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Optimized Automated Analysis of Live Neuronal Mitochondria Homeostasis Modulation by Isoform-Specific Retinoic Acid Receptors
Published on: July 28, 2023
[Proteomic study of retinoid acid resistant NB4R1 cells apoptosis induced by realgar]
Jun Qi1, Mei Zhang, Peng-Cheng He
1Department of Hematology, the First Clinical Hospital of Xi'an Jiaotong University, Xi'an 710061, China.
Objective:
To obtain and identify the differential proteome of apoptosis induced by realgar (tetra-arsenic tetra-sulfide, As(4)S(4)) in retinoid acid (RA) resistant human acute promyelocytic leukemia (APL) cell line NB4-R1 cells.
Methods:
The comparative proteomic expressive profiles of NB4-R1 cells treated with and without As(4)S(4) were obtained with the high-resolution two-dimensional electrophoresis system. After the analysis of ImageMaster(TM) 2D Platinum software combining artificial comparison. The differential expression proteins were screened and performed in-gel digestion and extraction of peptides, applied mass spectrometry MALDI-TOF-MS, MALDI-TOF/TOF, UPLC-MS/MS and bioinformatics to identify differential expressive proteins.
Results:
Twenty-two spots with more than 2-fold (≥ 2 or ≤ 0.5) expression changes were identified and 21 known proteins obtained, including 5 down-regulated (SET/I2PP2A, RPP2, PCBP1, EIF4H-1 and ANP32A/I1PP2A) after exposed to As(4)S(4), 2 up-regulated (HSP27 and HMGB1) after exposed for 24 h, and 14 up-regulated (PHB, ERP29, DNAJC8, PSMB4, ACTB, RPP0, RhoGDI2, alpha-tubulin, transcription factor, RBM15/OTT1, eIF5A1 and H2B1M et al.) after exposed for 48 h.
Conclusion:
It is the first time to successfully obtain and identify the global proteome of apoptosis induced by As(4)S(4) in RA-resistant cells. Among them, expressional and functional regulation of target proteins SET, RPP2 and PHB might be the potential novel therapeutic target for RA-resistant APL.
Insights
Realgar (As(4)S(4)) induces apoptosis in drug-resistant leukemia cells by altering protein expression. Key proteins like SET, RPP2, and PHB show altered expression, offering potential therapeutic targets for acute promyelocytic leukemia (APL).
Area of Science:
- Proteomics
- Cancer Biology
- Pharmacology
Background:
- Acute promyelocytic leukemia (APL) can develop resistance to retinoid acid (RA) treatment.
- Realgar (tetra-arsenic tetra-sulfide, As(4)S(4)) is investigated for its potential to induce apoptosis in resistant cancer cells.
Purpose of the Study:
- To identify the differential proteome associated with As(4)S(4)-induced apoptosis in RA-resistant APL cells (NB4-R1).
- To uncover potential therapeutic targets for overcoming RA resistance in APL.
Main Methods:
- Comparative proteomic analysis using high-resolution two-dimensional electrophoresis.
- Mass spectrometry (MALDI-TOF-MS, MALDI-TOF/TOF, UPLC-MS/MS) for protein identification.
- Bioinformatics analysis to interpret differential protein expression profiles.
Main Results:
- Twenty-two protein spots exhibited significant expression changes (≥ 2-fold) upon As(4)S(4) treatment.
- Identified 21 known proteins, including down-regulated proteins (e.g., SET/I2PP2A, RPP2) and up-regulated proteins (e.g., HSP27, HMGB1, PHB).
- Differential expression patterns were observed at 24 and 48 hours post-treatment.
Conclusions:
- This study provides the first global proteome analysis of As(4)S(4)-induced apoptosis in RA-resistant APL cells.
- The proteins SET, RPP2, and PHB, with their altered expression and function, represent potential novel therapeutic targets for RA-resistant APL.
- Findings suggest As(4)S(4) as a promising agent for targeting drug-resistant APL.
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