Interaction between GRP78 and IGFBP-3 Affects Tumourigenesis and Prognosis in Breast Cancer Patients
Abstract
:Simple Summary
Abstract
1. Introduction
2. Results
2.1. Correlation between the Expression of GRP78 and IGFBP-3 in Breast Cell Lines and Clinical Samples
2.2. The Absence of GRP78 Modifies the Actions of IGFBP-3 In Vitro
2.3. IGFBP-3 Subcellular Localisation Is Regulated by GRP78
2.4. Loss of GRP78 Predicts a Poor Prognosis in Breast Cancer Patients
3. Discussion
4. Materials and Methods
4.1. Cell Culture
4.2. Dosing with IGFBP-3 and siRNA Transfection
4.3. Transwell Invasion Assay
4.4. Western Blotting
4.5. Immunoprecipitation
4.6. Immunofluorescence Staining and Confocal Imaging
4.7. Cell Surface Biotinylation and Avidin Pulldown
4.8. Human Tissue Samples
4.9. Immunohistochemistry
4.10. Molecular Taxonomy of Breast Cancer International Consortium (METABRIC) Dataset Analysis
4.11. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
- American Institute for Cancer Research. Cancer Statistics. Available online: https://www.aicr.org/ (accessed on 1 October 2020).
- Cancer Research UK. Cancer Statistics. Available online: https://www.cancerresearchuk.org/health-professional/cancer-statistics/statistics-by-cancer-type/breast-cancer (accessed on 1 October 2020).
- Mallini, P.; Lennard, T.; Kirby, J.; Meeson, A. Epithelial-to-mesenchymal transition: What is the impact on breast cancer stem cells and drug resistance. Cancer Treat. Rev. 2014, 40, 341–348. [Google Scholar] [CrossRef] [PubMed]
- Perks, C.M.; Holly, J.M. IGF binding proteins (IGFBPs) and regulation of breast cancer biology. J. Mammary Gland Biol. Neoplasia 2008, 13, 455–469. [Google Scholar] [CrossRef] [PubMed]
- Christopoulos, P.F.; Msaouel, P.; Koutsilieris, M. The role of the insulin-like growth factor-1 system in breast cancer. Mol. Cancer 2015, 14, 43. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Scully, T.; Scott, C.D.; Firth, S.M.; Pintar, J.E.; Twigg, S.M.; Baxter, R.C. Contrasting effects of IGF binding protein-3 expression in mammary tumor cells and the tumor microenvironment. Exp. Cell Res. 2019, 374, 38–45. [Google Scholar] [CrossRef] [PubMed]
- Burrows, C.; Holly, J.M.; Laurence, N.J.; Vernon, E.G.; Carter, J.V.; Clark, M.A.; McIntosh, J.; McCaig, C.; Winters, Z.E.; Perks, C.M. Insulin-like growth factor binding protein 3 has opposing actions on malignant and nonmalignant breast epithelial cells that are each reversible and dependent upon cholesterol-stabilized integrin receptor complexes. Endocrinology 2006, 147, 3484–3500. [Google Scholar] [CrossRef] [Green Version]
- Kim, J.H.; Cho, Y.H.; Park, Y.L.; Sohn, J.H.; Kim, H.S. Prognostic significance of insulin growth factor-I receptor and insulin growth factor binding protein-3 expression in primary breast cancer. Oncol. Rep. 2010, 23, 989–995. [Google Scholar] [CrossRef]
- Rocha, R.L.; Hilsenbeck, S.G.; Jackson, J.G.; Lee, A.V.; Figueroa, J.A.; Yee, D. Correlation of insulin-like growth factor-binding protein-3 messenger RNA with protein expression in primary breast cancer tissues: Detection of higher levels in tumors with poor prognostic features. J. Natl. Cancer Inst. 1996, 88, 601–606. [Google Scholar] [CrossRef] [Green Version]
