Intra-tumoural vessel area estimated by expression of epidermal growth factor-like domain 7 and microRNA-126 in primary tumours and metastases of patients with colorectal cancer: a descriptive study

Journal of Translational Medicine

Flemming Brandt Sørensen¹  Anders Jakobsen¹  Boye Schnack Nielsen²  Torben Frøstrup Hansen¹ 

[1] Institute of Regional Health Research, University of Southern Denmark, Region of Southern Denmark, Denmark;Bioneer A/S, Hørsholm, Denmark

关键词: microRNA-126;    Metastases;    Epidermal growth factor like domain 7;    Colorectal neoplasms;    Angiogenesis;   

Others  :  1146205
DOI  :  10.1186/s12967-014-0359-y

 received in 2014-06-17, accepted in 2014-12-10,  发布年份 2015

【 摘 要 】

Background

Understanding the biological properties of potential drug targets are important. This is especially true for anti-angiogenic therapies in the search for potential biomarkers. The aim of the present descriptive study was to analyse the intra-tumoural expressions of epidermal growth factor-like domain 7 (EGFL7) and microRNA-126 (miRNA-126) in primary tumours from patients with stage II-IV colorectal cancer (CRC) and in paired samples of primary tumours, regional lymph node metastases and distant metastases.

Methods

A total of 126 patients were included. Analyses were performed on resections of primary tumours, regional lymph node metastases, and large needle biopsies from distant metastases. EGFL7 was analysed by immunohistochemistry (IHC) and miRNA-126 by in situ hybridization (ISH). Both biomarkers were quantified by image guided analyses to determine the relative fraction estimates of vessel areas (VA).

Results

The intra-tumoural EGFL7 VA was significantly higher in primary tumours from patients with recurrent disease than in patients without relapse in both stage II and III, p = 0.019 and p = 0.001, respectively. The EGFL7 VA was significantly higher and the miRNA-126 VA significantly lower in regional lymph node metastases compared to primary tumours, p = 0.01 and p < 10⁻⁶, respectively. Furthermore, the miRNA-126 VA in liver metastases was significantly lower than in the primary tumours, p = 0.02.

Conclusion

The intra-tumoural expression of EGFL7 in early stages of CRC may influence the risk of post-surgical recurrence. Differential expression of miRNA-126 seems more pronounced in disseminated disease, which supports its role as a regulator in the metastatic process.

【 授权许可】

   
2015 Hansen et al.; licensee BioMed Central.

【 图 表 】

Figure 1.

