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Abstract

Pleomorphic adenoma (PA) is the most common salivary gland neoplasm and, although mostly benign, recurrences, being called recurrent pleomorphic adenoma (RPA) and malignant transformation to carcinoma ex pleomorphic adenoma (CXPA), do occur. Recently, attention has been focused on molecular targeted cancer therapy in various tumors, including salivary gland tumors. The aim of this study was to investigate the role of estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor-2 (HER-2) in PA, RPA, and CXPA. In total, 20 cases of PA, 18 of RPA, and 7 cases of CXPA were immunohistochemically studied for ER, PR, and HER-2. For evaluation of ER and PR, only nuclear expression and greater than 10% positive cells were regarded as cutoff criteria. HER-2 was evaluated semiquantitatively and graded from 0 to 3+. HER-2 amplification was assessed by chromogenic in situ hybridization (CISH). Tumors were negative for ER, PR, and HER-2 in all cases of PA and RPA. A case of CXPA showed moderate and complete membranous staining, and 6 cases were negative. HER-2 amplification was not observed in any case. In conclusion, the lack of ER, PR, and HER-2 expression in PA, RPA, and CXPA suggests that these proteins are not involved in progression, recurrence, or malignant transformation of PA.

Keywords

Pleomorphic adenoma recurrence estrogen progesterone HER-2

Introduction

Pleomorphic adenoma (PA) is the most common neoplasm of salivary glands.¹ PA is classified as a benign tumor; however, the incidence of recurrence (RPA) after initial operative treatment is significant and varies largely because of differences in surgical technique.^2,3 Recurrences have been associated with an increased risk of malignant transformation to carcinoma ex pleomorphic adenoma (CXPA).^4,5

Some factors are related to the increase in the recurrence, such as multinodularity and pseudopodia, the age of the patients, and incomplete surgical excision.^6,7 Recurrence has also been associated with cell biological and molecular changes.^7-12

In previous studies from our group, several aspects of RPA have been performed and important results were obtained. Hamada et al⁷ and Soares et al¹⁰ observed that Mucin 1 (MUC 1), a glycoprotein that plays a role in homeostasis and carcinogenesis, is related to recurrence and malignant transformation of PA. Similar results were observed with fibroblast growth factor and platelet-derived growth factor (PDGF), in which the RPA presented a higher immunohistochemical expression of these factors when compared with PA.⁸ The p16, cyclin D1, and E2F proteins, which compound the retinoblastoma pathway that controls cell cycle phases, also showed strong expression in RPA. These results show that cell cycle-related changes in RPA are similar to changes in CXPA.¹²

Estrogen and progesterone are steroid hormones responsible for many biological processes with the potential to cause specific changes in anatomy and physiology throughout human development.¹³ Estrogen and progesterone function through the interaction between hormones and their respective receptor proteins, namely estrogen (ER) and progesterone (PR) receptors, respectively. Regarding ER, there are 2 described receptors, named ERα and ERβ, where the ERα is the most common isoform studied.¹⁴ ER and PR are commonly implicated in cell growth by hormone-induced cell proliferation.¹⁵ Human epidermal growth factor receptor-2 (HER-2) is a proto-oncogene present on chromosome 17q and it is overexpressed in a variety of malignancies.¹⁶ HER-2 is associated with increased vessel permeability, endothelial cell growth, proliferation, migration, and differentiation.^17,18

The participation of ER, PR, and HER-2 in the pathogenesis and development of tumors is widely documented, especially in breast, endometrium, and prostate tumors.^19-21 In breast tumors, ER, PR, and HER-2 expression assists in the choice of treatment. In general, hormone-dependent lesions have better prognosis compared with nonhormone-dependent tumors.²² The involvement of ER, PR, and HER-2 is also described in benign and malignant salivary gland tumors.^15,23-34 A few studies on hormone and HER-2 therapy to treat malignant tumors of the salivary gland have been published.^35-38

The management of RPA is challenging due to the multiple nodules that may add up to as many as 130 in a single patient,⁹ which hinders complete excision of the lesion thus favoring future recurrences and malignant transformation. In such scenarios, one wonders whether hormone therapy could be a useful approach in the management of RPA. Therefore, the aim of this study was to investigate the role of ER, PR, and HER-2 in PA, RPA, and CXPA.