- Zielinska, H.A.; Bahl, A.; Holly, J.M.; Perks, C.M. Epithelial-to-mesenchymal transition in breast cancer: A role for insulin-like growth factor I and insulin-like growth factor-binding protein 3? Breast Cancer (Dove Med. Press) 2015, 7, 9–19. [Google Scholar] [CrossRef] [Green Version]
- Julovi, S.M.; Martin, J.L.; Baxter, R.C. Nuclear Insulin-Like Growth Factor Binding Protein-3 As a Biomarker in Triple-Negative Breast Cancer Xenograft Tumors: Effect of Targeted Therapy and Comparison with Chemotherapy. Front. Endocrinol. 2018, 9, 120. [Google Scholar] [CrossRef] [Green Version]
- Grkovic, S.; O’Reilly, V.C.; Han, S.; Hong, M.; Baxter, R.C.; Firth, S.M. IGFBP-3 binds GRP78, stimulates autophagy and promotes the survival of breast cancer cells exposed to adverse microenvironments. Oncogene 2013, 32, 2412–2420. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Li, C.; Harada, A.; Oh, Y. IGFBP-3 sensitizes antiestrogen-resistant breast cancer cells through interaction with GRP78. Cancer Lett. 2012, 325, 200–206. [Google Scholar] [CrossRef] [PubMed]
- Tsai, Y.-L.; Zhang, Y.; Tseng, C.-C.; Stanciauskas, R.; Pinaud, F.; Lee, A.S. Characterization and mechanism of stress-induced translocation of 78-kilodalton glucose-regulated protein (GRP78) to the cell surface. J. Biol. Chem. 2015, 290, 8049–8064. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- McCaig, C.; Perks, C.M.; Holly, J.M. Intrinsic actions of IGFBP-3 and IGFBP-5 on Hs578T breast cancer epithelial cells: Inhibition or accentuation of attachment and survival is dependent upon the presence of fibronectin. J. Cell Sci. 2002, 115, 4293–4303. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Ni, M.; Zhang, Y.; Lee, A.S. Beyond the endoplasmic reticulum: Atypical GRP78 in cell viability, signalling and therapeutic targeting. Biochem. J. 2011, 434, 181–188. [Google Scholar] [CrossRef]
- Quinones, Q.J.; de Ridder, G.G.; Pizzo, S.V. GRP78: A chaperone with diverse roles beyond the endoplasmic reticulum. Histol. Histopathol. 2008, 23, 1409–1416. [Google Scholar] [CrossRef]
- Baxter, R.C. Nuclear actions of insulin-like growth factor binding protein-3. Gene 2015, 569, 7–13. [Google Scholar] [CrossRef] [Green Version]
- Gonzalez-Gronow, M.; Selim, M.A.; Papalas, J.; Pizzo, S.V. GRP78: A multifunctional receptor on the cell surface. Antioxid. Redox Signal. 2009, 11, 2299–2306. [Google Scholar] [CrossRef]
- Casas, C. GRP78 at the Centre of the Stage in Cancer and Neuroprotection. Front. Neurosci. 2017, 11, 177. [Google Scholar] [CrossRef]
- Li, Z.; Zhang, L.; Zhao, Y.; Li, H.; Xiao, H.; Fu, R.; Zhao, C.; Wu, H.; Li, Z. Cell-surface GRP78 facilitates colorectal cancer cell migration and invasion. Int. J. Biochem. Cell Biol. 2013, 45, 987–994. [Google Scholar] [CrossRef]
- Zhang, X.X.; Li, H.D.; Zhao, S.; Zhao, L.; Song, H.J.; Wang, G.; Guo, Q.J.; Luan, Z.D.; Su, R.J. The cell surface GRP78 facilitates the invasion of hepatocellular carcinoma cells. BioMed Res. Int. 2013, 2013, 917296. [Google Scholar] [CrossRef] [Green Version]
