|Year : 2019 | Volume
| Issue : 1 | Page : 159-164
Significance of epithelial cell adhesion molecule and epidermal growth factor receptor expression in papillary thyroid carcinoma: an immunohistochemical study
Rasha M Abdrabh, Heba M Rashad MD
Pathology Department, Faculty of Medicine, Benha University, Banha, Egypt
|Date of Submission||30-Jul-2019|
|Date of Acceptance||20-Aug-2019|
|Date of Web Publication||29-Nov-2019|
Heba M Rashad
Pathology Department, Faculty of Medicine, Benha University, Banha, 11311
Source of Support: None, Conflict of Interest: None
Background Thyroid carcinoma ranks as the most common among malignancies of endocrine organs, and a rapid increase in its incidence has been observed over the years. Assessment of epithelial cell adhesion molecule (EpCAM) and epidermal growth factor receptor (EGFR) immunopositivity in papillary carcinoma thyroid was conducted, and expression was correlated with the clinicopathological parameters.
Materials and methods Immunohistochemical detection of EpCAM and EGFR was investigated in 33 thyroidectomy specimens with papillary carcinoma and 10 cases of hyperplastic thyroid nodule. The association between those markers was studied, besides their correlation with clinicopathological parameters.
Results Both EpCAM and EGFR showed weak or low expression in hyperplastic nodule compared with higher expression in papillary carcinoma (P<0.001). In cases showing extrathyroid extension, lymph node metastasis, distant metastasis, and advanced TNM stage, EpCAM and EGFR were significantly correlated (P<0.001, 0.01, 0.01, <0.001, 0.02, <0.001, 0.02, 0.02, and <0.001, respectively).
Conclusion Combined expression of EpCAM and EGFR may be related to papillary thyroid carcinogenesis, which could be used as therapy for cancer management.
Keywords: epidermal growth factor receptor, epithelial cell adhesion molecule, papillary thyroid carcinoma
|How to cite this article:|
Abdrabh RM, Rashad HM. Significance of epithelial cell adhesion molecule and epidermal growth factor receptor expression in papillary thyroid carcinoma: an immunohistochemical study. Egypt J Pathol 2019;39:159-64
|How to cite this URL:|
Abdrabh RM, Rashad HM. Significance of epithelial cell adhesion molecule and epidermal growth factor receptor expression in papillary thyroid carcinoma: an immunohistochemical study. Egypt J Pathol [serial online] 2019 [cited 2020 Jul 6];39:159-64. Available from: http://www.xep.eg.net/text.asp?2019/39/1/159/271998
| Introduction|| |
Regarding the incidence of thyroid carcinoma, a rapid rise has been observed particularly in women, reaching to 16.3/100 000 in 2008 (Davies and Welch, 2014).
In the USA, thyroid carcinoma represents nearly 1% of all cancers and ∼0.2% of cancer deaths (Torre et al., 2016).
In Egypt, at NCI pathology cancer registry, primary malignant thyroid neoplasm constituted 1.97% of all malignant neoplasm, 74.7% of malignant neoplasm of endocrine system, and 38.67% of all malignant lesions (El Bolkiny et al., 2016).
Among different types of thyroid carcinoma, papillary thyroid carcinoma accounted for ∼90% of thyroid carcinoma cases (Sofiadis et al., 2009).
For thyroid carcinoma to develop, there are few well-known established risk factors, including history of ionizing radiation exposure, and some rare familial syndromes (Ron and Schneider, 2006).
The epithelial cell adhesion protein (EpCAM) belongs to a family that is responsible for cell adhesion (Ralhan et al., 2010). Physiologically, in epithelial cells except squamous epithelia, the expression pattern of EpCAM is mainly expressed in the intercellular space, whereas in connective tissue, it is not expressed. Overexpression of EpCAM was strongly correlated with decreased overall survival and poor prognostic outcome in various types of tumors (Terris et al., 2010).
Epidermal growth factor (EGFR) belongs to human epidermal receptor family (Uberall et al., 2008). Once activated, homodimeric or heterodimeric changes occur, which eventually activate the intrinsic tyrosine kinase, triggering multiple signaling pathways (Hudson et al., 2009). These pathways influence many behaviors of cancer cells, with disturbance of cell proliferation and cohesion, and also enable cells to escape apoptosis and accelerate the migration and invasion of tumor cells (Yarden and Pines, 2012).
