The Role of Insulin Receptor Substrate 4 (IRS4) Protein in the Radiotherapy Response of Glioblastoma Multiforme (GBM) Cells

Abstract

Objective: To explain the fundamental role of Insulin Receptor Substrate 4 (IRS4) protein in the response of Glioblastoma Multiforme (GBM) cells to radiotherapy. Methods: LN229 cells were transfected with IRS4 expression vector using lipofectamine, and the ectopic IRS4 expression was confirmed by western blot. After irradiating LN229 cells with 5, 8, and 10 Gy doses of radiotherapy, the functional effect of IRS4 on radiotherapy was determined using MTT and colony formation assays. Results: It was determined that increased IRS4 expression led to enhanced radiosensitivity in GBM cells. Increased IRS4 expression in the GBM cell line was found to cause a decrease in cell survival rates and colony formation rates. Conclusion: IRS4 has been identified to potentially play an active role in the radiotherapy response of GBM cells.

Keywords

• IRS4
• GBM
• Radiotherapy

References

1. 1. Witthayanuwat S, Pesee M, Supaadirek C, Supakalin N, Thamronganantasakul K, Krusun S. Survival analysis of glioblastoma multiforme. Asian Pac J Cancer Prev. 2018;19(9):2613–2617.
2. 2. Dağıstan Y, Dağıstan E, Güney MS. Sebepsiz kişilik değişiklikleri ile başlayan glioblastome multiforme: Bir olgu sunumu. Abant Tıp Derg. 2012;1(1):23-25.
3. 3. Qazi MA, Vora P, Venugopal C, et al. Intratumoral heterogeneity: pathways to treatment resistance and relapse in human glioblastoma. Ann Oncol. 2017;28(7):1448-1456.
4. 4. Burri SH, Gondi V, Brown PD, Mehta MP. The Evolving Role of Tumor Treating Fields in Managing Glioblastoma: Guide for Oncologists. Am J Clin Oncol. 2018;41(2):191-196.
5. 5. Delgado-Martin B, Medina MA. Advances in the Knowledge of the Molecular Biology of Glioblastoma and Its Impact in Patient Diagnosis, Stratification, and Treatment. Adv Sci (Weinh). 2020;7(9):1902971.
6. 6. Pastwa E, Neumann RD, Mezhevaya K, Winters TA. Repair of radiation-induced DNA double-strand breaks is dependent upon radiation quality and the structural complexity of double-strand breaks. Radiat Res. 2003;159(2):251-261.
7. 7. Qiu GH. Protection of the genome and central protein-coding sequences by non-coding DNA against DNA damage from radiation. Mutat Res Rev Mutat Res. 2015;764:108-117.
8. 8. Prasanna PG, Stone HB, Wong RS, et al. Normal tissue protection for improving radiotherapy: Where are the Gaps? Transl Cancer Res. 2012;1(1):35-48.

9. 9. Gong Y, Ma Y, Sinyuk M, Loganathan S, et al. Insulin-mediated signaling promotes proliferation and survival of glioblastoma through Akt activation. Neuro Oncol. 2016,18(1):48-57.
10. 10. Machado-Neto JA, Traina F. IRS1 (insulin receptor substrate 1). Atlas Genet Cytogenet Oncol Hematol. 2013;17(9):594-598.
11. 11. Sanmartín-Salinas P, Toledo-Lobo MV, Noguerales-Fraguas F, et al. Overexpression of insulin receptor substrate-4 is correlated with clinical staging in colorectal cancer patients. J Mol Histol. 2018;49(1):39-49.
12. 12. Hoxhaj G, Dissanayake K, MacKintosh C. Effect of IRS4 levels on PI3-kinase signalling. PLoS One. 2013;8(9):e73327.
13. 13. Malaguarnera R, Belfiore AThe insulin receptor: a new target for cancer therapy. Front Endocrinol (Lausanne). 2011;2:93.
14. 14. Mendoza MC, Er EE, Blenis J. The Ras-ERK and PI3K-mTOR pathways: cross-talk and compensation. Trends Biochem Sci. 2011;36(6):320-328.
15. 15. Sesti G, Federici M, Hribal ML, Lauro D, Sbraccia P, Lauro R. Defects of the insulin receptor substrate (IRS) system in human metabolic disorders. FASEB J. 2001;15(12):2099-111.
16. 16. Trojanek J, Ho T, Del Valle L, et al. Role of the insulin-like growth factor I/insulin receptor substrate 1 axis in Rad51 trafficking and DNA repair by homologous recombination. Mol Cell Biol. 2003,23(21):7510-7524.
17. 17. Trojanek J, Croul S, Ho T, et al. T-antigen of the human polyomavirus JC attenuates faithful DNA repair by forcing nuclear interaction between IRS-1 and Rad51. J Cell Physiol. 2006;06(1):35-46.
18. 18. Dörpholz G, Murgai A, Jatzlau J, et al. IRS4, a novel modulator of BMP/Smad and Akt signalling during early muscle differentiation. Sci Rep. 2017;7(1):8778.