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Additional file 1: Table S1. Genetic characteristics of the HGSOC cells.

Additional file 4: Table S3. Study population of ovarian cancers tested for CCDC6 protein expression.

Additional file 3: Table S2. H-scores of the CCDC6 immunohistochemical staining.

The original version of this Article contained an error in the author affiliations. Silvia Varricchio, Gennaro Ilardi and Stefania Staibanow were incorrectly associated with ‘Department of Public Health, University of Naples "Federico II", Naples, Italy’ instead of the correct ‘Department of Advanced Biomedical Sciences, University of Naples “Federico II”, Naples, Italy.’ This has now been corrected in both the PDF and HTML versions of the Article.

Oral and oropharyngeal cancers represent the two most common malignancies of the head and neck region. The major risk factors for these cancers include alcohol consumption, tobacco use (via smoking or chewing) and high‑risk human papillomavirus infection. The transition from normal epithelium to premalignant tissue and finally carcinoma is in part caused by a summation of genetic and epigenetic modifications. Epigenetic refers to modifications in the way the genome is expressed in cells. The most common examples of epigenetic control of gene expression are DNA methylation, histone modification and regulation by small non‑coding RNAs. The aim of the current paper was to review the recent studies on the main epigenetic changes that have been suggested to serve a role in the carcinogenesis process and progression of oral and oropharyngeal cancers. Furthermore, it is discussed how the epigenetic changes may be used as potential predictive biomarkers and how recent findings in the field may impact the personalized cancer therapy approach for these tumors

Glioblastoma can avoid immune surveillance and induce tumor tolerance, through inhibitory molecules, e.g. PD-L1. Ionizing radiation (IR), used to treat this tumor, is known to increase tumor expression of PD-L1, thus inducing resistance mechanisms. Finding molecular determinants involved in IR-induced PD-L1 may provide a target for preventing such an effect and improve radiotherapy outcomes. We demonstrated that the short isoform of the cochaperone FKBP51 (FKBP51s) regulated PD-L1 expression in melanoma. In glioma, FKBP51s was expressed at high levels, together with PD-L1 and its silencing reduced PD-L1 levels. Conversely, overexpression of FKBP51s increased PD-L1. Different PD-L1 isoforms were observed by immunoblot. A lower band (~37 kDa) corresponding to the naïve protein and two upper bands (~50, ~68 kDa) ascribable to post-translationally modified isoforms. FKBP51s was found mainly bound to the heaviest bands of PD-L1, reasonably mature protein, while the canonical isoform FKBP51 appeared to bind only the naïve protein. Mature PD-L1 protein consists in carbohydrates addition, the principal chemical modification to most plasma membrane proteins, and, particularly, N-glycosylation. Treatment of immunoprecipitated PD-L1 protein with PNGaseF produced a decrease of the highest band and the appearance of a lower band, corresponding to the naïve PD-L1, in accordance with the concept that the heaviest band of PD-L1 is a glycosylated form. Moreover, following subcellular fractionation to obtain extracts from ER and Golgi compartments, we found that naïve 37 kDa PD-L1 was detectable in the ER, but not in the Golgi. The PD-L1 glycosylated band was expressed in ER in a small proportion and mostly in the Golgi. FKBP51s, but not the canonical FKBP51, was found in ER. Co-IP of FKBP51s and PD-L1 from ER extract confirmed the two proteins interacted each other in ER. Our results show that naïve PD-L1 colocalized in the ER of glioma cell complexed with FKBP51s, while the PD-L1 glycosylated form was measured in the Golgi apparatus. Treatment of glioma cell with increasing doses of IR upregulated PD-L1 expression, in a dose-response manner. Particularly, we found a significant increase in PD-L1 expression at 4 and 8 Gy, in comparison with unirradiated glioma cell. Moreover, IR induction of mature PD-L1 was efficiently counteracted by FKBP51s silencing. Subcellular fractionation of glioma cell subjected to IR in kinetics showed an early and transitory decrease in FKBP51s ER levels at 3hrs, in line with a reduction of the glycosylated band in the whole lysate. After 8 hrs from IR, FKBP51s rose up again in the ER inducing a full maturation of PD-L1. These findings suggested that FKBP51s has a role in catalyzing PD-L1 folding, an essential step to glycosylation, through which it controls the affinity for PD1. This study identifies FKBP51s as an essential element that regulates PD-L1 expression on glioma cell, which is exploited by the tumor to resist to IR. Citation Format: Paolo D'Arrigo, Michele Russo, Elia Guadagno, Roberto Pacelli, Maria Laura Del Basso De caro, Anna Rea, Martina Tufano, Stefania Staibano, Gennaro Ilardi, Maria Fiammetta Romano, Simona Romano. Ionizing radiation-induced PD-L1 upregulation in glioma: a crucial role for the molecular chaperone FKBP5 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 586. doi:10.1158/1538-7445.AM2017-586

