Nuclear IGF1R interacts with NuMA and regulates 53BP1‑dependent DNA double‑strand break repair in colorectal cancer

Yang,Chen; Zhang,Yifan; Segar,Nelly; Huang,Changhao; Zeng,Pengwei; Tan,Xiangzhou; Mao,Linfeng; Chen,Zhikang; Haglund,Felix; Larsson,Olle; Chen,Zihua; Lin,Yingbo

doi:10.3892/or.2021.8119

Nuclear IGF1R interacts with NuMA and regulates 53BP1‑dependent DNA double‑strand break repair in colorectal cancer

Authors:
- Chen Yang
- Yifan Zhang
- Nelly Segar
- Changhao Huang
- Pengwei Zeng
- Xiangzhou Tan
- Linfeng Mao
- Zhikang Chen
- Felix Haglund
- Olle Larsson
- Zihua Chen
- Yingbo Lin
View Affiliations

Affiliations: Department of General Surgery, Xiangya Hospital of Central South University, Changsha, Hunan 410000, P.R. China, Department of Clinical Pathology and Cytology, Karolinska University Hospital Solna, 171 64 Solna, Stockholm, Sweden, Department of Oncology and Pathology, Karolinska Institute, 171 77 Stockholm, Sweden
Published online on: June 22, 2021 https://doi.org/10.3892/or.2021.8119
Article Number: 168
Copyright: © Yang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Nuclear insulin‑like growth factor 1 receptor (nIGF1R) has been associated with poor overall survival and chemotherapy resistance in various types of cancer; however, the underlying mechanism remains unclear. In the present study, immunoprecipitation‑coupled mass spectrometry was performed in an IGF1R‑overexpressing SW480‑OE colorectal cancer cell line to identify the nIGF1R interactome. Network analysis revealed 197 proteins of interest which were involved in several biological pathways, including RNA processing, DNA double‑strand break (DSB) repair and SUMOylation pathways. Nuclear mitotic apparatus protein (NuMA) was identified as one of nIGF1R's colocalizing partners. Proximity ligation assay (PLA) revealed different levels of p53‑binding protein 1 (53BP1)‑NuMA colocalization between IGF1R‑positive (R⁺) and IGF1R‑negative (R^‑) mouse embryonic fibroblasts following exposure to ionizing radiation (IR). 53BP1 was retained by NuMA in the R^‑ cells during IR‑induced DNA damage. By contrast, the level of NuMA‑53BP1 was markedly lower in R⁺ cells compared with R^‑ cells. The present data suggested a regulatory role of nIGF1R in 53BP1‑dependent DSB repair through its interaction with NuMA. Bright‑field PLA analysis on a paraffin‑embedded tissue microarray from patients with colorectal cancer revealed a significant association between increased nuclear colocalizing signals of NuMA‑53BP1 and a shorter overall survival. These results indicate that nIGF1R plays a role in facilitating 53BP1‑dependent DDR by regulating the NuMA‑53BP1 interaction, which in turn might affect the clinical outcome of patients with colorectal cancer.

