Abstract
Aim and background
Epidermal growth factor receptor (EGFR) is a receptor tyrosine kinase involved in many important aspects of cell biology that are related to tumorigenesis. There are opposite evidences of the role of EGFR in renal cancer and the outcome of EGFR-targeted therapies, suggesting the complexity of EGFR signaling pathways. In vitro, osteopontin (OPN) and nuclear factor kappa B (NF-κB) are thought to be involved in specific ligand-independent EGFR activation that could have a role in resistance to EGFR mAb therapy. Aim of this study was to analyze the relationship between EGFR and OPN at the protein and mRNA level, as well as their relation to NF-κB in clear cell renal cell carcinoma (CCRCC).
Materials and methods
Expression of EGFR, OPN, and p65 NF-κB protein was analyzed using immunohistochemistry and compared mutually in 88 CCRCC samples. Expression of EGFR and OPN mRNAs was analyzed using quantitative Real-time PCR in 22 CCRCC samples and compared mutually and with NF-κB protein expression.
Results
Epidermal growth factor receptor mRNA level was higher in CCRCC samples in comparison with normal renal tissue (p = 0.012) and was associated with high OPN mRNA level, and with NF-κB activation (p < 0.001 and p = 0.045, respectively). Immunohistochemical staining showed the inverse association; high EGFR protein expression was related with low OPN and NF-κB protein expression (p < 0.001 and p = 0.047, respectively).
Conclusion
Epidermal growth factor receptor gene is upregulated in CRCC and associated with OPN gene expression and NF-kB signaling. The inverse relation between OPN and EGFR at the protein level could probably reflect dynamic changes that EGFR undergoes following activation.
Similar content being viewed by others
References
Wells A (1999) EGF receptor. Int J Biochem Cell Biol 31:637–643
Prenzel N, Fischer OM, Streit S, Hart S, Ullrich A (2001) The epidermal growth factor receptor family as a central element for cellular signal transduction and diversification. Endocr Relat Cancer 8:11–31
Lu Z, Jiang G, Blume-Jensen P, Hunter T (2001) Epidermal growth factor-induced tumor cell invasion and metastasis initiated by dephosphorylation and downregulation of focal adhesion kinase. Mol Cell Biol 21:4016–4031
Goel S, Hidalgo M, Perez-Soler R (2007) EGFR inhibitor-mediated apoptosis in solid tumors. J Exp Ther Oncol 6:305–320
Wheeler DL, Dunn EF, Harari PM (2010) Understanding resistance to EGFR inhibitors-impact on future treatment strategies. Nat Rev Clin Oncol 7:493–507
Normanno N, De Luca A, Bianco C, Strizzi L, Mancino M, Maiello MR, Carotenuto A, De Feo G, Caponigro F, Salomon DS (2006) Epidermal growth factor receptor (EGFR) signaling in cancer. Gene 366:2–16
Streuli CH, Akhtar N (2009) Signal co-operation between integrins and other receptor systems. Biochem J 418:491–506
Cabodi S, del Pilar Camacho-Leal M, Di Stefano P, Defilippi P (2010) Integrin signalling adaptors: not only figurants in the cancer story. Nat Rev Cancer 10:858–870
Rittling SR, Chambers AF (2008) Role of osteopontin in tumour progression. Br J Cancer 90:1877–1881
Wai PY, Kuo PC (2008) Osteopontin: regulation in tumor metastasis. Cancer Metastasis Rev 27:103–118
Coppola D, Szabo M, Boulware D, Muraca P, Alsarraj M, Chambers AF, Yeatman TJ (2004) Correlation of osteopontin protein expression and pathological stage across a wide variety of tumor histologies. Clin Cancer Res 10:184–190
Standal T, Borset M, Sundan A (2004) Role of osteopontin in adhesion, migration, cell survival and bone remodeling. Exp Oncol 26:179–184
El-Tanani MK, Campbell FC, Kurisetty V, Jin D, McCann M, Rudland PS (2006) The regulation and role of osteopontin in malignant transformation and cancer. Cytokine Growth Factor Rev 6:463–474
Tuck AB, Hota C, Wilson SM, Chambers AF (2003) Osteopontin-induced migration of human mammary epithelial cells involves activation of EGF receptor and multiple signal transduction pathways. Oncogene 22:1198–1205
Philip S, Bulbule A, Kundu GC (2001) Osteopontin stimulates tumor growth and activation of promatrix metalloproteinase-2 through nuclear factor-kappa B-mediated induction of membrane type 1 matrix metalloproteinase in murine melanoma cells. J Biol Chem 276:44926–44935
Matusan K, Dordevic G, Stipic D, Mozetic V, Lucin K (2006) Osteopontin expression correlates with prognostic variables and survival in clear cell renal-cell carcinoma. J Surg Oncol 4:325–331
Matušan-Ilijaš K, Damante G, Fabbro D, Đorđević G, Hadžisejdić I, Grahovac M, Marić I, Španjol J, Grahovac B, Jonjić N, Lučin K (2011) Osteopontin expression correlates with nuclear factor-kappa B activation and apoptosis downregulation in clear cell renal cell carcinoma. Pathol Res Pract 207:104–110
Zhang G, He B, Weber GF (2003) Growth factor signaling induces metastasis genes in transformed cells: molecular connection between Akt kinase and osteopontin in breast cancer. Mol Cell Biol 23:6507–6519