Materials and Methods

This study examined 20 cases of PA without recurrence, 18 cases of RPA, and 7 cases of CXPA. The PA group included 12 women and 8 men. The tumors were located in the parotid gland in 17 cases and in the submandibular gland in 3 cases. The mean age was 43 years. The RPA group included 8 women and 10 men, the mean age was 42 years, and the tumors were located in the parotid glands in 17 cases and in the submandibular gland in 1 case. Information was not available for 1 case of RPA. The CXPA group included 4 men and 2 women, and the mean age was 64 years. The tumors were situated in the parotid gland in 4 cases and in the minor salivary glands in 2 cases. Information was not available for 1 case of CXPA.

Four cases of CXPA were recurrences of previous PA, and in 3 cases, this information was not available. The diagnosis of CXPA was based on histopathologic characteristics and classified as noninvasive and invasive. All studied cases showed malignant transformation of the PA luminal cells. Five cases of CXPA were noninvasive and 2 cases were invasive.

The median time interval between initial operation and recurrence (RPA) was 10.6 years (range of 1-25 years), with 2 (11.11%) cases with a first recurrence within 5 years, 2 (11.11%) after 5 years, and 9 (50%) after more than 10 years. Information was not available for 5 (27.78%) patients.

Immunohistochemistry

Three-μm sections were taken from each block and mounted on aminopropyltriethoxysilane-coated slides. Following deparaffinization in xylene and rehydration through decreasing concentrations of ethanol, endogenous peroxidases were inhibited using 3% H₂O₂ in methanol at room temperature. Immunohistochemistry for ERα and PR was subsequently performed using the antibodies and detection methods as specified in Table 1. Immunoreactivity was observed with 3.3′-diaminobenzidine tetrahydrochloride (DAB, 5 minutes at 37°C), and the slides are counterstained with Mayer hematoxylin. Immunohistochemistry staining for HER-2 protein was performed using the HercepTest Kits as recommended by the manufacturer (DakoCytomation).

Table 1.

Details of the antibodies used for immunohistochemistry.

Antibodies	Clone	Dilution	Source	Buffer (AR)
ER	1D5	1:150	Dako^a	EDTA
PR	PgR 636	1:300	Dako^a	EDTA
HercepTest			Dako^a

Abbreviations: ER, estrogen receptor; PR, progesterone receptor; EDTA: ethylenediaminetetraacetic acid.

Dako Corporation, Glostrup, Denmark.

Evaluation of immunohistochemistry staining

ER, PR, and HER-2 immunohistochemical reactions were interpreted by 2 authors. Only nuclear immunoreactivity was considered positive to ER and PR. Cases were considered positive for ER and PR according to standardized guidelines using a cutoff of >10% stained tumor nuclei.^39,40 Only membranous staining was considered positive to HER-2, and the staining was scored semiquantitatively using the HercepTest protocol (Dako, Carpinteria, CA, USA) where the grades are 0 to 3+. Score 0 = no staining or staining in <10% of tumor cells; score 1, faint and partial staining in ⩾10% of tumor cells; score 2, weak-to-moderate complete membrane staining in ⩾10% of tumor cells; and score 3, strong complete membrane staining in ⩾10% of tumor cells.⁴¹ Breast carcinomas cases were used as positive control in all reactions.

CISH assessment of HER-2 status

Chromogenic in situ hybridization (CISH) to HER-2 was realized according to the manufacturer’s protocol (Dako Her2 CISH pharmDx kit). Areas containing the highest HER-2 counts were identified by counting HER-2 and centromeric probe 17 (CEP17) in at least 20 nuclei. Only nonoverlapping nuclei with distinct nuclear borders were considered. The ratio between HER-2 and CEP17 was calculated, and the HER-2 gene was considered amplified when the ratio of gene-specific HER-2 to CEP17 signals was 2.0 or more or when an HER-2 signal cluster was observed. At least 1 CISH-positive spot was needed to assign a case to the HER-2-amplified category. When assessing intratumor variability, the results of CISH were considered separately for each core.