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【 参考文献 】

• [1]Cao Y, Tan A, Gao F, Liu L, Liao C, Mo Z: A meta-analysis of randomized controlled trials comparing chemotherapy plus bevacizumab with chemotherapy alone in metastatic colorectal cancer. Int J Colorectal Dis 2009, 24:677-85.
• [2]Wagner AD, Arnold D, Grothey AA, Haerting J, Unverzagt S: Anti-angiogenic therapies for metastatic colorectal cancer. Cochrane Database Syst Rev 2009, 3:CD005392. doi:10.1002/14651858.CD005392
• [3]Ferrara N, Gerber HP, LeCouter J: The biology of VEGF and its receptors. Nat Med 2003, 9:669-76.
• [4]Folkman J: What is the evidence that tumors are angiogenesis dependent? J Natl Cancer Inst 1990, 82:4-6.
• [5]Fitch MJ, Campagnolo L, Kuhnert F, Stuhlmann H: Egfl7, a novel epidermal growth factor-domain gene expressed in endothelial cells. Dev Dyn 2004, 230:316-24.
• [6]Parker LH, Schmidt M, Jin SW, Gray AM, Beis D, Pham T, Frantz G, Palmieri S, Hillan K, Stainier DY, De Sauvage FJ, Ye W: The endothelial-cell-derived secreted factor Egfl7 regulates vascular tube formation. Nature 2004, 428:754-8.
• [7]Soncin F, Mattot V, Lionneton F, Spruyt N, Lepretre F, Begue A, Stehelin D: VE-statin, an endothelial repressor of smooth muscle cell migration. EMBO J 2003, 22:5700-11.
• [8]Campagnolo L, Leahy A, Chitnis S, Koschnick S, Fitch MJ, Fallon JT, Loskutoff D, Taubman MB, Stuhlmann H: EGFL7 is a chemoattractant for endothelial cells and is up-regulated in angiogenesis and arterial injury. Am J Pathol 2005, 167:275-84.
• [9]Schmidt M, Paes K, De MA, Smyczek T, Yang S, Gray A, French D, Kasman I, Klumperman J, Rice DS, Ye W: EGFL7 regulates the collective migration of endothelial cells by restricting their spatial distribution. Development 2007, 134:2913-23.
• [10]Delfortrie S, Pinte S, Mattot V, Samson C, Villain G, Caetano B, Lauridant-Philippin G, Baranzelli MC, Bonneterre J, Trottein F, Faveeuw C, Soncin F: Egfl7 promotes tumor escape from immunity by repressing endothelial cell activation. Cancer Res 2011, 71:7176-86.
• [11]Diaz R, Silva J, Garcia JM, Lorenzo Y, Garcia V, Pena C, Rodriguez R, Munoz C, Garcia F, Bonilla F, Dominguez G: Deregulated expression of miR-106a predicts survival in human colon cancer patients. Genes Chromosomes Cancer 2008, 47:794-802.
• [12]Wu F, Yang LY, Li YF, Ou DP, Chen DP, Fan C: Novel role for epidermal growth factor-like domain 7 in metastasis of human hepatocellular carcinoma. Hepatology 2009, 50:1839-50.
• [13]Li JJ, Yang XM, Wang SH, Tang QL: Prognostic role of epidermal growth factor-like domain 7 protein expression in laryngeal squamous cell carcinoma. J Laryngol Otol 2011, 125:1152-7.
• [14]Hansen TF, Christensen R, Andersen RF, Sorensen FB, Johnsson A, Jakobsen A: MicroRNA-126 and epidermal growth factor-like domain 7-an angiogenic couple of importance in metastatic colorectal cancer. Results from the Nordic ACT trial. Br J Cancer 2013, 109:1243-51.
• [15]Wienholds E, Kloosterman WP, Miska E, varez-Saavedra E, Berezikov E, de Bruijn E , Horvitz HR, Kauppinen S, Plasterk RH: MicroRNA expression in zebrafish embryonic development. Science 2005, 309:310-1.
• [16]Zhu N, Zhang D, Xie H, Zhou Z, Chen H, Hu T, Bai Y, Shen Y, Yuan W, Jing Q, Qin Y: Endothelial-specific intron-derived miR-126 is down-regulated in human breast cancer and targets both VEGFA and PIK3R2. Mol Cell Biochem 2011, 351:157-64.
• [17]Fish JE, Santoro MM, Morton SU, Yu S, Yeh RF, Wythe JD, Ivey KN, Bruneau BG, Stainier DY, Srivastava D: miR-126 regulates angiogenic signaling and vascular integrity. Dev Cell 2008, 15:272-84.
• [18]Wang S, Aurora AB, Johnson BA, Qi X, McAnally J, Hill JA, Richardson JA, Bassel-Duby R, Olson EN: The endothelial-specific microRNA miR-126 governs vascular integrity and angiogenesis. Dev Cell 2008, 15:261-71.
• [19]Guo C, Sah JF, Beard L, Willson JK, Markowitz SD, Guda K: The noncoding RNA, miR-126, suppresses the growth of neoplastic cells by targeting phosphatidylinositol 3-kinase signaling and is frequently lost in colon cancers. Genes Chromosomes Cancer 2008, 47:939-46.
• [20]Sessa R, Seano G, di Blasio L, Gagliardi PA, Isella C, Medico E, Cotelli F, Bussolino F, Primo L: The miR-126 regulates angiopoietin-1 signaling and vessel maturation by targeting p85beta. Biochim Biophys Acta 1823, 2012:1925-35.