Melanoma delivers immune suppressive stimuli through PDL-1. FK506 binding protein 51 (FKBP51) is an immunophilin capable of immune suppression. We, previously, demonstrated a relevant role for this protein in melanoma biology and progression. Melanoma also expresses a splicing variant of FKBP51 (isoform 2 or ISO2), whose function is unknown. Aim of this study was to generate knowledge on the mechanism regulating PDL- 1 expression in melanoma, and find biomarkers predictive of response to immunotherapy.

Polyclonal antibodies against somatostatin receptors (SSTRs) available up to now recognizing intracellular sites of receptors and their recycling products do not detect bioactive ligand binding domains (LBDs) and are of limited performance in paraffin-embedded tissues. Aim of this study was to evaluate by immunohistochemistry the expression of SSTR5 on an archival series of pituitary tumors using a new MoAbs against the SST-binding domain (Y-SSTR5). Methods: We used paraffined tissue sections from 20 non-secreting pituitary tumors (NS) and 14 PRL-secreting (PRL) adenomas and, as control, sections from normal tissue surrounding the same tumor. A standard streptavidin-biotin-labeled peroxidase immunostaining was performed. Negative controls were performed with non immune serum. The degree of immunopositivity was evaluated semi-quantitatively according to an arbitrary scale. Results: Immunostaining for SSTR5 was found in 90% of NS (low-medium 55%, high 35%) and in 100% of PRLS adenomas (low-medium 14%, high 86%). Conclusion: SSTR5 was found in the majority of pituitary tumors examined. PRLS adenomas showed a tendency to higher expression of receptor protein. Y SSTR5 MoAb can be used to detect SSTR5 on the membrane of pituitary cells in classical paraffin embedded histological sections. The specific detection of LBD SSTR5 by MoAb can support a potential choice of panligand or SST analogues targeting SSTR5 in resistant/recurrent pituitary tumors.

Very recently, we have identified FKBP51, an immunophilin phisiologically expressed in lymphocytes, as a protein associated with malignant melanoma. Its expression correlates with tumor aggressiveness and is maximal in metastatic lesions. We have also demonstrated that FKBP51 is a factor of resistance to genotoxic agents because it promotes NF-κB activation. Aim of the present study was to investigate FKBP51 expression in solid tumors. Overall 71 samples, stored in our Patology section, of the following tumors: breast, colon, lung, pancreas, ovary and prostate (10 for each tumor, with exception of colon for which 21 samples were analyzed), and a comparable number of normal tissue samples, were analysed for FKBP51 expression, by immunohistochemistry. An intense signal was observed in all prostatic, ovarian and lung adenocarcinomas analyzed and in 8 out of 10 pancreatic cancers. Immunohistochemistry was low positive/negative in breast cancer. Normal tissues of the same histotype showed very low or undetectable levels of FKBP51, except for colon. FKBP51 was expressed in normal colonic mucosa, particularly in proliferative basal zone of normal glands. Nevertheless, an increased expression was found in 15 out of 21 tumor analyzed. Interestingly, we found that in colon carcinoma FKBP51 localization changed with grading. The protein was mostly cytosolic in G1, whereas it was always nuclear in G3. A nuclear localization was also observed in lung cancer. We also measured FKBP51 mRNA levels in deparaffinized tissues both normal and tumoral, by Realtime PCR. All normal tissues, except for prostate, displayed mRNA levels lower than those of PBL. FKBP51 levels in prostate and PBL were similar. Increases in mRNA levels, from 3 to 30 folds, were found in prostate, ovary, colon and lung cancers, compared to normal tissue of the same histotype. A correlation of gene expression with grading has been found in pancreatic adenocarcinoma. In undifferentiated carcinoma, an increase of more than 200fold of FKBP51 mRNA level was found, compared to normal tissue, whereas a moderate 2-4 fold increase was found in G2 tumors and no increase in G1. Very low levels were found in breast cancer, although higher than in normal tissue. In conclusion, our results show that FKBP51 expression increases in different solid tumors in comparison with the normal tissue counterpart. This study supports the conclusion that deregulated FKBP51 plays a role in malignant transformation.