Statistical analysis

For comparison between the different tumor types, the Mann-Whitney test was used. Results with P < 0.05 were considered significant.

Results

Estrogen receptor

In all tumor groups, ER immunoreactivity was observed only in a few myoepithelial and ductal cells, though most cells were negative. In PA, ER was negative in 19 cases, whereas 1 case showed more than 10% positive cells. For RPA, 18 cases were negative (Figures 1 and 2). The difference between PA and RPA was not statistically significant (Mann-Whitney test; P = 1.000).

Figure 1.

ER negative expression in PA (A), RPA (B), and CXPA (C). PR negative expression in PA (D), RPA (E), and CXPA (F). HER-2 negative expression in PA (G), RPA (H), and CXPA (I). PA indicates pleomorphic adenoma; RPA, recurrent pleomorphic adenoma; CXPA, carcinoma ex pleomorphic adenoma; ER, estrogen receptor; PR, progesterone receptor.

Figure 2.

Graphics illustrating the expression of ER, PR, and HER-2 in PA, RPA, and CXPA, respectively. ER indicates estrogen receptor; PR, progesterone receptor; PA, pleomorphic adenoma; RPA, recurrent pleomorphic adenoma; CXPA, carcinoma ex pleomorphic adenoma.

All cases of CXPA were also negative for ER (Figure 1), though in 1 case of the remaining RPA, greater than 10% positivity was observed focally. In the normal glandular tissue, no staining was observed in ductal and acinar cells.

Progesterone receptor

All cases of PA and RPA showed less than 10% positivity for PR (Figure 1). The Mann-Whitney test revealed no significant difference for PR expression (P = 1.000) between PA and RPA (Figure 2). All CXPA cases were also negative for PR (Figure 1), though greater than 10% positivity was observed focally in 1 case of the remaining RPA. In the normal glandular tissue, a subtle staining was observed in acinar cells.

Human epidermal growth factor receptor-2

In tumors from groups PA and RPA, HER-2 protein immunoreactivity was negative (grade 0) (Figures 1 and 2). The Mann-Whitney test revealed no significant difference for HER-2 expression (P = 1.000) between PA and RPA. In CXPA, HER-2 was “grade 0” in 6 cases and 1 case was “grade 2” (Figures 1 and 2). The case with labeling grade 2 was classified as invasive CXPA.

Chromogenic in situ hybridization

HER-2 amplification was not seen in none of the PA, RPA, and CXPA cases.

Discussion

RPA is lesion with peculiar clinical and biological features. Although classified as a benign lesion, its clinical behavior could be aggressive, which increases the risk of recurrence and malignant transformation. In previous studies from our group, we have demonstrated that some biological features of RPA, such as tumor vascularization, cellular proliferation index, extracellular matrix, and some growth factors are similar to those of PA.^8,9 However, features such as overexpression of proteins of the retinoblastoma cell cycle pathway, MUC 1, PDGF-A, PDGF-B, PDGF-Rα, FGF-2, Bek, and Flg are similar to those observed in CXPA.⁷-12

ER and PR expression are well established in breast carcinoma, and their presence has been shown to confer a more favorable prognosis for the patients.⁴² Several studies have analyzed the participation of ER and PR in the pathogenesis of salivary gland tumors.²³-25,²⁸-33 In this study, PA cases presented predominantly negative markings for ER and PR. In the literature, in most of the studies, neither an ER nor a PR expression has been detected in a variety of benign tumors, such as PA, Warthin tumor, oncocytoma, basal cell adenoma, and myoepithelioma, demonstrating that these factors have no influence in the development tumors of the salivary glands.^{26,30,31,43,44} Although some authors have considered that the ERβ expression may have contributed to the development of PA, this antibody was not researched in this study.⁴⁵