• [21]Harris TA, Yamakuchi M, Ferlito M, Mendell JT, Lowenstein CJ: MicroRNA-126 regulates endothelial expression of vascular cell adhesion molecule 1. Proc Natl Acad Sci U S A 2008, 105:1516-21.
• [22]Sun Y, Bai Y, Zhang F, Wang Y, Guo Y, Guo L: miR-126 inhibits non-small cell lung cancer cells proliferation by targeting EGFL7. Biochem Biophys Res Commun 2010, 391:1483-9.
• [23]Li XM, Wang AM, Zhang J, Yi H: Down-regulation of miR-126 expression in colorectal cancer and its clinical significance. Med Oncol 2011, 28:1054-7.
• [24]Li X, Zhang G, Luo F, Ruan J, Huang D, Feng D, Xiao D, Zeng Z, Chen X, Wu W: Identification of aberrantly expressed miRNAs in rectal cancer. Oncol Rep 2012, 28:77-84.
• [25]Hansen TF, Soerensen FB, Lindebjerg J, Jakobsen A: The predictive value of microRNA-126 in relation to first line treatment with capecitabine and oxaliplatin in patients with metastatic colorectal cancer. BMC Cancer 2012, 12:83. doi:10.1186/1471-2407-12-83 BioMed Central Full Text
• [26]Schepeler T, Reinert JT, Ostenfeld MS, Christensen LL, Silahtaroglu AN, Dyrskjot L, Wiuf C, Sorensen FJ, Kruhoffer M, Laurberg S, Kauppinen S, Orntoft TF, Andersen CL: Diagnostic and prognostic microRNAs in stage II colon cancer. Cancer Res 2008, 68:6416-24.
• [27]Hansen TF, Nielsen BS, Jakobsen A, Sorensen FB: Visualising and quantifying angiogenesis in metastatic colorectal cancer: A comparison of methods and their predictive value for chemotherapy response. Cell Oncol 2013, 36:341-50.
• [28]Moore HM, Kelly AB, Jewell SD, McShane LM, Clark DP, Greenspan R, Hayes DF, Hainaut P, Kim P, Mansfield EA, Potapova O, Riegman P, Rubinstein Y, Seijo E, Somiari S, Watson P, Weier HU, Zhu C, Vaught J: Biospecimen reporting for improved study quality (BRISQ). Cancer Cytopathol 2011, 119:92-101.
• [29]Jorgensen S, Baker A, Moller S, Nielsen BS: Robust one-day in situ hybridization protocol for detection of microRNAs in paraffin samples using LNA probes. Methods 2010, 52:375-81.
• [30]Nielsen BS, Jorgensen S, Fog JU, Sokilde R, Christensen IJ, Hansen U, Brunner N, Baker A, Moller S, Nielsen HJ: High levels of microRNA-21 in the stroma of colorectal cancers predict short disease-free survival in stage II colon cancer patients. Clin Exp Metastasis 2011, 28:27-38.
• [31]Schmidt MH, Bicker F, Nikolic I, Meister J, Babuke T, Picuric S, Muller-Esterl W, Plate KH, Dikic I: Epidermal growth factor-like domain 7 (EGFL7) modulates Notch signalling and affects neural stem cell renewal. Nat Cell Biol 2009, 11:873-80.
• [32]Pinte SB, Soncin F: Egfl7 promotes tumor escape from immunity. Oncoimmunology 2012, 1:375-6.
• [33]Nikolic I, Stankovic ND, Bicker F, Meister J, Braun H, Awwad K, Baumgart J, Simon K, Thal SC, Patra C, Harter PN, Plate KH, Engel FB, Dimmeler S, Eble JA, Mittelbronn M, Schafer MK, Jungblut B, Chavakis E, Fleming I, Schmidt MH: EGFL7 ligates alphavbeta3 integrin to enhance vessel formation. Blood 2013, 121:3041-50.
• [34]Hansen TF, Andersen CL, Nielsen BS, Spindler KL, Sorensen FB, Lindebjerg J, Brandslund I, Jakobsen A: Elevated microRNA-126 is associated with high vascular endothelial growth factor receptor 2 expression levels and high microvessel density in colorectal cancer. Oncology letters 2011, 2:1101-6.
• [35]Saito Y, Friedman JM, Chihara Y, Egger G, Chuang JC, Liang G: Epigenetic therapy upregulates the tumor suppressor microRNA-126 and its host gene EGFL7 in human cancer cells. Biochem Biophys Res Commun 2009, 379:726-31.
• [36]Zhang Y, Yang P, Sun T, Li D, Xu X, Rui Y, Li C, Chong M, Ibrahim T, Mercatali L, Amadori D, Lu X, Xie D, Li QJ, Wang XF: miR-126 and miR-126* repress recruitment of mesenchymal stem cells and inflammatory monocytes to inhibit breast cancer metastasis. Nat Cell Biol 2013, 15:284-94.
• [37]Sasahira T, Kurihara M, Bhawal UK, Ueda N, Shimomoto T, Yamamoto K, Kirita T, Kuniyasu H: Downregulation of miR-126 induces angiogenesis and lymphangiogenesis by activation of VEGF-A in oral cancer. Br J Cancer 2012, 107:700-6.
• [38]Png KJ, Halberg N, Yoshida M, Tavazoie SF: A microRNA regulon that mediates endothelial recruitment and metastasis by cancer cells. Nature 2011, 481:190-4.
• [39]Tavazoie SF, Alarcon C, Oskarsson T, Padua D, Wang Q, Bos PD, Gerald WL, Massague J: Endogenous human microRNAs that suppress breast cancer metastasis. Nature 2008, 451:147-52.