Poly(ADP-ribosylation) is a major post-translational (epigenetic) modification involving a large number of proteins of eukaryotic cells. Several lines of evidence indicate that the Poly(ADP-ribosylation) of histones, beyond the well-known activity in DNA strand breaks repair, plays a pivotal role also in chromatin structure, replication and assembly [1–2].A critical role in chromosome assembly and replication has been shown also for the Chromatin Assembly Factor (CAF)-1 complex, which expresses at high level its p60 subunit whenever a hyperproliferative condition takes place in eukaryotic cells [3–4].We examined the immunohistochemical expression of PARP-1 and CAF-1/p60 in a selected series of squamous cell carcinomas (SCC) of the head and neck region, to evaluate its relationship with the clinical and pathological features of tumors.The results of the study support the existence of a statistically significative correlation between the overexpression of both the proteins and the worse prognosis of patients, outlining the role of epigenetic events in the gain of aggressiveness of skin and mucosal SCC, and the role of PARP-1 and CAF-1/p60 as specific promising targets for molecular therapies for SCC.

BACKGROUND: Cutaneous melanoma (CM) is the most lethal form of skin malignancy, which registers a constant increase in incidence worldwide. The identification of molecular alteration(s) involved in its biological aggressiveness represents a major challenge for researchers, considering that existing therapies are ineffective to treat metastasizing cases. The epigenetic control of chromatin dynamics during DNA synthesis, replication, and repair is fundamental for the orderly progression of cell proliferation. The Chromatin Assembly Factor 1 (CAF-1) complex acts as a major regulator of this process; its intermediate (p60) subunit has been recently proposed as a novel proliferation and prognostic marker for several tumors. We aimed to establish if the evaluation of the expression of CAF-1/p60 in primary CM may help define the prevision of outcome of patients. METHODS: Immunohistochemistry with anti-CAF-1/p60 was performed on paraffin-embedded tissue sections of 130 cases of primary CM retrieved from the archive files of the Department of Biomorphological and Functional Sciences, Section of Pathology, University "Federico II" of Naples, Italy. Results were compared with histopathological and follow-up data of patients. RESULTS: CAF-1/p60 was expressed in all CM. A significant statistical association between the overexpression of the protein and the occurrence of skin, node and/or distant metastases (P < 0.05) emerged, independently from histopathological prognostic factors. CONCLUSIONS: CAF-1/p60 looks promising as a new prognostic marker for CM and sheds new light on the molecular events associated with photocancerogenesis and melanoma biology.The screening for CAF-1/p60 might contribute to the molecular sub-classification of CM, with improved translational outcomes.

Cutaneous melanoma (CM) is the most lethal skin malignancy, with a constant increase in incidence. These tumors show a biological behavior unpredictable by the classical parameters Moreover, conventional therapies are ineffective to treat advanced stage case. Therefore, the identification of molecular alterations involved in their biological aggressiveness represents a major challenge for researchers. Accumulating evidence supports the involvement of cancer stem cells (CSC) in initiation, progression, chemoresistance and therapeutic failure of malignant melanoma, and the aggressive subsets of melanoma cells have been frequently associated with molecular markers shared by stem cells.

