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High Levels of EGFR Expression in Tumor Stroma Are Associated with Aggressive Clinical Features in Epithelial Ovarian Cancer

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High Levels of EGFR Expression in Tumor Stroma Are Associated with Aggressive Clinical Features in Epithelial Ovarian Cancer Journal name: OncoTargets and Therapy Article Designation: Original Research Year: 2016 Volume: 9 OncoTargets and Therapy Dovepress Running head verso: Wang et al Running head recto: Clinical significance and biological function of EGFR open access to scientific and medical research DOI: http://dx.doi.org/10.2147/OTT.S96309 Open Access Full Text Article ORIGINAL RESEARCH High levels of EGFR expression in tumor stroma are associated with aggressive clinical features in epithelial ovarian cancer Ke Wang Purpose: The aim of this study was to investigate the clinical significance and biological Dan Li function of epidermal growth factor receptor (EGFR) expressed in tumor stroma of epithelial Lu Sun ovarian cancer. Methods: Immunohistological staining of EGFR was evaluated in 242 patients with epithelial Department of Gynecologic Cancer, National Clinical Research Center ovarian cancer. The correlations of EGFR expression in tumor stroma with clinicopathological for Cancer, Tianjin Key Laboratory features and with the expression level of Ki-67 were analyzed by SPSS software. Kaplan–Meier of Cancer Prevention and Therapy, analysis and the Cox proportional hazard model were used to analyze the effect of EGFR Tianjin Medical University Cancer Institute and Hospital, Tianjin, People’s expression in tumor stroma on the prognosis of patients with epithelial ovarian cancer. Mean- Republic of China while, the activities of proliferation and migration of tumor cells were detected when EGFR For personal use only. overexpressed in stroma cells. Results: EGFR expression in tumor stroma correlated significantly with clinical stage (χ2=7.002, P=0.008) and distant metastases (χ2=16.59, P,0.001). Furthermore, there was a significantly positive correlation between the level of EGFR expressed in tumor stroma and the level of Ki-67 expressed in tumor cells (χ2=6.120, P=0.013). Patients with high EGFR expression level in tumor stroma showed poor survival (P=0.002). Multivariate analysis showed that high expression of EGFR in tumor stroma was an independent predictor for epithelial ovarian cancer patients (hazard ratio =1.703; 95% confidence interval 1.125–2.578, P=0.012). Furthermore, stroma cells overexpressing EGFR could promote the proliferation and migration of adjacent tumor cells. Conclusion: High expression of EGFR in tumor stroma correlates with aggressive clinical OncoTargets and Therapy downloaded from https://www.dovepress.com/ by 54.70.40.11 on 11-Dec-2018 features in epithelial ovarian cancer, and is an independent prognostic factor. Keywords: EGFR, epithelial ovarian cancer, tumor stroma, clinical features, overall survival, prognostic factor Introduction Since ovarian cancer, located deep within the pelvis, has no early typical symptoms, it is difficult to detect at an early stage. Because of the lack of effective therapies for advanced-stage disease, epithelial ovarian cancer is the deadliest gynecological malig- nancy and the second leading cause of cancer-related deaths among women worldwide.1 Correspondence: Ke Wang About 22,240 women were diagnosed with invasive epithelial ovarian cancer in the Department of Gynecologic Cancer, 2 National Clinical Research Center for USA in 2013. In ovarian cancer, disease histotype, differentiation grade, age, and Cancer, Tianjin Key Laboratory of Cancer performance status are well-known clinicopathological prognostic factors.3 Although Prevention and Therapy, Tianjin Medical these parameters can reflect biological features of patients, they are not sensitive or University Cancer Institute and Hospital, Huanhuxi Road, Hexi District, Tianjin sufficiently specific for the individual. Therefore, it is urgent to find new biomarkers, 300060, People’s Republic of China which should aid in a more accurate prediction of survival and therapeutic targets for Tel +86 138 2108 1782 Email [email protected] patients with epithelial ovarian cancer.4,5 submit your manuscript | www.dovepress.com OncoTargets and Therapy 2016:9 377–386 377 Dovepress © 2016 Wang et al. This work is published and licensed by Dove Medical Press Limited. The full terms of this license are available at https://www.dovepress.com/terms.php and incorporate the Creative Commons Attribution – Non Commercial (unported, v3.0) License (http://creativecommons.org/licenses/by-nc/3.0/). By accessing the work you http://dx.doi.org/10.2147/OTT.S96309 hereby accept the Terms. Non-commercial uses of the work are permitted without any further permission from Dove Medical Press Limited, provided the work is properly attributed. For permission for commercial use of this work, please see