- Rocha, R.L.; Hilsenbeck, S.G.; Jackson, J.G.; VanDenBerg, C.L.; Weng, C.; Lee, A.V.; Yee, D. Insulin-like growth factor binding protein-3 and insulin receptor substrate-1 in breast cancer: Correlation with clinical parameters and disease-free survival. Clin. Cancer Res. 1997, 3, 103–109. [Google Scholar] [PubMed]
- Yu, H.; Levesque, M.A.; Khosravi, M.J.; Papanastasiou-Diamandi, A.; Clark, G.M.; Diamandis, E.P. Associations between insulin-like growth factors and their binding proteins and other prognostic indicators in breast cancer. Br. J. Cancer 1996, 74, 1242–1247. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Vestey, S.B.; Perks, C.M.; Sen, C.; Calder, C.J.; Holly, J.M.; Winters, Z.E. Immunohistochemical expression of insulin-like growth factor binding protein-3 in invasive breast cancers and ductal carcinoma in situ: Implications for clinicopathology and patient outcome. Breast Cancer Res. 2005, 7, R119–R129. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Song, G.; Liu, K.; Zhu, X.; Yang, X.; Shen, Y.; Wang, W.; Shi, G.; Li, Q.; Duan, Y.; Zhao, Y.; et al. The low IGFBP-3 level is associated with esophageal cancer patients: A meta-analysis. World J. Surg. Oncol. 2016, 14, 307. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Torng, P.L.; Lee, Y.C.; Huang, C.Y.; Ye, J.H.; Lin, Y.S.; Chu, Y.W.; Huang, S.C.; Cohen, P.; Wu, C.W.; Lin, C.T. Insulin-like growth factor binding protein-3 (IGFBP-3) acts as an invasion-metastasis suppressor in ovarian endometrioid carcinoma. Oncogene 2008, 27, 2137–2147. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Perks, C.M.; Burrows, C.; Holly, J.M. Intrinsic, Pro-Apoptotic Effects of IGFBP-3 on Breast Cancer Cells are Reversible: Involvement of PKA, Rho, and Ceramide. Front. Endocrinol. 2011, 2, 13. [Google Scholar] [CrossRef] [Green Version]
- Zhang, J.; Jiang, Y.; Jia, Z.; Li, Q.; Gong, W.; Wang, L.; Wei, D.; Yao, J.; Fang, S.; Xie, K. Association of elevated GRP78 expression with increased lymph node metastasis and poor prognosis in patients with gastric cancer. Clin. Exp. Metastasis 2006, 23, 401–410. [Google Scholar] [CrossRef]
- Niu, Z.; Wang, M.; Zhou, L.; Yao, L.; Liao, Q.; Zhao, Y. Elevated GRP78 expression is associated with poor prognosis in patients with pancreatic cancer. Sci. Rep. 2015, 5, 16067. [Google Scholar] [CrossRef] [Green Version]
- Ren, P.; Chen, C.; Yue, J.; Zhang, J.; Yu, Z. High expression of glucose-regulated protein 78 (GRP78) is associated with metastasis and poor prognosis in patients with esophageal squamous cell carcinoma. OncoTargets Ther. 2017, 10, 617–625. [Google Scholar] [CrossRef]
- Yu, D.; Luo, Y.; Liu, G. Immunohistochemical GRP78 as a tumor biomarker may predict poor prognosis in patients with non-small cell lung cancers: A retrospective study. Biomed. Res. 2016, 27, 948–953. [Google Scholar]
- Fernandez, P.M.; Tabbara, S.O.; Jacobs, L.K.; Manning, F.C.; Tsangaris, T.N.; Schwartz, A.M.; Kennedy, K.A.; Patierno, S.R. Overexpression of the glucose-regulated stress gene GRP78 in malignant but not benign human breast lesions. Breast Cancer Res. Treat. 2000, 59, 15–26. [Google Scholar] [CrossRef] [PubMed]