Il carcinoma squamoso del cavo orale (OSCC) è il tumore più frequente della regione testa-collo. A livello mondiale, rappresenta il quinto tumore per incidenza e la sesta causa di mortalità per cancro negli Stati Uniti. La chirurgia rappresenta lo standard terapeutico per questo tumore, nelle fasi iniziali di sviluppo. Tuttavia, nelle fasi avanzate di progressione, gli OSCC sono non-responsivi a radio/chemioterapia, e frequentemente mostrano un decorso inesorabilmente fatale. I meccanismi biomolecolari responsabili della scarsa risposta degli OSCC alla terapia restano tuttora in gran parte sconosciuti. Recentemente, è stato evidenziato che un sottogruppo di OSCC, positivi per infezione da HPV, presentano un comportamento meno aggressivo ed una migliore risposta alla radioterapia. Abbiamo quindi studiato una serie di OSCC primitivi e metastatici, con o senza associata infezione da HPV, confrontandone i dati di follow-up con l’espressione immunoistochimica del fattore 1 di assemblaggio cromatinico (CAF1) e di marcatori di staminalità. E’ infatti stato evidenziato recentemente che CAF-1, istone chaperone con ruolo altamente specializzato durante la replicazione e riparazione del DNA, costituisce un sensibile marker di proliferazione, di valore prognostico in diverse neoplasie umane, comprendenti l’OSCC. Inoltre, si moltiplicano in letteratura le segnalazioni di un ruolo rilevante della staminalità nell’acquisizione dell’aggressività biologica dei tumori maligni. I nostri risultati indicano che la simultanea espressione di CAF1 e di markers di staminalità si associa al gruppo di OSCC metastatizzanti, ed è inversamente proporzionale alla presenza dell’HPV. Ciò potrà rivelarsi utile nella selezione del sottogruppo di pazienti con OSCC da candidare a nuove terapie molecolari con bersaglio costituito da marcatori di staminalità e/o proteine del complesso CAF-1.

Basal cell carcinoma (BCC) is the most common cancer in the white-skinned population accounting for about 15% of all neoplasms. Its incidence is increasing worldwide, at a rate of about 10% per year. BCC, although infrequently metastasizing, very often causes extensive tissue losses, due to the high propensity toward stromal infiltration, particularly in its dedifferentiated forms, with disfiguring and debilitating results. To date, there still is limited availability of therapeutic treatments alternative to surgery. We evaluated the immunohistochemical expression of the carbonic anhydrase IX (CAIX), one of the main markers of tissue hypoxia, in a set of 85 archived FFPE BCC tissues, including the main subtypes, with different clinical outcomes, to demonstrate a possible relationship between hypoxic phenotype and biological aggressiveness of these neoplasms. Our results showed that the expression level of the CAIX protein contributes to the stratification of BCC in the different risk classes for recurrence. We hypothesize for CAIX a potential therapeutic role as a target therapy in the treatment of more aggressive BCCs, thus providing an alternative to surgical and pharmacological therapy with Hedgehog inhibitors, a promising example of target therapy in BCCs.

Uveal melanoma (UM) is the most frequent primary ocular malignancy of adults; it exhibits an almost invariably poor prognosis with onset of liver metastases within 10–15 years after the diagnosis. Serine and arginine-rich splicing factor 1 (SRSF1) is an RNA-binding protein with proto-oncogene functions, including stimulation of angiogenesis, cell migration and cell growth; regarding the complex regulation of tumor angiogenesis, it has been suggested that SRSF1 regulates the alternative splicing of vascular endothelial growth factor-α, promoting the formation of its pro-angiogenic isoform. The immunohistochemical expression of SRSF1 on a series of 85 primary UMs, including 39 metastasizing and 46 non-metastasizing cases, was investigated; to clarify the potential pathogenetic role of SRSF1 in this tumor and its effect on angiogenesis, we correlated our immunohistochemical findings with the clinico-pathological features, the prognostic data and blood vascular microvessel density (MVD) findings of the cases from our series. Cases with higher immunohistochemical expression of SRSF1 also had higher MVD, higher metastatic potential and shorter metastasis-free survival; conversely, cases with lower SRSF1 immunoexpression showed lower MVD, lower metastatic risk and longer metastasis-free survival times. Our results suggested that SRSF1 has a negative prognostic role and a pro-angiogenic function in UM.