In our study, ER and PR expression in all cases of RPA were negative, and the difference between the expression of ER in PA and RPA cases was not statistically significant. In the scientific literature, the knowledge about the role of ER and PR in RPA is scarce; only 1 study has been found. Glas et al¹⁵ found no significant difference in ER expression in cases of PA and RPA. The authors postulated that the expression of ER in both groups was not sufficient to expect any influence of ER on tumor growth. Regarding PR, Glas et al¹⁵ reported intense immunostaining in recurrent tumors. Of the cases examined only 2 were negative and about 27% of cases showed overexpression, with significant difference between cases of PA and RPA. The authors speculated that PR is a prognostic factor in the occurrence of recurrent pleomorphic adenoma. However, in our research, the difference between PR expression in PA and RPA was not significant, and we adopt that the hormone receptors not influence or predict recurrence of PA.

In all CXPA cases, the expression of ER and PR were negative, but curiously, 1 case showed moderate expression of both markers in the remaining PA area. Nasser et al²⁶ assessed ER and PR expression in cases of CXPA and not observed immunostaining in the tumors. The authors argue that larger studies take into account that the hormonal factors may be necessary for a more definitive assessment of hormonal receptors expression in salivary gland tumors.

HER-2 is a well-known epithelial tumor oncogene that encodes the epithelial growth factor receptor tyrosine kinase.³⁴ The expression of HER-2 has been identified in several types of human carcinoma, including breast, ovary, endometrial, and thyroid gland neoplasms, and has been associated with varied prognosis. In some salivary gland tumors, overexpression of the HER-2 protein has been shown with several outcomes.^{17,27,34,46-50}

We found immunostaining for HER-2 negative in PA, which corroborates some studies.^27,34,51-53 Our results and literature reports suggest that HER-2 expression changes are not related to the development of PA. Regarding RPA, there is only 1 study regarding HER-2 expression,⁵⁴ and no expression was observed. In this study, all cases of RPA were negative for HER-2. Based on our results and on other one found in the literature, we can postulate that there is no correlation between the expression of HER-2 and recurrence of tumors analyzed.

Overexpression and amplification of HER-2 was reported in some malignant salivary gland tumors as salivary duct carcinoma, adenocarcinoma NOS, mucoepidermoid carcinoma, and CXPA. Hashimoto et al³⁴ demonstrated that HER-2 superexpression appears to begin at the intraductal phase of CXPA and was mostly retained during the progression from intraductal to extracapsular components in individual extracapsular CXPA. Di Palma et al²⁷ also reported HER-2 expression in cases of intracapsular CXPA, and the authors hypothesize that HER-2 amplification and expression is an early event in malignant transformation of CXPA. Freitas et al⁴⁷ assessed HER-2 expression in CXPA cases, and HER-2 reactivity was observed almost exclusively in malignant luminal-type cells. The authors speculated that HER-2 could be a useful marker of malignant transformation in PA. In this study, 1 case of invasive CXPA, classified as epithelial histologically, presented labeling grade 2 for HER-2, showing that HER-2 expression could be related to malignant transformation.

In view of the results herein and those from previous studies regarding HER-2, RPA still has an expression profile closer to PA than to CXPA. In conclusion, the lack of ER, PR, and HER-2 expression in PA, RPA, and CXPA suggests that these proteins are not involved in progression, recurrence, or malignant transformation of PA.

Footnotes

Funding:

The author(s) disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This study was supported by the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) under number 2014/25173-8.

Declaration of Conflicting Interests:

The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Author Contributions

All authors had the same contribution to the article.

References

Barnes

Eveson

Reichart

Sidransky

Pathology and Genetics Head and Neck Tumors: World Health Organization Classification of Tumors. Lyon, France: IARC Press; 2005:254-262.

Riad

Abdel-Rahman

Ezzat

Adly

Dessouky

Shehata

Variables related to recurrence of pleomorphic adenomas: outcome of parotid surgery in 182 cases. Laryngoscope. 2011;121:1467-1472.

