selected microRNA targets differentially regulated by ΔNp63α in cisplatin-sensitive and cisplatin-resistant SCC cells could alter the expression of a few metabolic enzymes, thereby potentially contributing to the metabolic changes in SCC cells upon cisplatin exposure. Finally, the modulation of specific targets (e.g., SREBF2, AKT2, G6PD, CPS1, FADS1, and ETNK1) using a combination of microRNA mimics and siRNA silencing has shown that a suppression of these metabolic factors/ enzymes could confer a sensitivity of SCC cells to cisplatin. Thus, the Δ Np63α-regulated microRNAs were found to regulate the levels of several metabolic factors and enzymes, thereby potentially contributing to the response of larynx and tongue-derived SCC cells to platinum chemotherapy.
Insights
Delta Np63α-regulated microRNAs influence metabolic enzyme expression in squamous cell carcinoma (SCC). Suppressing these targets enhances SCC cell sensitivity to cisplatin chemotherapy, offering new therapeutic strategies.
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- MicroRNAs (miRNAs) regulated by the tumor protein p53 family impact cancer cell metabolism and drug response.
- Delta Np63α (ΔNp63α) plays a role in regulating cell cycle arrest, apoptosis, and autophagy in squamous cell carcinoma (SCC) upon cisplatin treatment.
- Differential regulation of miRNAs by ΔNp63α in cisplatin-sensitive versus resistant SCC cells suggests a role in metabolic reprogramming.
Purpose of the Study:
- To investigate how ΔNp63α-regulated microRNAs and their metabolic targets influence cisplatin response in SCC.
- To identify specific metabolic enzymes and factors modulated by ΔNp63α-regulated miRNAs in SCC cells.
- To determine if targeting these metabolic factors can restore cisplatin sensitivity in resistant SCC cells.
Main Methods:
- Differential expression analysis of microRNA targets in cisplatin-sensitive and resistant SCC cells.
- Utilizing microRNA mimics and siRNA silencing to modulate specific target genes (SREBF2, AKT2, G6PD, CPS1, FADS1, ETNK1).
- Assessing the impact of target modulation on SCC cell sensitivity to cisplatin chemotherapy.
Main Results:
- ΔNp63α-regulated microRNAs target specific metabolic enzymes, with differential regulation observed between cisplatin-sensitive and resistant SCC cells.
- Modulation of identified targets (SREBF2, AKT2, G6PD, CPS1, FADS1, ETNK1) significantly altered metabolic enzyme expression.
- Suppression of these specific metabolic targets using miRNA mimics and siRNA conferred cisplatin sensitivity to SCC cells.
Conclusions:
- ΔNp63α-regulated microRNAs are key regulators of metabolic reprogramming in SCC cells exposed to cisplatin.
- Targeting specific metabolic enzymes and factors identified in this study can overcome cisplatin resistance in SCC.
- These findings highlight a potential therapeutic strategy for enhancing platinum chemotherapy efficacy in larynx and tongue SCC.
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