Early events of basal cell carcinoma (BCC) tumorigenesis are triggered by inappropriate activation of SHH signaling, via the loss of Patched1 (Ptch1) or by activating mutations of Smoothened (Smo). TBX1 is a key regulator of pharyngeal development, mainly through expression in multipotent progenitor cells of the cardiopharyngeal lineage. This transcription factor is connected to several major signaling systems, such as FGF, WNT, and SHH, and it has been linked to cell proliferation and to the regulation of cell shape and cell dynamics. Here, we show that TBX1 was expressed in all of the 51 BCC samples that we have tested, while in healthy human skin it was only expressed in the hair follicle. Signal intensity and distribution was heterogeneous among tumor samples. Experiments performed on a cellular model of mouse BCC showed that Tbx1 is downstream to GLI2, a factor in the SHH signaling, and that, in turn, it regulates the expression of Dvl2, which encodes an adaptor protein that is necessary for the transduction of WNT signaling. Consistently, Tbx1 depletion in the cellular model significantly reduced cell migration. These results suggest that TBX1 is part of a core transcription network that promotes BCC tumorigenesis.

Purpose: To describe functional and histopathological findings after macular peeling using different dyes. Methods: Prospective, randomized, comparative, interventional, and immunohistochemical study. Forty-five eyes from 45 patients with idiopathic epiretinal membrane (ERM) underwent pars plana chromovitrectomy with ERM and inner limiting membrane (ILM) using trypan blue 0.15% + brilliant blue 0.05% + lutein 2% in group 1 (15 eyes), trypan blue 0.15% + brilliant blue 0.025% + polyethylene glycol 3350 4% in group 2 (15 eyes), and indocyanine green 0.05% in group 3 (15 eyes). We evaluated visual acuity (VA) and macular sensitivity (MS) preoperatively, 1, 3, and 6 months after surgery. The expression of glial fibrillary acidic protein (GFAP) and neurofilament protein (NF) was assessed immunohistochemically on the ILMs peeled as markers of glial and neuronal cells. Results: In group 1, both mean VA and MS were significantly better at 1 and 3 months after surgery (P < 0.05), whereas no significant difference was found after 6 months. GFAP and NF expression was significantly lower in group 1 (P < 0.05). Conclusions: The ERM/ILM peeling is thought to rip off the intraretinal tissue, based on the amounts of GFAP and NF in the specimens. The use of lutein dyes reduces iatrogenic stress to the retinal tissue and allows a faster functional recovery in the first 3 months after surgery, suggesting a less iatrogenic adhesion to the retinal tissue