Malyankar UM, Almeida M, Johnson RJ, Pichler RH, Giachelli CM (1997) Osteopontin regulation in cultured rat renal epithelial cells. Kidney Int 51:1766–1773
Zhang G, Zhao ZQ, Wang HD, Hao B (2004) Enhancement of osteopontin expression in HepG2 cells by epidermal growth factor via phosphatidylinositol 3-kinase signaling pathway. World J Gastroenterol 10:205–208
Chakalaparampil I, Peri A, Nemir M, Mckee MD, Lin PH, Mukherjee BB, Mukherjee AB (1996) Cells in vivo and in vitro from osteopetrotic mice homozygous for c-src disruption show suppression of synthesis of osteopontin, a multifunctional extracellular matrix protein. Oncogene 12:1457–1467
Ivaska J, Heino J (2010) Interplay between cell adhesion and growth factor receptors: from the plasma membrane to the endosomes. Cell Tissue Res 339:111–120
Angelluci A, Festuccia C, Gravina GL, Muzi P, Bonghi L, Vicentini C, Bologna M (2004) Osteopontin enhances the cell proliferation induced by the epidermal growth factor in human prostate cancer cells. Prostate 59:157–166
Langner C, Ratschek M, Rehak P, Schips L, Zigeuner R (2004) Are heterogenous results of EGFR immunoreactivity in renal cell carcinoma related to non-standardised criteria for staining evaluation? J Clin Pathol 57:773–775
Kallio JP, Hirvikoski P, Helin H, Kellokumpu-Lehtinen P, Luukkaala T, Tammela TL, Martikainen PM (2003) Membranous location of EGFR immunostaining is associated with good prognosis in renal cell carcinoma. Br J Cancer 89:1266–1269
Pu YS, Huang CY, Kuo YZ, Kang WY, Liu GY, Huang AM, Yu HJ, Lai MK, Huang SP, Wu WJ, Chiou SJ, Hour TC (2009) Characterization of membranous and cytoplasmic EGFR expression in human normal renal cortex and renal cell carcinoma. J Biomed Sci 16:82
Merseburger AS, Hennenlotter J, Simon P, Kruck S, Koch E, Horstmann M, Kuehs U, Küfer R, Stenzl A, Kuczyk MA (2005) Membranous expression and prognostic implications of epidermal growth factor receptor protein in human renal cell cancer. Anticancer Res 3:1901–1907
Piazzi G, Paterini P, Ceccarelli C, Pantaleo MA, Biasco G (2006) Molecular determination of epidermal growth factor receptor in normal and neoplastic colorectal mucosa. Br J Cancer 95:1525–1528
Keese M, Magdeburg RJ, Herzog T, Hasenberg T, Offterdinger M, Pepperkok R, Sturm JW, Bastiaens PI (2005) Imaging epidermal growth factor receptor phosphorylation in human colorectal cancer cells and human tissues. J Biol Chem 280:27826–27831
Nannini M, Pantaleo MA, Paterini P, Piazzi G, Ceccarelli C, La Rovere S, Maleddu A, Biasco G (2011) Molecular detection of epidermal growth factor receptor in colorectal cancer: does it still make sense? Colorectal Dis 5:542–548. doi:10.1111/j.1463-1318.2010.02212.x
Dei Tos AP, Ellis I (2005) Assessing epidermal growth factor receptor expression in tumours: what is the value of current test methods? Eur J Cancer 41:1383–1392
Bukowski RM, Kabbinavar FF, Figlin RA, Flaherty K, Srinivas S, Vaishampayan U, Drabkin HA, Dutcher J, Ryba S, Xia Q, Scappaticci FA, McDermott D (2007) Randomized phase II study of erlotinib combined with bevacizumab compared with bevacizumab alone in metastatic renal cell cancer. J Clin Oncol. 25(29):4536–4541
Jermann M, Stahel RA, Salzberg M, Cerny T, Joerger M, Gillessen S, Morant R, Egli F, Rhyner K, Bauer JA, Pless M (2006) A phase II, open-label study of gefitinib (IRESSA) in patients with locally advanced, metastatic, or relapsed renal-cell carcinoma. Cancer Chemother Pharmacol 57:533–539
Hainsworth JD, Sosman JA, Spigel DR, Edwards DL, Baughman C, Greco A (2005) Treatment of metastatic renal cell carcinoma with a combination of bevacizumab and erlotinib. J Clin Oncol 23:7889–7896
Ravaud A, Hawkins R, Gardner JP, von der Maase H, Zantl N, Harper P, Rolland F, Audhuy B, Machiels JP, Pétavy F, Gore M, Schöffski P, El-Hariry I (2008) Lapatinib versus hormone therapy in patients with advanced renal cell carcinoma: a randomized phase III clinical trial. J Clin Oncol 26:2285–2291
Jimeno A, Rubio-Viqueira B, Amador ML, Oppenheimer D, Bouraoud N, Kulesza P, Sebastiani V, Maitra A, Hidalgo M (2005) Epidermal growth factor receptor dynamics influences response to epidermal growth factor receptor targeted agents. Cancer Res 65:3003–3010
Acknowledgments
This work was supported by the Ministry of Science, Education, and Sports of the Republic of Croatia (grant 062-0620095-0078), Alpe-Adria Research Grant and MIUR grant, Italy and TransMedRi GA No. 256686. We thank Mrs. Tanja Kovačević and Mr. Ozren Štanfel for their excellent technical support.
Conflict of interest
None.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Matušan-Ilijaš, K., Damante, G., Fabbro, D. et al. EGFR expression is linked to osteopontin and Nf-κB signaling in clear cell renal cell carcinoma. Clin Transl Oncol 15, 65–71 (2013). https://doi.org/10.1007/s12094-012-0889-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12094-012-0889-9