Increased expression of Cyr61 (CCN1) identified in peritoneal metastases from human pancreatic cancer

ArticleinJournal of the American College of Surgeons 200(3):371-7 · April 2005with47 Reads
DOI: 10.1016/j.jamcollsurg.2004.10.005 · Source: PubMed
Identification of extracellular matrix proteins (ECM) associated with tumor cell metastasis may generate targets for future therapy against pancreatic cancer metastases. We hypothesized that comparison of ECM-associated gene expression in primary and metastatic pancreatic tumors would identify ECM proteins associated with pancreatic metastasis. A clinically relevant model of pancreatic cancer was used to generate RNA from primary and metastatic tumors; it was evaluated by microarray analysis with subsequent cluster analysis. Target genes (Cyr61 and integrins alpha(v) and beta(3)) identified by microarray analysis were confirmed by reverse transcription polymerase chain reaction and immunohistochemistry analysis. Peritoneal metastases at sites distant from the primary tumor were present in all animals bearing orthotopic tumors. High-density microarray comparison of gene expression in metastases versus primary pancreatic tumors identified a greater than twofold increase in the expression of Cyr61, a secreted matricellular protein that binds to integrins. Reverse transcription polymerase chain reaction confirmed the microarray results, and immunohistochemistry analysis demonstrated increased Cyr61 protein and persistent alpha(v)beta(3) expression in peritioneal metastases. Additionally, immunohistochemistry demonstrated increased collocalization of Cyr61 and alpha(v) in metastases relative to primary tumor. The ECM protein Cyr61 shows increased expression in metastatic lesions in a clinically relevant model of pancreatic adenocarcinoma. Protein analysis confirms the microarray results and collocalization of Cyr61, and alpha(v) suggests that interaction between Cyr61 and alpha(v)beta(3) promotes formation of peritoneal metastases.
    • "In an orthotopic mouse model of pancreatic cancer, CCN1 showed increased expression in metastatic lesions. This study suggested that interaction between CCN1 and integrins (αvβ3) may promote formation of peritoneal metastases (Holloway et al. 2005 ); yet, its importance in PC was not clearly understood until we uncovered the precise role of CCN1 in PC development (Fig. 2). Our studies demonstrate that when overexpressed in PC and its precursor lesions, CCN1 promotes proliferation, epithelial to mesenchymal transition (EMT), stemness, migration and the growth of xenograft tumors of PC cells (Haque et al. 2011; Leask 2011 ). "
    [Show abstract] [Hide abstract] ABSTRACT: Decades of basic and translational studies have identified the mechanisms by which pancreatic cancer cells use molecular pathways to hijack the normal homeostasis of the pancreas, promoting pancreatic cancer initiation, progression, and metastasis, as well as drug resistance. These molecular pathways were explored to develop targeted therapies to prevent or cure this fatal disease. Regrettably, the studies found that majority of the molecular events that dictate carcinogenic growth in the pancreas are non-actionable (potential non-responder groups of targeted therapy). In this review we discuss exciting discoveries on CCN-siblings that reveal how CCN-family members contribute to the different aspects of the development of pancreatic cancer with special emphasis on therapy.
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    • "It was demonstrated a role in mediating cancer cell invasiveness [44]. Integrin alpha-M was implicated in various adhesive interactions and the expression level was decreased in metastases [45]. It was quantified with four degraded glycosites N696, N58, N1022 and N391 (H/L = 0.58, 0.57, 0.54 and 0.53, respectively). "
    [Show abstract] [Hide abstract] ABSTRACT: Relative quantification of N-glycoproteomes shows great promise for the discovery of candidate biomarkers and therapeutic targets. The traditional protocol for quantitative analysis of glycoproteomes is usually off-line performed, and suffers from long sample preparation time, and the risk of sample loss or contamination due to manual manipulation. In this study, a novel integrated sample preparation platform for quantitative N-glycoproteome analysis was established, with combination of online N-glycopeptide capture by a HILIC column, sample buffer exchange by a N2-assisted HILIC-RPLC interface, deglycosylation by a hydrophilic PNGase F immobilized enzymatic reactor (hIMER) and solid dimethyl labeling on a C18 precolumn. To evaluate the performance of such a platform, two equal aliquots of immunoglobulin G (IgG) digests were sequentially pretreated, followed by MALDI-TOF MS analysis. The signal intensity ratio of heavy/light (H/L) labeled deglycosylated peptides with the equal aliquots was 1.00 (RSD=6.2%, n=3), much better than those obtained by the offline protocol, with H/L ratio as 0.76 (RSD=11.6%, n=3). Additionally, the total on-line sample preparation time was greatly shortened to 160min, much faster than that of offline approach (24h). Furthermore, such an integrated pretreatment platform was successfully applied to analyze the two kinds of hepatocarcinoma ascites syngeneic cell lines with high (Hca-F) and low (Hca-P) lymph node metastasis rates. For H/L labeled Hca-P lysates with the equal aliquots, 99.6% of log2 ratios (H/L) of quantified glycopeptides ranged from -1 to 1, demonstrating high accuracy of the developed sample preparation strategy. By triplicated analysis of glycopeptides and non-glycopeptides of Hca-F and Hca-P lysates, 43 up-regulated and 30 down-regulated (Hca-F/P) N-glycosylation sites, and 11 significantly changed N-glycoproteins were successfully quantified, and most of them were related to tumorigenesis and tumor metastasis. All these results demonstrate the developed integrated N-glycoprotein pretreatment platform is of great power for the accurate, precise and high-throughput analysis of N-glycoproteomes.
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    • "Currently, 24 integrin subtypes have been reported [17, 18], while í µí»¼ví µí»½3 integrin that is overexpressed on tumor cells is one of the most prominent receptors involved in tumor growth, invasiveness, and metastasis [16,19202122. The í µí»¼v integrins and neuropilin-1 are expressed on pancreatic cancer cells, including the panc-1 cell line23242526272829. In addition, í µí»¼ví µí»½3 integrin can be targeted by peptides with a short amino acid sequence containing Arg-Gly-Asp (RGD) [3, 22,303132. "
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