Background The muscle invasive form of urothelial bladder cancer (UBC) is a deadly disease. Currently, the therapeutic approach of UBC is mostly based on surgery and standard chemotherapy. Biomarkers to establish appropriate drugs usage are missing. Deficiency of the tumor suppressor CCDC6 determines PARP-inhibitor sensitivity. The CCDC6 levels are modulated by the deubiquitinase USP7. In this work we scored CCDC6 and USP7 expression levels in primary UBC and we evaluated the expression levels of CCDC6 in correlation with the effects of the PARP-inhibitors combined with the USP7 inhibitor, P5091, in vitro. Since PARP-inhibitors could be enhanced by conventional chemotherapy or DNA damage inducers, we tested the new agent RRx-001, able to induce DNA damage, to prove the benefit of combined treatments in bladder cancer cells. Methods The J82, T24, 5637 and KU-19-19 bladder cancer cells were exposed to USP7 inhibitor P5091 in presence of cycloheximide to analyse the CCDC6 stability. Upon the CCDC6 degradation induced by P5091, the cells sensitivity to PARP-inhibitor was evaluated by cell viability assays. The ability of the DNA damage inducer RRx-001 to modulate CCDC6 protein levels and H2AX phosphorylation was detected at immunoblot. The combination of USP7 inhibitor plus RRx-001 enhanced the PARP-inhibitor sensitivity, as evaluated by cell viability assays. The results of the scores and correlation of CCDC6 and USP7 expression levels obtained by UBC primary biopsies staining were used to cluster patients by a K-mean cluster analysis. Results P5091 determining CCDC6 degradation promoted bladder cancer cells sensitivity to PARP-inhibitor drugs. RRx-001, by inducing DNA damage, enhanced the effects of the combined treatment. The immunohistochemical staining of both CCDC6 and USP7 proteins allowed to cluster the high grade (G3) UBC patients, on the basis of CCDC6 expression levels. Conclusions In high grade UBC the identification of two clusters of patients based on CCDC6 and USP7 expession can possibly indicate the use of PARP-inhibitor drugs, in combination with USP7 inhibitor in addition to the DNA damage inducer RRx-001, that also acts as an immunomodulatory agent, offering novel therapeutic strategy for personalized medicine in bladder cancer patients. Electronic supplementary material The online version of this article (10.1186/s13046-019-1087-1) contains supplementary material, which is available to authorized users.

Tumor necrosis factor-α (TNF-α) is a pleiotropic cytokine, whose role in melanoma is controversial. Although high-dose TNF-α is approved for the treatment of patients with in transit-metastatic melanoma confined to the limb, diverse preclinical models of melanoma have shown that TNF-α can induce cell invasion. Biomarkers that can differentiate between the dual role of TNF-α are needed. TRAF2 is critical to TNF receptor-induced activation of nuclear factor-κB (NF-κB), allowing shifting from death to survival-signaling cascades. The large immunophilin FKBP51 acts as a scaffold and catalyst in the IκB kinase complex assembly and activation. Here, using microscopy and an electrophoretic mobility-shift assay, we provide further evidence in support of the essential role of FKBP51 in sustaining the TNF-α NF-κB signaling in melanoma. Through the cross-linking reaction with the chemical linker disuccinimidyl glutarate, we show that a direct interaction occurs between FKBP51 and TRAF2 in melanoma cells. Immunohistochemistry of tumor samples from 24 patients with cutaneous melanomas showed a correlation between the expressions of the two proteins. Given the association of FKBP51 and TRAF2 with TNF-α-induced NF-κB signaling and their correlation in tumor samples, we propose that the two proteins can be exploited as useful markers for the identification of those melanoma tumors that can benefit from TNF-α inhibition. Future studies will address this hypothesis.

Germ cell tumors (GCT) are the most common malignancies in males between 15 and 35 years of age. Despite the high cure rate, achieved through chemotherapy and/or surgery, the molecular basis of GCT etiology is still largely obscure. Here, we show a positive correlation between MAPK15 (ERK8; ERK7) expression and specific GCT subtypes, with the highest levels found in the aggressive embryonal carcinomas (EC). Indeed, in corresponding cellular models for EC, MAPK15 enhanced tumorigenicity in vivo and promoted cell proliferation in vitro, supporting a role for this kinase in human GCT. At molecular level, we demonstrated that endogenous MAPK15 is necessary to sustain cell cycle progression of EC cells, by limiting p53 activation and preventing the triggering of p53-dependent mechanisms resulting in cell cycle arrest. To understand MAPK15-dependent mechanisms impinging on p53 activation, we demonstrate that this kinase efficiently protects cells from DNA damage. Moreover, we show that the ability of MAPK15 to control the autophagic process is necessary for basal management of DNA damage and for tumor formation controlled by the kinase. In conclusion, our findings suggest that MAPK15 overexpression may contribute to the malignant transformation of germ cells by controlling a "stress support" autophagic pathway, able to prevent DNA damage and the consequent activation of the p53 tumor suppressor. Moreover, in light of these results, MAPK15-specific inhibitors might represent new tools to enhance the therapeutic index of cytotoxic therapy in GCT treatment, and to increase the sensitivity to DNA-damaging drugs in other chemotherapy-resistant human tumors.