View Figures

View References

1	Dekker E, Tanis PJ, Vleugels JLA, Kasi PM and Wallace MB: Colorectal cancer. Lancet. 394:1467–1480. 2019. View Article : Google Scholar : PubMed/NCBI
2	Mahar AL, Compton C, Halabi S, Hess KR, Weiser MR and Groome PA: Personalizing prognosis in colorectal cancer: A systematic review of the quality and nature of clinical prognostic tools for survival outcomes. J Surg Oncol. 116:969–982. 2017. View Article : Google Scholar : PubMed/NCBI
3	Fakih MG: Metastatic colorectal cancer: Current state and future directions. J Clin Oncol. 33:1809–1824. 2015. View Article : Google Scholar : PubMed/NCBI
4	Martini G, Troiani T, Cardone C, Vitiello P, Sforza V, Ciardiello D, Napolitano S, Della Corte CM, Morgillo F, Raucci A, et al: Present and future of metastatic colorectal cancer treatment: A review of new candidate targets. World J Gastroenterol. 23:4675–4688. 2017. View Article : Google Scholar : PubMed/NCBI
5	Dyer AH, Vahdatpour C, Sanfeliu A and Tropea D: The role of insulin-like growth factor 1 (IGF-1) in brain development, maturation and neuroplasticity. Neuroscience. 325:89–99. 2016. View Article : Google Scholar : PubMed/NCBI
6	Lin Y, Liu H, Waraky A, Haglund F, Agarwal P, Jernberg-Wiklund H, Warsito D and Larsson O: SUMO-modified insulin-like growth factor 1 receptor (IGF-1R) increases cell cycle progression and cell proliferation. J Cell Physiol. 232:2722–2730. 2017. View Article : Google Scholar : PubMed/NCBI
7	Heidegger I, Kern J, Ofer P, Klocker H and Massoner P: Oncogenic functions of IGF1R and INSR in prostate cancer include enhanced tumor growth, cell migration and angiogenesis. Oncotarget. 5:2723–2735. 2014. View Article : Google Scholar : PubMed/NCBI
8	Riedemann J and Macaulay VM: IGF1R signalling and its inhibition. Endocr Relat Cancer. 13 (Suppl 1):S33–S43. 2006. View Article : Google Scholar : PubMed/NCBI
9	Rodrigues Alves APN, Fernandes JC, Fenerich BA, Coelho-Silva JL, Scheucher PS, Simões BP, Rego EM, Ridley AJ, Machado-Neto JA and Traina F: IGF1R/IRS1 targeting has cytotoxic activity and inhibits PI3K/AKT/mTOR and MAPK signaling in acute lymphoblastic leukemia cells. Cancer Lett. 456:59–68. 2019. View Article : Google Scholar : PubMed/NCBI
10	Zorea J, Prasad M, Cohen L, Li N, Schefzik R, Ghosh S, Rotblat B, Brors B and Elkabets M: IGF1R upregulation confers resistance to isoform-specific inhibitors of PI3K in PIK3CA-driven ovarian cancer. Cell Death Dis. 9:9442018. View Article : Google Scholar : PubMed/NCBI
11	Warsito D, Lin Y, Gnirck AC, Sehat B and Larsson O: Nuclearly translocated insulin-like growth factor 1 receptor phosphorylates histone H3 at tyrosine 41 and induces SNAI2 expression via Brg1 chromatin remodeling protein. Oncotarget. 7:42288–42302. 2016. View Article : Google Scholar : PubMed/NCBI
12	Packham S, Warsito D, Lin Y, Sadi S, Karlsson R, Sehat B and Larsson O: Nuclear translocation of IGF-1R via p150(Glued) and an importin-β/RanBP2-dependent pathway in cancer cells. Oncogene. 34:2227–2238. 2015. View Article : Google Scholar : PubMed/NCBI
13	Warsito D, Sjostrom S, Andersson S, Larsson O and Sehat B: Nuclear IGF1R is a transcriptional co-activator of LEF1/TCF. EMBO Rep. 13:244–250. 2012. View Article : Google Scholar : PubMed/NCBI
14	Sehat B, Tofigh A, Lin Y, Trocmé E, Liljedahl U, Lagergren J and Larsson O: SUMOylation mediates the nuclear translocation and signaling of the IGF-1 receptor. Sci Signal. 3:ra102010. View Article : Google Scholar : PubMed/NCBI
15	Aleksic T, Verrill C, Bryant RJ, Han C, Worrall AR, Brureau L, Larré S, Higgins GS, Fazal F, Sabbagh A, et al: IGF-1R associates with adverse outcomes after radical radiotherapy for prostate cancer. Br J Cancer. 117:1600–1606. 2017. View Article : Google Scholar : PubMed/NCBI