This research was performed to assess the immunohistochemical expression of EpCAM and EGFR in papillary thyroid carcinoma and the possible correlation with the clinicopathological parameters.
| Materials and methods|| |
This performed retrospective study included 33 thyroidectomy specimens with papillary carcinoma and 10 cases of hyperplastic thyroid nodule. They were gathered from Archives of Pathology Department and Early Cancer Detection Unit, Faculty of Medicine, Benha University, during the years 2009–2018. The clinicopathologic characteristics, regarding age, sex, lymph node status, extracapsular extension, and distant metastasis status were taken for every case. The study was approved by the Ethical Committee in Faculty of Medicine, Benha University.
Reexamination of previously stained slides with hematoxylin and eosin was done to confirm the previous diagnosis and to assess the pathological T stage and lymph node metastasis. TNM stage was defined according to American Joint Committee on Cancer on Cancer Criteria Tuttle et al., (2017).
For immunohistochemical processing, sections of 4-μm thickness were cut and prepared on charged slides. Xylene was used to deparaffinize sections, and then treated with descending grades of ethanol, followed by treatment with 0.3% H2O2 for 5 min. A citrate buffer (pH 6) was used for the process of antigen retrieval. Incubation with EGFR (rabbit polyclonal, dilution 1 : 200; Novus Bio, Novus Biologicals, LLC 8100 Southpark Way, A-8 Littleton CO, USA) and EpCAM [mouse, monoclonal antibody, ready to use; Thermoscientific, Waltham, Massachusetts, USA; cat. #MS-144-R7 (0.7 ml)] was performed overnight at 4°C. Diaminobenzidine and hematoxylin were used as a chromogen and counter stain, respectively.
Positive controls for EGFR and EpCAM were cutaneous squamous cell carcinoma and breast carcinoma, respectively. Omitting the primary antibody and replacing it with normal rabbit serum IgG was used as a negative control for each marker.
Regarding EpCAM, membranous and cytoplasmic expression is considered positive. The intensity of the stained cells ranged from 0 to 3+. A scoring system of positively stained tumor cells was used and ranged from 0 to 100%. Multiplication of the two previous scores was done to yield a final expression score: less than 3 is considered a negative score, from 3 to 5 is considered weak positive, from 6 to 8 is considered moderately positive, and more than 9 is strongly positive (Yanamoto et al., 2007).
Regarding EGFR, staining intensity ranged from 0 to 3+ and the percent of positivity ranged from 0 to 100%. Low EGFR is applied for 1+ intensity regardless of the percent of positivity or less than 50% of tumor cells showed positivity with 2+ intensity. High EGFR is applied for any percent of positivity with 3+ intensity (Fisher et al., 2013a, Fisher et al., 2013b).
All analyzed data were carried out using STATA/SE version 11.2 for Windows (STATA Corporation, College Station, Texas, USA).
P value less than 0.05 was considered as statistically significant, less than 0.001 is highly significant, whereas more than 0.05 is nonsignificant.
| Results|| |
Clinicopathological data of patients
The studied 33 cases of papillary thyroid carcinoma included five (15%) males and 28 (85%) females. All clinicopathological data are summarized in [Table 1].
Expression of epithelial cell adhesion molecule and epidermal growth factor receptor in the studied cases
Weak or low expression for both EpCAM and EGFR was noted in hyperplastic nodule compared with higher expression in carcinoma (P<0.001; [Table 2] and [Figure 1],[Figure 2],[Figure 3]).
|Table 2 Epithelial cell adhesion molecule and epidermal growth factor receptor expression in the studied cases|
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|Figure 1 Hyperplastic nodule with negative expression of epithelial cell adhesion molecule (IHC, ×200).|
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|Figure 2 Papillary thyroid carcinoma with strong membranous expression of epithelial cell adhesion molecule (IHC, ×400).|
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|Figure 3 Papillary thyroid carcinoma with high epidermal growth factor receptor membranous expression (IHC, ×200).|
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Correlation of epithelial cell adhesion molecule with the clinicopathological parameters
In cases showing extrathyroid extension, lymph node metastasis, distant metastasis, and advanced TNM stage, EpCAM was significantly correlated (P<0.001, 0.01, 0.01 and <0.001, respectively). Regarding other variables, no significant correlation was illustrated ([Table 3]).