Systemic sclerosis (SSc) is a chronic autoimmune disease characterized by fibrosis, alteration in the microvasculature and immunologic abnormalities. It has been hypothesized that an abnormal redox state could regulate the persistent fibrotic phenotype in SSc patients. N-Formyl peptide receptors (FPRs) are chemotactic receptors overexpressed in fibroblasts derived from SSc patients. In this study, we demonstrated that stimulation of FPRs promotes the generation of reactive oxygen species (ROS) in skin fibroblasts. In fibroblast cells, ROS production was due to FPRs interaction with the urokinase receptor (uPAR) and to β1 integrin engagement. FPRs cross-talk with uPAR and integrins led to Rac1 and ERKs activation. FPRs stimulation increased gp91phox and p67phox expression as well as the direct interaction between GTP-Rac1 and p67phox, thus promoting assembly and activation of the NADPH oxidase complex. FPRs functions occur through interaction with a specific domain of uPAR (residues 88SRSRY92) that can be exposed on the cell membrane by protease-mediated receptor cleavage. Immunohistochemistry analysis with a specific anti-SRSRY antibody showed increased expression of uPAR in a cleaved form, which exposes the SRSRY sequence at its N-terminus (DIIDIII-uPAR88–92) in skin biopsies from SSc patients. As expected by the increased expression of both FPRs and DII-DIII-uPAR88-92, fibroblasts derived from SSc patients showed a significantly increase in ROS generation both at a basal level than after FPRs stimulation, as compared to fibroblasts from normal subjects. C37, a small molecule blocking the interaction between FPRs and uPAR, and selumetinib, a clinically approved MAPKK/ERK inhibitor, significantly inhibited FPRs-mediated ROS production in fibroblasts derived from SSc patients. Thus, FPRs, through the interaction with the uPA/uPAR system, can induce ROS generation in fibroblasts by activating the NADPH oxidase, playing a role in the alteration of the redox state observed in SSc.

Basal-cell carcinoma (BCC) is the most common non-melanoma skin cancer (NMSC) in white individuals. The main risk factor of BCC is intense exposure to ultraviolet radiation. The gold standard of diagnosis of BCC is histopathology. Treatment options for BCC consist of surgery, vismodegib, radiotherapy and topical or ablative treatments. Surgical excision is usually the preferred treatment for nodular and aggressive BCC subtypes. Here in, we report the case of a 51 year-old Caucasian woman successfully treated with ingenol mebutate (IM) 0.05% gel on a large-sized (ø3 cm) superficial BCC (sBCC) localized on the right shoulder. IM appears to be an effective, simple, safe and comfortable treatment even for large-sized sBCC with good cosmetic results.

Human Papilloma Virus (HPV) can play a causative role in the development of sinonasal tract malignancies. In fact, HPV may be the most significant causative agent implicated in sinonasal tumorigenesis and is implicated in as many as 21% of sinonasal carcinomas. To date, there are no definitive, reliable and cost-effective, diagnostic tests approved by the FDA for the unequivocal determination of HPV status in head and neck cancers. We followed an exhaustive algorithm to correctly test HPV infection, including a sequential approach with p16INK4a IHC, viral DNA genotyping and in situ hybridization for E6/E7 mRNA. Here, we report a case of sinonasal carcinoma with discordant results using HPV test assays. The tumor we describe showed an irregular immunoreactivity for p16INK4a, and it tested positive for HPV DNA; nevertheless, it was negative for HR-HPV mRNA. We discuss the possible meaning of this discrepancy. It would be advisable to test HPV transcriptional status of sinonasal carcinoma on a diagnostic routine basis, not only by p16INK4a IHC assay, but also by HPV DNA genotyping and HR-HPV mRNA assessment.