- Quach, H.; North, D.; Freddi, S.; Tan, S.; Straszkowski, L.; Nandurkar, H. High GRP78 (78-kDa Glucose-Regulated Protein) Expression Predicts for a Favorable Clinical Outcome in Patients with Multiple Myeloma and May be a Potentially Useful Therapeutic Target in the Treatment of Multiple Myeloma. Blood 2015, 126, 4206. [Google Scholar] [CrossRef]
- Yerushalmi, R.; Raiter, A.; Nalbandyan, K.; Hardy, B. Cell surface GRP78: A potential marker of good prognosis and response to chemotherapy in breast cancer. Oncol. Lett. 2015, 10, 2149–2155. [Google Scholar] [CrossRef] [PubMed]
- Al-Keilan, M.; Almomani, B.; Alqudah, M.; Alrjoub, M.; Alzoubi, H.; Shhabat, B. Immunohistochemical expression of GRP78 in relation to angiogenesis markers VEGF-a and CD31 and other histopathological parameters in NSCLC. J. Clin. Oncol. 2019, 37, e20500. [Google Scholar] [CrossRef]
- Zeng, L.; Biernacka, K.M.; Holly, J.M.; Jarrett, C.; Morrison, A.A.; Morgan, A.; Winters, Z.E.; Foulstone, E.J.; Shield, J.P.; Perks, C.M. Hyperglycaemia confers resistance to chemotherapy on breast cancer cells: The role of fatty acid synthase. Endocr. Relat. Cancer 2010, 17, 539–551. [Google Scholar] [CrossRef] [Green Version]
- Zeng, L.; Holly, J.M.; Perks, C.M. Effects of physiological levels of the green tea extract epigallocatechin-3-gallate on breast cancer cells. Front. Endocrinol. 2014, 5, 61. [Google Scholar] [CrossRef] [Green Version]
- Lee, E.; Nichols, P.; Spicer, D.; Groshen, S.; Yu, M.C.; Lee, A.S. GRP78 as a novel predictor of responsiveness to chemotherapy in breast cancer. Cancer Res. 2006, 66, 7849–7853. [Google Scholar] [CrossRef] [Green Version]
- Lee, E.; Nichols, P.; Groshen, S.; Spicer, D.; Lee, A.S. GRP78 as potential predictor for breast cancer response to adjuvant taxane therapy. Int. J. Cancer 2011, 128, 726–731. [Google Scholar] [CrossRef] [Green Version]
Total | ||
---|---|---|
Age | <50 | 11 |
50–70 | 41 | |
>70 | 17 | |
Tumour size | ≤2 cm | 34 |
2 cm–≤ 5 cm | 33 | |
>5 cm | 2 | |
Grade | 1 | 2 |
2 | 30 | |
3 | 37 | |
ER | Pos | 23 |
Neg | 46 | |
PR | Pos | 18 |
Neg | 51 | |
Her2+ | Pos | 25 |
Neg | 44 | |
TN | Yes | 30 |
no | 39 | |
Histology | Ductal | 59 |
other | 10 | |
Lymph involvment | Yes | 25 |
No | 44 |
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. |
© 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
Share and Cite
Zielinska, H.A.; Daly, C.S.; Alghamdi, A.; Bahl, A.; Sohail, M.; White, P.; Dean, S.R.; Holly, J.M.P.; Perks, C.M. Interaction between GRP78 and IGFBP-3 Affects Tumourigenesis and Prognosis in Breast Cancer Patients. Cancers 2020, 12, 3821. https://doi.org/10.3390/cancers12123821
Zielinska HA, Daly CS, Alghamdi A, Bahl A, Sohail M, White P, Dean SR, Holly JMP, Perks CM. Interaction between GRP78 and IGFBP-3 Affects Tumourigenesis and Prognosis in Breast Cancer Patients. Cancers. 2020; 12(12):3821. https://doi.org/10.3390/cancers12123821
Chicago/Turabian StyleZielinska, Hanna A., Carl S. Daly, Ahmad Alghamdi, Amit Bahl, Muhammed Sohail, Paul White, Sarah R. Dean, Jeff M. P. Holly, and Claire M. Perks. 2020. "Interaction between GRP78 and IGFBP-3 Affects Tumourigenesis and Prognosis in Breast Cancer Patients" Cancers 12, no. 12: 3821. https://doi.org/10.3390/cancers12123821