Acta Dermato-Venereologica 98-10CompleteContent | Page 19

985 SHORT COMMUNICATION Changes in Androgen Receptor Expression as a Molecular Marker of Progression from Normal Epithelium to Invasive Cancer in Elderly Patients with Penile Squamous Cell Carcinoma Valerio Gaetano VELLONE 1,2# , Emanuele COZZANI 3# , Leonardo PEÑUELA 5# , Roberto RUSSO 3 , Bruno SPINA 2 , Carlo TONCINI 2 , Carlo TERRONE 4# and Aurora PARODI 3# Pathology, Department of Integrated Surgical and Diagnostic Sciences, University of Genoa, 2 Pathology Unit, San Martino Polyclinic Hospital, Section of Dermatology, DISSAL, University of Genoa, San Martino Polyclinic Hospital, Via A. Pastore, 1, IT-16132, Genoa, 4 Department of Urology, University of Genoa, IRCCS San Martino-IST, and 5 Fetal and Perinatal Pathology Unit, Istituto Giannina Gaslini, Genoa, Italy. E-mail: [email protected] # These authors equally contributed to this paper. 1 3 Accepted Jul 5, 2018; Epub ahead of print Jul 6, 2018 Penile squamous cell carcinoma (SCC) is uncommon (1, 2). Two distinct pathways lead to penile SCC: one is linked to human papillomavirus (HPV) infection and is associated with undifferentiated penile intraepithelial neoplasia (PeIN); the other is linked to non-neoplastic epithelial disorders, such as epithelial hyperplasia or li- chen sclerosus (LS) and is associated with differentiated PeIN (3–7). Little is known about molecular changes in the genesis of penile cancer. Only one study has reported the expression of hormonal receptors, such as oestrogen receptors (ERα), and androgen receptor (AR) in normal penile epithelium, with no sign of progesterone receptor (PR) expression (8). Thus far, however, no studies have investigated the possible role of these receptors in the transition from normal epithelium to male genital LS or penile SCC. The aims of the present study were to describe his- tological and immunohistochemical features of SCCs, to examine the modulation of hormonal receptors ERα, AR, PR and molecular proliferation markers (p53 and Ki-67) in normal penile epithelium and in SCC, and to extend this evaluation to penile lesions adjacent to SCC. MATERIALS AND METHODS This retrospective study was conducted on 21 consecutive patients who had undergone surgical resection of penile cancer in San Martino Polyclinic Hospital, Genoa, Italy, in the period 2006 to 2015. All the curative-intended surgical specimens (wide exci- sional biopsies, partial and total penectomies) were fixed in 10% buffered formalin, processed and embedded in paraffin to obtain haematoxylin-eosin-stained slides. In each case, a representative block containing both normal (NE) and tumour tissue (K), as well as the SCC-associated penile lesion (PRE), was chosen in order to obtain further sections. Squamous multi-layered epithelium devoid of major inflammation, and displaying normally orienta- ted cellular maturation, was considered normal epithelium (NE). Squamous cell hyperplasia was defined as epithelial hyperplasia and hyperkeratosis, increased mitotic figures in basal and prickle cell layers and mild dermal chronic inflammation, with no aty- pia. Lichen sclerosus was defined as severe hyperkeratosis, thin epidermis and presence of a homogenized band of dense fibrosis affecting the papillary dermis. Differentiated PeIN was defined as hyperkeratosis, hypergranulosis, acanthosis, with cytological atypia. SCC was defined as variable cytological atypia, high mitotic index, infiltrating pattern and presence of keratin pearls. The following antibodies were used for immunohistochemistry testing: Androgen Receptor (Cell Marque, Rocklin, USA, Clone SP107, pre-diluted), Estrogen Receptor α (Ventana, Innovation Park Dr. Tucson, USA, Clone SP1, pre-diluted), Ki67 (Ventana, Clone 30-9, prediluted), Progesterone Receptor (Ventana, Clone 1E2, pre-diluted), p53 (Ventana, Clone DO-7, prediluted). Appro- priate negative and positive controls were implemented according to the manufacturer’s data-sheets. Expression was evaluated by considering the percentage of cells exhibiting immunoreactivity as well as intracellular localization. In normal tissue and in SCC- associated penile lesions, analysis was restricted to the epithelial component, where the number of positive (brown-stained) cells in a random field of 300 cells was counted; the reactivity pattern of cell positivity was also recorded (i.e. basal, lower, middle and upper thirds of the epithelium, or diffuse). In tumours, the number of positive cells was counted in 5 separate fields of 100 cells each (total number 500 tumour cells). The results were expressed as the mean percentages of positively stained nuclei in relation to the total number of epithelial- or tumour-cell nuclei considered. The staining intensity was not evaluated. RESULTS Clinical-pathological features of the study population are shown in Table SI 1 . Immunohistochemistry Androgen receptor. In NE, SCC-associated lesions and SCCs, AR expression displayed mean  ±  standard deviation (SD) values of 52.0  ±  32.7%, 28.1  ±  28.0% and 0.9  ±  3.0% labelled cells, respectively. Moreover, positivity in SCCs was confined to the lower third of the epithelium; in SCC-associated lesions, the distribution of positivity was variable; in NE, positivity was diffuse. Significant differences (p < 0.001) in staining indexes was observed between NE and K and between PRE and K. Although the AR staining index proved lower in PRE than in NE, the difference failed to reach statistical significance (Fig. S1 1 ). Oestrogen receptors α and progesterone receptor. ERα immunoreactivity was recognized in 5 cases only, and was very weak. None of the specimens tested showed substantial expression of PR. p53. In NE, SCC-associated lesions and SCCs, p53 expression displayed mean  ±  SD values of 2.8  ±  5.0%, https://www.medicaljournals.se/acta/content/abstract/10.2340/00015555-3004 1 This is an open access article under the CC BY-NC license. www.medicaljournals.se/acta Journal Compilation © 2018 Acta Dermato-Venereologica. doi: 10.2340/00015555-3004 Acta Derm Venereol 2018; 98: 985–986