Original contributionSystematic analysis of proteins from different signaling pathways in the tumor center and the invasive front of colorectal cancer
Introduction
The ability of invasion at the tumor front is due to both intrinsic features of the neoplastic cells, such as loss of epithelial differentiation [1], and favorable conditions of the tumor microenvironment [2]. Cells at the invasive front of colorectal carcinomas are considered to have a more aggressive behavior in comparison with more central regions [3]. It could thus be hypothesized that the neoplastic cells at the tumor front adapt their phenotype to satisfy the needs of their new role, which differs from the role of the neoplastic cells located in the main tumor body.
The expression and functional impact of proteins from various signaling pathways have been reported to be different between the tumor center and the tumor front in colorectal cancer (CRC) [3], [4], [5]. Indeed, the interaction between malignant epithelium and stromal elements, also called epithelial-mesenchymal transition, occurring at the invasive front is recognized as a crucial step in CRC carcinogenesis [6].
In CRC, TNM stage and other established morphological features provide the standard criterion for identifying patients with different risks of a specific outcome. However, the identification of reproducible and well-characterized predictive and prognostic biomarkers would help in defining subgroups in CRC.
The aim of the present study was to investigate potential differences in protein marker expression on 220 well-characterized patients with CRC with the ability to differentiate between the tumor center and the tumor front to identify prognostic factors for CRCs. Established and promising prognostic proteins selected to represent various signaling pathways including the wingless-INT signaling pathway (Wnt) (β-catenin, E-cadherin), extracellular signal regulated kinase/mitogen activated protein kinase (ERK/MAPK) (EGFR, pERK, Raf-1 kinase inhibitor protein [RKIP]), AKT (pAKT), transforming growth factor β (TGF-β) (pSMAD2), as well as proteins involved in cell cycle regulation, apoptosis, or proliferation (p21, p16, BCL2, Ki-67, apoptosis protease activating factor-1 [APAF-1], MST1) and in angiogenic or metastatic processes (RHAMM, vascular endothelial growth factor [VEGF], EphB2, matrix metalloproteinase [MMP7], Laminin5γ2, MUC1, CDX2) were evaluated using a multiple-punch tissue microarray.
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Patients and clinicopathological features
Two hundred twenty nonconsecutive patients treated at the 4th Department of Surgery, University of Athens Medical School were entered into the study. Paraffin blocks were selected from the archives of the 2nd Department of Pathology, University of Athens Medical School (Attikon University Hospital), Greece. All selected cases had at least 1.2 cm tumor tissue thickness in at least 2 paraffin blocks to be included to the study.
Patients were treated between 2004 and 2006. All histomorphological
Immunohistochemistry
The number of evaluable tissue punches for each protein marker was as follows: EGFR, 791; RHAMM, 783; β-catenin, 799; p16, 788; p21, 800; Ki-67, 786; pAKT, 779; BCL2, 778; VEGF, 775; APAF-1, 766; MUC1, 792; pERK, 776; EphB2, 779; E-cadherin, 787; RKIP, 767; MMP-7, 781; pSMAD, 352; CDX2, 782; Laminin5γ2, 783; and MST1, 776. Immunohistochemical results are summarized in Table 2, Table 3, Table 4.
Differences in mean protein expression between the invasive tumor front and the tumor center
The mean protein expression between the tumor front and the tumor center varied significantly for
Discussion
Our results demonstrate that VEGF, RKIP, and Laminin5γ2 were the most relevant proteins, with the multimarker phenotypes positive/positive/negative and negative/negative/positive being most discriminating between the tumor center and the tumor front in CRC.
We found a differential VEGF expression between the tumor center and the tumor front in CRC, with loss of VEGF expression at the invasive tumor front. Vascular endothelial growth factor is a potent proangiogenic factor whose downstream
References (32)
- et al.
Heterogeneous expression of cyclooxygenase-2 and inducible nitric oxide synthase within colorectal tumors: correlation with tumor angiogenesis
Dig Liver Dis
(2010) - et al.
Nuclear overexpression of the oncoprotein β-catenin in colorectal cancer is localized predominantly at the invasion front
Pathol Res Pract
(1998) Vascular endothelial growth factor (VEGF) signalling in tumor progression
Crit Rev Oncol Hematol
(2007)- et al.
Raf kinase inhibitory protein regulates Raf-1 but not B-Raf kinase activation
J Biol Chem
(2005) - et al.
Role of the mitogen-activated protein kinase and phosphoinositide 3 kinase/AKT pathways downstream molecules, phosphorylated extracellular signal-regulated kinase, and phosphorylated AKT in colorectal cancer: a tissue microarray-based approach
Hum Pathol
(2006) - et al.
The expression of E-cadherin and catenins in sporadic colorectal carcinoma
Hum Pathol
(2001) - et al.
Genetic manipulation of E-cadherin expression by epithelial tumor cells reveals an invasion suppressor role
Cell
(1991) - et al.
Tumour “budding” as an index to estimate the potential of aggressiveness in rectal cancer
Histopathology
(2002) - et al.
The microenvironment of the tumour-host interface
Nature
(2001) - et al.
MUC1 and nuclear beta-catenin are coexpressed at the invasion front of colorectal carcinomas and are both correlated with tumor prognosis
Clin Cancer Res
(2004)
Down-regulation of the homeodomain factor Cdx2 in colorectal cancer by collagen type I: an active role for the tumor environment in malignant tumor progression
Cancer Res
VEGF as a predictive marker of rectal tumor response to preoperative radiotherapy
Cancer
Microvessel quantification and its possible relation with liver metastasis in colorectal cancer
Cancer
Angiogenesis as an unfavourable prognostic factor in human colorectal carcinoma
Cancer
Angiogenesis in health and disease
Nat Med
Expression of vascular endothelial growth factor and its receptor, KDR, correlates with vascularity, metastasis and proliferation of human colon cancer
Cancer Res
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