Introduction Epiretinal traction is not responsible only for epiretinal but also intraretinal changes. This study aims to describe structural and vascular intraretinal changes after macular peeling in idiopathic (iERM) vs diabetic ERM (dERM). Methods We conducted a prospective interventional study on forty-two eyes, 23 with iERMs and 19 with dERMs, undergoing ERM-ILM peeling. We performed SD-OCT preoperatively, 1 and 6 months postoperatively to assess central macular thickness (CMT), intraretinal cysts (IC) and/or continuous ectopic inner foveal layers (CEIFL), superficial and deep capillary free zone (CFZ) area on OCT-A. Glial fibrillary acidic protein (GFAP), as a Muller cells marker, was detected immunohistochemically on ILM specimens, to assess Muller cells iatrogenic damage. Results The CEIFLs were significantly more common in iERMs (12 (52.2%) in iERMs vs 2 (10.5%) in dERMs, p = 0.004), whereas ICs in dERMs (6 (26.1%) in iERMs vs 17 (89.5%) in dERMs, p

Granular cell tumor (GCT), a relatively rare neuroectodermal tumor occurring most often in the head and neck region, is not uncommonly associated with pseudoepitheliomatous hyperplasia of the overlying surface epithelium, which may be at times nonreadily distinguishable from well-differentiated squamous cell carcinoma (SCC). To the best of our knowledge, only a handful of coexisting SCC and GCT, mostly described in the esophagus, have been reported in (the current) literature so far. We herein report 2 new cases of coexisting GCT and SCC of the head and neck region, located, respectively, in larynx and tongue; comment on their clinical, imaging, and pathologic features; and discuss their management. In the present work, we also review the literature concerning this association to contribute to the head and neck pathologists’ and surgeons’ awareness regarding the possibility of this association for an adequate surgical excision and a better management of these patients.

// Francesco Morra 1 , Francesco Merolla 2, 3 , Virginia Napolitano 1, 2 , Gennaro Ilardi 2 , Caterina Miro 1 , Simona Paladino 4 , Stefania Staibano 2 , Aniello Cerrato 1 , Angela Celetti 1 1 Institute for Experimental Endocrinology and Oncology, Research National Council, Naples, Italy 2 Department of Advanced Biomedical Sciences, University “Federico II”, Naples, Italy 3 Department of Medicine and Health Sciences “V. Tiberio”, University of Molise, Campobasso, Italy 4 Department of Molecular Medicine and Medical Biotechnology, University “Federico II”, Naples, Italy Correspondence to: Angela Celetti, email: celetti@unina.it Keywords: CCDC6, USP7, ARFL and V7, P5091, olaparib Received: December 08, 2016 Accepted: March 15, 2017 Published: March 22, 2017 ABSTRACT Purpose of the study: Reduced levels of the tumor suppressor protein CCDC6 sensitize cancer cells to the treatment with PARP-inhibitors. The turnover of CCDC6 protein is regulated by the de-ubiquitinase USP7, which also controls the androgen receptor (AR) stability. Here, we correlated the expression levels of CCDC6 and USP7 proteins in primary prostate cancers (PC). Moreover, we tested the efficacy of the USP7 inhibitors, in combination with PARP-inhibitors as a novel therapeutic option in advanced prostate cancer. Experimental techniques: PC cells were exposed to USP7 inhibitor, P5091, together with cycloheximide, to investigate the turnover of the USP7 substrates, AR and CCDC6. As outcome of the AR downregulation, transcription targets of AR and its variant V7 were examined by qPCR. As a result of CCDC6 degradation, the induction of PARP inhibitors sensitivity was evaluated by analyzing PC cells viability and foci formation. We scored and correlated CCDC6 and USP7 expression levels in a prostate cancer tissue microarray (TMA). Results: P5091 accelerated the degradation of AR and V7 isoform affecting PSA, UBE2C, CDC20 transcription and PC cells proliferation. Moreover, P5091 accelerated the degradation of CCDC6 sensitizing the cells to PARP-inhibitors, that acted sinergistically with genotoxic agents. The immunohistochemical analysis of both CCDC6 and USP7 proteins exhibited significant correlation for the intensity of staining ( p ≤ 0.05). Data interpretation: Thus, CCDC6 and USP7 represent predictive markers for the combined treatment of the USP7-inhibitors and PARP-inhibitors in advanced prostate cancer.