Andreasen

Therkildsen

Bjørndal

Homøe

Pleomorphic adenoma of the parotid gland 1985-2010: a Danish nationwide study of incidence, recurrence rate, and malignant transformation. Head Neck. 2016;38:E1364-1369.

Fleming

WB.

Recurrent pleomorphic adenoma of the parotid. Aust N Z J Surg. 1987;57:173-176.

Duck

Mcconnel

FM.

Malignant degeneration of pleomorphic adenoma—clinical implications. Am J Otolaryngol. 1993;14:175-178.

Henriksson

Westrin

Carlsöö

Silfverswärd

Recurrent primary pleomorphic adenomas of salivary gland origin: intrasurgical rupture, histopathologic features, and pseudopodia. Cancer. 1998;1582:617-620.

Hamada

Matsukita

Goto

Mucin expression in pleomorphic adenoma of salivary gland: a potential role for MUC1 as a marker to predict recurrence. J Clin Pathol. 2009;57:813-821.

Soares

Araújo

Juliano

Altemani

Angiogenic and lymphangiogenic microvessel density in recurrent pleomorphic adenoma. J Oral Pathol Med. 2009;38:623-629.

Soares

Altemani

Araújo

VC.

Study of histopathological, morphological and immunohistochemical features of recurrent pleomorphic adenoma: an attempt to predict recurrence of pleomorphic adenoma. J Oral Pathol Med. 2011;40:352-358.

10.

Soares

Demasi

Altemani

de Araújo

VC.

Increased mucin 1 expression in recurrence and malignant transformation of salivary glandpleomorphic adenoma. Histopathology. 2011;58:377-382.

11.

Salzman

Stárek

Kučerová

Skálová

Hoza

Neither expression of VEGF-C/D nor lymph vessel density supports lymphatic invasion as the mechanism responsible for local spread of recurrent salivary pleomorphic adenoma. Virchows Arch. 2014;464:29-34.

12.

Souza

Altemani

Passador-Santos

, et al. Dysregulation of the Rb pathway in recurrent pleomorphic adenoma of the salivary glands. Virchows Arch. 2015;467:295-301.

13.

Zubeldia-Brenner

Roselli

Recabarren

Gonzalez Deniselle

Lara

HE.

Developmental and functional effects of steroid hormones on the neuroendocrine axis and spinal cord. J Neuroendocrinol. 2016;28. doi:10.1111/jne.12401.

14.

Ohshiro

Rayala

Williams

Kumar

El-Naggar

AK.

Biological role of estrogen receptor beta in salivary gland adenocarcinoma cells. Clin Cancer Res. 2006;15:5994-5999.

15.

Glas

Hollema

Nap

Plukker

JT.

Expression of estrogen receptor, progesterone receptor, and insulin-like growth factor receptor-1 and of Mib-1 in patients with recurrent pleomorphic adenoma of the parotid gland. Cancer. 2002;1594:2211-2216.

16.

Alotaibi

Alqarni

Alnobi

Tarakji

Human epidermal growth factor receptor 2 (HER2/Neu) in salivary gland carcinomas: a review of literature. J Clin Diagn Res. 2015;9:04-08.

17.

Etges

Pinto

Kowalski

Soares

Araújo

VC.

Salivary duct carcinoma: immunohistochemical profile of an aggressive salivary gland tumour. J Clin Pathol. 2003;56:914-918.

18.

Falchook

Lippman

Bastida

Kurzrock

Human epidermal receptor 2-amplified salivary duct carcinoma: regression with dual human epidermal receptor 2 inhibition and anti-vascular endothelial growth factor combination treatment. Head Neck. 2014;36:E25-E27.

19.

Rizzardi

Zhang

Vogel

, et al. Quantitative comparison and reproducibility of pathologist scoring and digital image analysis of estrogen receptor B2 immunohistochemistry in prostate cancer. Diagn Pathol. 2016;11:63.

20.

Levy

Elkas

Armstrong

Nieman

LK.