16	Wang Y, Yuan JL, Zhang YT, Ma JJ, Xu P, Shi CH, Zhang W, Li YM, Fu Q, Zhu GF, et al: Inhibition of both EGFR and IGF1R sensitized prostate cancer cells to radiation by synergistic suppression of DNA homologous recombination repair. PLoS One. 8:e687842013. View Article : Google Scholar : PubMed/NCBI
17	Guerard M, Robin T, Perron P, Hatat AS, David-Boudet L, Vanwonterghem L, Busser B, Coll JL, Lantuejoul S, Eymin B, et al: Nuclear translocation of IGF1R by intracellular amphiregulin contributes to the resistance of lung tumour cells to EGFR-TKI. Cancer Lett. 420:146–155. 2018. View Article : Google Scholar : PubMed/NCBI
18	Her J and Bunting SF: How cells ensure correct repair of DNA double-strand breaks. J Biol Chem. 293:10502–10511. 2018. View Article : Google Scholar : PubMed/NCBI
19	Wright WD, Shah SS and Heyer WD: Homologous recombination and the repair of DNA double-strand breaks. J Biol Chem. 293:10524–10535. 2018. View Article : Google Scholar : PubMed/NCBI
20	Chang HHY, Pannunzio NR, Adachi N and Lieber MR: Non-homologous DNA end joining and alternative pathways to double-strand break repair. Nat Rev Mol Cell Biol. 18:495–506. 2017. View Article : Google Scholar : PubMed/NCBI
21	Escribano-Diaz C, Orthwein A, Fradet-Turcotte A, Xing M, Young JT, Tkáč J, Cook MA, Rosebrock AP, Munro M, Canny MD, et al: A cell cycle-dependent regulatory circuit composed of 53BP1-RIF1 and BRCA1-CtIP controls DNA repair pathway choice. Mol Cell. 49:872–883. 2013. View Article : Google Scholar : PubMed/NCBI
22	Chapman JR, Barral P, Vannier JB, Borel V, Steger M, Tomas-Loba A, Sartori AA, Adams IR, Batista FD and Boulton SJ: RIF1 is essential for 53BP1-dependent nonhomologous end joining and suppression of DNA double-strand break resection. Mol Cell. 49:858–871. 2013. View Article : Google Scholar : PubMed/NCBI
23	Salvador Moreno N, Liu J, Haas KM, Parker LL, Chakraborty C, Kron SJ, Hodges K, Miller LD, Langefeld C, Robinson PJ, et al: The nuclear structural protein NuMA is a negative regulator of 53BP1 in DNA double-strand break repair. Nucleic Acids Res. 47:2703–2715. 2019. View Article : Google Scholar : PubMed/NCBI
24	Chitnis MM, Lodhia KA, Aleksic T, Gao S, Protheroe AS and Macaulay VM: IGF-1R inhibition enhances radiosensitivity and delays double-strand break repair by both non-homologous end-joining and homologous recombination. Oncogene. 33:5262–5273. 2014. View Article : Google Scholar : PubMed/NCBI
25	Waraky A, Lin Y, Warsito D, Haglund F, Aleem E and Larsson O: Nuclear insulin-like growth factor 1 receptor phosphorylates proliferating cell nuclear antigen and rescues stalled replication forks after DNA damage. J Biol Chem. 292:18227–18239. 2017. View Article : Google Scholar : PubMed/NCBI
26	Doncheva NT, Morris JH, Gorodkin J and Jensen LJ: Cytoscape StringApp: Network analysis and visualization of proteomics data. J Proteome Res. 18:623–632. 2019. View Article : Google Scholar : PubMed/NCBI
27	Bindea G, Mlecnik B, Hackl H, Charoentong P, Tosolini M, Kirilovsky A, Fridman WH, Pagès F, Trajanoski Z and Galon J: ClueGO: A Cytoscape plug-in to decipher functionally grouped gene ontology and pathway annotation networks. Bioinformatics. 25:1091–1093. 2009. View Article : Google Scholar : PubMed/NCBI
28	Huang C, Yuan W, Lai C, Zhong S, Yang C, Wang R, Mao L and Chen Z and Chen Z: EphA2-to-YAP pathway drives gastric cancer growth and therapy resistance. Int J Cancer. 146:1937–1949. 2020. View Article : Google Scholar : PubMed/NCBI
29	ten Have S, Boulon S, Ahmad Y and Lamond AI: Mass spectrometry-based immuno-precipitation proteomics-the user's guide. Proteomics. 11:1153–1159. 2011. View Article : Google Scholar : PubMed/NCBI
