Inhibition of NUPR1–Karyopherin β1 Binding Increases Anticancer Drug Sensitivity
Abstract
:1. Introduction
2. Results
2.1. Dose- and Time-Dependent Cytotoxic Effects of Doxorubicin on MDA-MB 231 Breast Cancer Cells
2.2. Increased NUPR1 Nuclear Transportation after Doxorubicin Treatment on MDA-MB-231 Breast Cancer Cells
2.3. ATZ-502 Inhibits NUPR1 Activity in MDA-MB-231 Breast Cancer Cells by Inhibiting Nuclear Transportation
2.4. Inhibition of NUPR1 Nucleus Transportation Induces G2/M Phase Arrest and Increases Synergistic Cytotoxicity Effects
2.5. RNA Quant-Seq Analysis Revealed That Single and Combination Treatments Yielded Different mRNA Expression Patterns
2.6. Gene Ontology Analysis for DEGs
2.7. NUPR1-Induced Gene Expression Is Differentially Regulated between Single-Treatment and Combination-Treatment Groups
2.8. Karyopherin β1 Binds to NUPR1 with High Affinity
2.9. ATZ-502 Inhibits the NUPR1–Karyopherin β1 Interaction
3. Discussion
4. Materials and Methods
4.1. Cell Culture
4.2. Cell Viability Assay
4.3. Cell Cycle and Apoptosis with Fluorescence-Activated Cell Sorting (FACS) Analysis
4.4. Single-Molecule Binding Assay
4.5. Drug Inhibition Assay at the Single-Molecule Level
4.6. Immunofluorescence Assay
4.7. Gene Expression Profiling Analysis Using QuantSeq 3′ mRNA-seq
4.8. Data Analysis and Gene Selection
4.9. Quantitative Real-Time RT-PCR
4.10. Protein Expression, Immunoprecipitaton, and Western Blotting
4.11. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Cell Lines | ATZ-50 (μM) | IC50 (μM) | Fold Difference | |
---|---|---|---|---|
Single | Combination | |||
MDA-MB-231 | 0.50 | 1.767 | 0.438 | 4.0 |
SiHa | 0.50 | 0.624 | 0.239 | 2.6 |
MCF-7/ADR | 0.05 | 39.62 | 2.00 | 19.8 |
SK-OV-3 | 0.01 | 0.250 | 0.175 | 1.4 |
PC-3 | 0.01 | 0.357 | 0.018 | 19.6 |
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Park, C.; Oh, J.; Lee, W.M.; Koh, H.R.; Sohn, U.D.; Ham, S.W.; Oh, K. Inhibition of NUPR1–Karyopherin β1 Binding Increases Anticancer Drug Sensitivity. Int. J. Mol. Sci. 2021, 22, 2794. https://doi.org/10.3390/ijms22062794
Park C, Oh J, Lee WM, Koh HR, Sohn UD, Ham SW, Oh K. Inhibition of NUPR1–Karyopherin β1 Binding Increases Anticancer Drug Sensitivity. International Journal of Molecular Sciences. 2021; 22(6):2794. https://doi.org/10.3390/ijms22062794
Chicago/Turabian StylePark, Chanhee, Jiwon Oh, Won Mo Lee, Hye Ran Koh, Uy Dong Sohn, Seung Wook Ham, and Kyungsoo Oh. 2021. "Inhibition of NUPR1–Karyopherin β1 Binding Increases Anticancer Drug Sensitivity" International Journal of Molecular Sciences 22, no. 6: 2794. https://doi.org/10.3390/ijms22062794