Endometrial effects of prolonged therapy with the selective progesterone receptor modulator ulipristal acetate: a case report. J Reprod Med. 2016;61:159-162.

21.

Lanning

Morrow

Riaz

, et al. The effect of adjuvant trastuzumab on locoregional recurrence of human epidermal growth factor receptor 2-positive breast cancer treated with mastectomy. Ann Surg Oncol. 2015;22:2517-2525.

22.

Gökmen-Polar

Badve

Molecular profiling assays in breast cancer: are we ready for prime time. Oncology. 2012;26:350-357.

23.

Dimery

Jones

Verjan

, et al. Estrogen receptors in normal salivary gland and salivary gland carcinoma. Arch Otolaryngol Head Neck Surg. 1987;113:1082-1085.

24.

Gaffney

Pinkston

Eidson

JJ.

Estrogen receptors in parotid tumors. Endocr Res. 1995;21:635-643.

25.

Kapadia

Barnes

Expression of androgen receptor, gross cystic disease fluid protein, and Cd44 in salivary duct carcinoma. Mod Pathol. 1998;11:1033-1038.

26.

Nasser

Faquin

Dayal

Expression of androgen, estrogen, and progesterone receptors in salivary gland tumors. frequent expression of androgen receptor in a subset of malignant salivary gland tumors. Am J Clin Pathol. 2003;119:801-806.

27.

Di Palma

Skálová

Vanìèek

, et al. Non-invasive (intracapsular) carcinoma ex pleomorphic adenoma: recognition of focal carcinoma by HER-2/Neu and Mib1 immunohistochemistry. Histopathology. 2005;46:144-152.

28.

Fahim

Weinreb

Alexander

, et al. Epithelial proliferation in small ducts of salivary cystadenoma resembling atypical ductal hyperplasia of breast. Head Neck Pathol. 2008;2:213-217.

29.

Barrera

Shroyer

Said

, et al. Estrogen and progesterone receptor and P53 gene expression in adenoid cystic cancer. Head Neck Pathol. 2008;2:13-18.

30.

Ito

Jorge

Vargas

, et al. Histopathological findings of pleomorphic adenomas of the salivary glands. Med Oral Patol Oral Cir Bucal. 2009;114:57-61.

31.

Kolude

Adisa

Adeyemi

Lawal

Immunohistochemical expression of oestrogen receptor-A and progesterone receptor in salivary gland tumours. J Oral Pathol Med. 2013;42:716-719.

32.

Jung

Song

Kim

, et al. Finding and characterizing mammary analogue secretory carcinoma of the salivary gland. Korean J Pathol. 2013;47:36-43.

33.

Locati

Perrone

Losa

, et al. Treatment relevant target immunophenotyping of 139 salivary gland carcinomas. Oral Oncol. 2009;45:986-990.

34.

Hashimoto

Yamamoto

Shiratsuchi

, et al. HER-2/Neu gene amplification in carcinoma ex pleomorphic adenoma in relation to progression and prognosis: a chromogenic in-situ hybridization study. Histopathology. 2012;60:E131-E142.

35.

Sharon

Kelly

Szabo

Sustained response of carcinoma ex pleomorphic adenoma treated with trastuzumab and capecitabine. Head Neck Oncol. 2010;26:2-12.

36.

Laurie

Licitra

Systemic therapy in the palliative management of advanced salivary gland cancers. J Clin Oncol. 2006;24:2673-2678.

37.

Agulnik

Cohen

, et al. Phase II study of lapatinib in recurrent or metastatic epidermal growth factor receptor and/or erbB2 expressing adenoid cystic carcinoma and non adenoid cystic carcinoma malignant tumors of the salivary glands. J Clin Oncol. 2007;25:3978-3984.

38.

Surakanti

Agulnik

Salivary gland malignancies: the role for chemotherapy and molecular targeted agents. Semin Oncol. 2008;35:309-319.

39.

Hammond

Hayes

Wolff

, et al. American society of clinical oncology/college of American pathologists guideline recommendations for immunohistochemical testing of estrogen and progesterone receptors in breast cancer. J Oncol Pract. 2010;6:195-197.