30	Kakarougkas A and Jeggo PA: DNA DSB repair pathway choice: An orchestrated handover mechanism. Br J Radiol. 87:201306852014. View Article : Google Scholar : PubMed/NCBI
31	Xu Y and Her C: Inhibition of topoisomerase (DNA) I (TOP1): DNA damage repair and anticancer therapy. Biomolecules. 5:1652–1670. 2015. View Article : Google Scholar : PubMed/NCBI
32	Rocha JC, Busatto FF, Guecheva TN and Saffi J: Role of nucleotide excision repair proteins in response to DNA damage induced by topoisomerase II inhibitors. Mutat Res Rev Mutat Res. 768:68–77. 2016. View Article : Google Scholar : PubMed/NCBI
33	de Lange T: Shelterin-mediated telomere protection. Ann Rev Genet. 52:223–247. 2018. View Article : Google Scholar : PubMed/NCBI
34	Maiato H and Pereira AJ: Cell division: NuMA bears the load in the spindle. Curr Biol. 27:R765–R767. 2017. View Article : Google Scholar : PubMed/NCBI
35	Gallini S, Carminati M, De Mattia F, Pirovano L, Martini E, Oldani A, Asteriti IA, Guarguaglini G and Mapelli M: NuMA phosphorylation by Aurora-A orchestrates spindle orientation. Curr Biol. 26:458–469. 2016. View Article : Google Scholar : PubMed/NCBI
36	Kotak S and Gonczy P: Mechanisms of spindle positioning: Cortical force generators in the limelight. Curr Opin Cell Biol. 25:741–748. 2013. View Article : Google Scholar : PubMed/NCBI
37	Werner H, Sarfstein R and Bruchim I: Investigational IGF1R inhibitors in early stage clinical trials for cancer therapy. Expert Opin Investig Drugs. 28:1101–1112. 2019. View Article : Google Scholar : PubMed/NCBI
38	Qu X, Wu Z, Dong W, Zhang T, Wang L, Pang Z, Ma W and Du J: Update of IGF-1 receptor inhibitor (ganitumab, dalotuzumab, cixutumumab, teprotumumab and figitumumab) effects on cancer therapy. Oncotarget. 8:29501–29518. 2017. View Article : Google Scholar : PubMed/NCBI
39	Yee D: Anti-insulin-like growth factor therapy in breast cancer. J Mol Endocrinol. 61:T61–T68. 2018. View Article : Google Scholar : PubMed/NCBI
40	Pollak M: The insulin and insulin-like growth factor receptor family in neoplasia: An update. Nat Rev Cancer. 12:159–169. 2012. View Article : Google Scholar : PubMed/NCBI
41	Aleksic T, Gray N, Wu X, Rieunier G, Osher E, Mills J, Verrill C, Bryant RJ, Han C, Hutchinson K, et al: Nuclear IGF1R interacts with regulatory regions of chromatin to promote RNA Polymerase II recruitment and gene expression associated with advanced tumor stage. Cancer Res. 78:3497–3509. 2018.PubMed/NCBI
42	Solomon-Zemler R, Pozniak Y, Geiger T and Werner H: Identification of nucleolar protein NOM1 as a novel nuclear IGF1R-interacting protein. Mol Genet Metab. 126:259–265. 2019. View Article : Google Scholar : PubMed/NCBI
43	Solomon-Zemler R, Sarfstein R and Werner H: Nuclear insulin-like growth factor-1 receptor (IGF1R) displays proliferative and regulatory activities in non-malignant cells. PLoS One. 12:e01851642017. View Article : Google Scholar : PubMed/NCBI
44	Wickramasinghe VO and Venkitaraman AR: RNA processing and genome stability: Cause and consequence. Mol Cell. 61:496–505. 2016. View Article : Google Scholar : PubMed/NCBI
45	Radulescu AE and Cleveland DW: NuMA after 30 years: The matrix revisited. Trends Cell Biol. 20:214–222. 2010. View Article : Google Scholar : PubMed/NCBI
46	Wu J, Xu Z, He D and Lu G: Identification and characterization of novel NuMA isoforms. Biochem Biophys Res Commun. 454:387–392. 2014. View Article : Google Scholar : PubMed/NCBI
47	Panier S and Boulton SJ: Double-strand break repair: 53BP1 comes into focus. Nat Rev Mol Cell Biol. 15:7–18. 2014. View Article : Google Scholar : PubMed/NCBI
48	Serra-Marques A, Houtekamer R, Hintzen D, Canty JT, Yildiz A and Dumont S: The mitotic protein NuMA plays a spindle-independent role in nuclear formation and mechanics. J Cell Biol. 219:e2020042022020. View Article : Google Scholar : PubMed/NCBI