Overexpression of Insulin Receptor Substrate 1 (IRS1) Relates to Poor Prognosis and Promotes Proliferation, Stemness, Migration, and Oxidative Stress Resistance in Cholangiocarcinoma
Abstract
:1. Introduction
2. Results
2.1. IRS1 Expression Profiles in CCA Patients in Relation to Clinicopathological Features and 8-Oxodg Formation
2.2. Expression of IRS1 in Cells Exposed Long-Term to H2O2
2.3. Transient IRS1 Knockdown in CCA Cell Lines
2.4. Transient IRS1 Knockdown Suppressed Proliferation of CCA Cell Lines
2.5. Transient IRS1 Knockdown Induced G1 Cell Cycle Arrest of CCA Cell Lines
2.6. Transient IRS1 Knockdown Reduced Stem-like Properties of CCA Cell Lines
2.7. Transient IRS1 Knockdown Decreased Migration and Invasion Activities of CCA Cell Lines
2.8. Transient IRS1 Knockdown Reduced Oxidative Stress-Resistant Property of CCA Cell Lines
3. Discussion
4. Materials and Methods
4.1. Human Intrahepatic CCA Tissues
4.2. Immunohistochemistry
4.3. Cell Culture
4.4. Real-Time PCR
4.5. Immunocytochemistry
4.6. Western Blot Analysis
4.7. Small Interfering RNA (siRNA) Transfection
4.8. Hydrogen Peroxide Treatment
4.9. Carbonylated Protein Determination
4.10. Cell Proliferation Assay
4.11. Cell Migration Assay
4.12. Cell Invasion Assay
4.13. Cell Cycle Arrest Assay
4.14. RNA Sequencing
4.15. Spheroid Formation
4.16. Data analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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IRS1 Protein Expression | Statistical Analysis | ||
---|---|---|---|
Low (n = 31) | High (n = 53) | ||
Survival days (Mean ± SD) | 480 ± 342 | 301 ± 218 | p = 0.007 # |
Age (≤57/>57) | 15/16 | 26/27 | p = 0.953 * |
Gender (Male/Female) | 21/10 | 40/13 | p = 0.443 * |
Histology types | |||
Tubular/Papillary | 17/14 | 32/21 | p = 0.619 * |
Metastasis | |||
No/Yes | 17/14 | 18/35 | p = 0.061 * |
8-oxodG formation | |||
Low/High | 21/10 | 16/37 | p < 0.001 * |
IRS1 and 8-oxodG Protein Expression | Statistical Analysis * | |||
---|---|---|---|---|
Low IRS1 and 8-oxodG (n = 21) | High IRS1 or 8-oxodG (n = 26) | High IRS1 and 8-oxodG (n = 37) | ||
Age (≤57/>57) | 10/11 | 12/14 | 19/18 | p = 0.913 |
Gender (male/female) | 16/5 | 18/8 | 27/10 | p = 0.866 |
Histology types | ||||
Tubular/Papillary | 8/13 | 17/9 | 24/13 | p = 0.094 |
Metastasis | ||||
No/Yes | 14/7 | 8/8 | 13/24 | p = 0.026 |
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Kaewlert, W.; Sakonsinsiri, C.; Lert-itthiporn, W.; Ungarreevittaya, P.; Pairojkul, C.; Pinlaor, S.; Murata, M.; Thanan, R. Overexpression of Insulin Receptor Substrate 1 (IRS1) Relates to Poor Prognosis and Promotes Proliferation, Stemness, Migration, and Oxidative Stress Resistance in Cholangiocarcinoma. Int. J. Mol. Sci. 2023, 24, 2428. https://doi.org/10.3390/ijms24032428
Kaewlert W, Sakonsinsiri C, Lert-itthiporn W, Ungarreevittaya P, Pairojkul C, Pinlaor S, Murata M, Thanan R. Overexpression of Insulin Receptor Substrate 1 (IRS1) Relates to Poor Prognosis and Promotes Proliferation, Stemness, Migration, and Oxidative Stress Resistance in Cholangiocarcinoma. International Journal of Molecular Sciences. 2023; 24(3):2428. https://doi.org/10.3390/ijms24032428
Chicago/Turabian StyleKaewlert, Waleeporn, Chadamas Sakonsinsiri, Worachart Lert-itthiporn, Piti Ungarreevittaya, Chawalit Pairojkul, Somchai Pinlaor, Mariko Murata, and Raynoo Thanan. 2023. "Overexpression of Insulin Receptor Substrate 1 (IRS1) Relates to Poor Prognosis and Promotes Proliferation, Stemness, Migration, and Oxidative Stress Resistance in Cholangiocarcinoma" International Journal of Molecular Sciences 24, no. 3: 2428. https://doi.org/10.3390/ijms24032428