40.

Mouttet

Laé

Caly

, et al. Estrogen-receptor, progesterone-receptor and HER2 status determination in invasive breast cancer. Concordance between immunoistochemistry and MapQuant™ microarray based assay. PLoS ONE. 2016;11:E01464-E01474.

41.

Suzuki

Dobashi

Minato

, et al. EGFR and HER2-Akt-mTOR signaling pathways are activated in subgroups of salivary gland carcinomas. Virchows Arch. 2012;461:271-282.

42.

Ruder

Lubin

Yochanan

, et al. Estrogen and progesterone receptors in breast cancer patients: epidemiologic characteristics and survival differences. Cancer. 1989;64:196-202.

43.

Lamey

Leake

Cowan

, et al. Steroid hormone receptors in human salivary gland tumours. J Clin Pathol. 1987;40:532-534.

44.

Teymoortash

Lippert

Werner

JA.

Steroid hormone receptors in parotid gland cystadenolymphoma (Warthin’s tumour). Clin Otolaryngol Allied Sci. 2001;26:411-416.

45.

Wong

Dobbins

Tseung

, et al. Oestrogen receptor beta expression in pleomorphic adenomas of the parotid gland. J Clin Pathol. 2009;62:789-793.

46.

Glisson

Colevas

Haddad

, et al. HER2 expression in salivary gland carcinomas: dependence on histological subtype. Clin Cancer Res. 2004;110:944-946.

47.

Freitas

Araújo

Martins

Chone

Crespo

Altemani

Biomarker analysis in carcinoma ex pleomorphic adenoma at an early phase of carcinomatous transformation. Int J Surg Pathol. 2005;13:337-342.

48.

Cornolti

Ungari

Morassi

, et al. Amplification and overexpression of HER2/Neu gene and HER2/Neu protein in salivary duct carcinoma of the parotid gland. Arch Otolaryngol Head Neck Surg. 2007;133:1031-1036.

49.

Deroche

Hoschar

Hunt

JL.

Immunohistochemical evaluation of androgen receptor, HER-2/Neu, and P53 in benign pleomorphic adenomas. Arch Pathol Lab Med. 2008;132:1907-1911.

50.

Persson

Andrén

Winnes

, et al. High-resolution genomic profiling of adenomas and carcinomas of the salivary glands reveals amplification, rearrangement, and fusion of Hmga2. Genes Chromosomes Cancer. 2009;48:69-82.

51.

Skálová

Stárek Kucerová

Szépe

, et al. Salivary duct carcinoma—a highly aggressive salivary gland tumor with HER-2/Neu oncoprotein overexpression. Pathol Res Pract. 2001;197:621-626.

52.

Skálová

Stárek

Vanecek

, et al. Expression of HER-2/neu gene and protein in salivary duct carcinomas of parotid gland as revealed by fluorescence in-situ hybridization and Immunohistochemistry. Histopathology. 2003;42:348-356.

53.

Da Cruz Perez

Pires

Alves

, et al. Salivary gland tumors in children and adolescents: a clinicopathologic and immunohistochemical study of fifty-three cases. Int J Pediatr Otorhinolaryngol. 2004;68:895-902.

54.

Rosa

Félix

Fonseca

, et al. Immunoexpression of c-erbB-2 and p53 in benign and malignant salivary neoplasms with myoepithelial differentiation. J Clin Pathol. 1997;50:661-663.

Estrogen Receptor,Progesterone Receptor,and HER-2 Expression in Recurrent Pleomorphic Adenoma

Abstract

Keywords

Introduction

Materials and Methods

Immunohistochemistry

Evaluation of immunohistochemistry staining

CISH assessment of HER-2 status

Statistical analysis

Results

Estrogen receptor

Progesterone receptor

Human epidermal growth factor receptor-2

Chromogenic in situ hybridization

Discussion

Footnotes

Funding:

Declaration of Conflicting Interests:

Author Contributions

References