|Table 3 Correlation of epithelial cell adhesion molecule with clinicopathological parameters|
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Correlation of epidermal growth factor receptor to clinicopathological parameters
High EGFR expression was detected in cases with capsular invasion, extrathyroid extension, lymph node metastasis, distant metastasis, and advanced TNM stage, and this was of statistical significance (P=0.02, <0.001, 0.02, 0.02 and <0.001, respectively). Concerning correlation of EGFR with other variables such as age, sex, tumor size, tumor site, and lymphatic/vascular invasion, no significant correlation was detected ([Table 4]).
|Table 4 Correlation of epidermal growth factor receptor with clinicopathological parameters|
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Correlation of epithelial cell adhesion molecule and epidermal growth factor receptor expression in studied cases
To detect if EpCAM expression is positively correlated with EGFR in the examined cases, Fisher exact test was used. Of 33 cases of papillary thyroid carcinoma, 15 cases showed higher expression for both EpCAM and EGFR, denoting a significant positive correlation (P<0.001; [Table 5]).
|Table 5 Correlation between epithelial cell adhesion molecule and epidermal growth factor receptor expression in studied cases:|
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| Discussion|| |
Previous studies demonstrated that many cell adhesion molecules are responsible for tumor genesis and metastasis (Kroepil et al., 2013).
In this study, EpCAM expression was noted in both carcinoma cases and hyperplastic thyroid nodule localized at the cell surface membrane. Its expression was weak in the hyperplastic nodule compared with strong expression in papillary thyroid carcinoma (P<0.001). These results were compatible with Ralhan et al. (2010) and Okada et al. (2014) during their immunohistochemical analysis of thyroid carcinoma. They revealed that in the well-differentiated cancers, EpCAM was strongly expressed. Moreover, this finding is in concordance with the previous study done on colon carcinomas series that revealed higher expression of EpCAM in tumor tissues in contrast to poor expression in tissues adjacent to carcinoma and noncancerous tissues (Went et al., 2006).
This variation in expression scoring of EpCAM between normal tissue and carcinoma ones can be explained by that EpCAM directly affects the cell cycle control and accelerates growth of cells through the upregulation of many genes including the genes that control cell cycle, the c-myc proto-oncogene, and the activation of the Wnt signaling pathway, which eventually lead to tumor formation (Wang et al., 2013).
Regarding the correlation with clinicopathological parameters, a significantly positive correlation between EpCAM expression and extrathyroid extension (P<0.001), lymph node status (P=0.01), distant metastasis (P=0.01), and TNM stage (P<0.001) was illustrated. These findings were in agreement with Du et al. (2010), Ni et al. (2012) and Chan et al. (2014) in their series of gastric carcinoma, prostatic carcinoma, and hepatocellular carcinoma, respectively. These results can be interpreted by the ability of EpCAM to disturb the link between α-catenin and cytoskeleton with loss of E-cadherin that mediate cell adhesion, resulting in facilitation of cell motility and invasion (Cheng et al., 2015).In the studied carcinoma cases, a significantly high score of EGFR was detected than in nodular hyperplasias. That pattern of EGFR expression was in line with Dai et al. (2017) and Gong et al. (2012), suggesting that the increased expression EGFR is associated with the occurrence of carcinoma. The overexpression of EGFR disturbs regulated cell cycle through continuous transmission of signals to cells, which lead to continuous proliferation and malignant transformation.
This work demonstrated a higher EGFR expression in cases with capsular invasion, extrathyroid extension, lymph node status, distant metastasis, and TNM stage, and this was of statistical significance (P=0.02, <0.001, 0.02, 0.02, and <0.001, respectively). These results indicate the association of EGFR with aggressive behaviors in thyroid carcinoma. This was compatible with Wu et al. (2003), Fisher et al., (2013a), Fisher et al., (2013b), and Dai et al. (2017). Our results further suggest that cases of papillary thyroid carcinoma with high EGFR expression may have a role in metastasis and invasion of tumor cells.
In this work, we illustrated that EpCAM was positively correlated with EGFR expression in papillary thyroid carcinoma (P<0.001), suggesting the possible combined role of EpCAM and EGFR for development of carcinoma. These results were in concordance with that obtained by Schartinger et al. (2009) in squamous cell carcinoma and Zheng et al. (2017) in epithelial ovarian carcinoma.
| Conclusion|| |
EpCAM and EGFR overexpression is associated with a possible malignant potential. This combined expression could represent promising biomarkers for the development and progression of papillary thyroid carcinoma and may be used as potential molecular therapeutic targets.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
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[Figure 1], [Figure 2], [Figure 3]
[Table 1], [Table 2], [Table 3], [Table 4], [Table 5]