paragraphs 4.2 and 5 of our Terms (https://www.dovepress.com/terms.php). Powered by TCPDF (www.tcpdf.org) 1 / 1 Wang et al Dovepress Ovarian surface epithelium (OSE) is a single layer of epi- Table 1 Correlations between EGFR expression in tumor cells thelial cells in the surface of the ovary.6 The stroma of ovarian and clinicopathological characters tissue can produce growth factors and cytokines, which act Variables n n χ2 P-value on the OSE and maintain the normal function of the ovary.7 EGFR-tc EGFR-tc The altered cellular activity of the OSE contributes to the low high etiology of ovarian cancer, and the stroma play an important Age (years) ,55 122 56 66 1.122 0.289 8 role in this process. Tumor invasion also requires an asso- $55 120 47 73 ciation with stromal tissue and most ovarian tumors have a Clinical stage stromal-like component.9 Therefore, stromal–epithelial cell Early (stage I–II) 93 50 43 7.752 0.005 interactions appear to have a critical role in the function and Advanced (stage III–IV) 149 53 96 Histological grade growth of ovarian cancer. The tumor stroma is increasingly I 38 17 21 0.728 0.695 perceived as a major contributor to the pathogenesis and II 58 27 31 disease progression in practically all cancer types.10 III 146 59 87 Epidermal growth factor receptor (EGFR) is one of the Ascites No 90 38 52 0.007 0.934 receptor tyrosine kinases, mediating responses of extracellular Yes 152 65 87 signals to control cell differentiation, proliferation, and migra- Metastases tion, expressed in both tumor cells and tumor stroma.11 EGFR Negative 64 40 24 14.149 ,0.001 Positive 178 63 115 holds considerable promise as a therapeutic target.12 Not Notes: EGFR expression in tumor stroma and its correlation with clinicopathological surprisingly, there are also many published papers attempting variables. Bold values indicate P,0.05. assess the relationship between EGFR overexpression and Abbreviations: EGFR, epidermal growth factor receptor; EGFR-tc, EGFR expressed in tumor cells. survival. However, the data regarding the prognostic role of EGFR expression are inconsistent.13 Many researchers are specifically concerned with EGFR expressed in tumor cells, Inc., Dallas, TX, USA), anti-Ki-67 (ab16667; Abcam, For personal use only. but EGFR expressed in tumor stroma attracts little attention. Cambridge, UK), anti-E-cadherin (ab1416; Abcam), antivi- We report here that high expression of EGFR in tumor stroma mentin (ab92547; Abcam), anti-β-actin (sc-1616; Santa Cruz is associated with aggressive clinical features, and is a new Biotechnology), and antigreen fluorescence protein (GFP; prognosis marker for epithelial ovarian cancer patients. sc-8334; Santa Cruz Biotechnology). Antimouse antibodies (Santa Cruz Biotechnology) and antirabbit antibodies Materials and methods (Zhongshan Goldbridge Biotechnology, Beijing, China) were Patients and tissue samples used for Western blot in this study. Two hundred forty-two epithelial ovarian cancer tissue sections were obtained from the Department of Pathol- Immunohistochemistry staining ogy, Tianjin Cancer Hospital, Tianjin Medical University dur- OncoTargets and Therapy downloaded from https://www.dovepress.com/ by 54.70.40.11 on 11-Dec-2018 Tissue sections were first deparaffinized and rehydrated ing 2005–2007, and all the patients received surgical therapy with xylene and graded alcohol solutions, and then endog- and similar chemotherapy (Taxol/cisplatin or paclitaxel/cispla- enous peroxidase activity was quenched by 3% hydrogen tin). Written informed consent was obtained from all patients, peroxide, followed by boiling in 10 mM citrate buffer and the study was approved by the Ethical Committee of (pH 6.0) for 3 minutes in an autoclave sterilizer. After Tianjin Cancer Hospital. All tissue sections were examined by rinsing with phosphate-buffered saline (PBS), sections specialists to make a final diagnosis. The classification of can- were incubated with primary antibodies diluted 1:100 in cer stage and grade was according to the International Federa- antibody diluent (Zhongshan Goldbridge Biotechnology) tion of Gynecology and Obstetrics (2009). Clinicopathological overnight at 4°C. After rinsing with PBS, sections were data were collected including age, histology type, pathological incubated with secondary antibody PV6001 or PV6002 grade, ascites, metastasis status, and tumor clinical stage. All (Zhongshan Goldbridge Biotechnology) for 1 hour at 37°C patients’ characteristics are summarized in Table 1. and stained with 3,3′-diaminobenzidine. The slides were counterstained with hematoxylin, dehydrated with ethanol, Antibodies cleared with xylene, and mounted in neutral gum. Nega- The primary antibodies used in this study are listed as tive control sections were incubated with PBS instead of follows: anti-EGFR (sc-03; Santa Cruz Biotechnology a primary antibody. 378 submit your manuscript | www.dovepress.com OncoTargets and Therapy
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