Correlation of microRNA expression through microarray and their predicted targets with carcinogenesis and prognosis of penile squamous cell carcinoma

INTRODUÇÃO E OBJETIVOS: O câncer de pênis (CaPe) é uma doença rara com alta morbidade e mortalidade. A carcinogênese não é totalmente compreendida e, embora alguns biomarcadores relacionados a prognóstico tenham sido descritos, até o momento nenhum foi adotado na prática clínica. Nosso objetivo foi identificar, através de varredura por microarray, os miRNAs diferencialmente expressos (DEmiR) entre tecido tumoral (TT) versus não neoplásico (TNN) e pacientes com doença metastática versus localizada, bem como identificar os genes diferencialmente expressos (DEG) entre estes grupos e realizar análise integrativa miRNA-mRNA propondo assim elucidar a via de tumorigênese e a descoberta de novos biomarcadores para o CaPe. MÉTODOS: Foram coletadas prospectivamente amostras frescas de TT e TNN adjacente de 24 pacientes com carcinoma espinocelular (CEC) de pênis operados em nossa instituição entre julho de 2015 a janeiro de 2018. Inicialmente, foram selecionados 5 pacientes com doença localizada e 6 com doença metastática linfonodal para realização do microarray. O RNA foi purificado e a análise de varredura com o microarray utilizou o miRNA 4.0 Genechip (Affymetrix). Foram identificados os DEmiRs entre TT e TNN utilizando-se o programa TAC (Affymetrix) com fold change (FC)>2,0, p1,5 e p0,78 (IL1A, MCM2, MMP1, MMP12, SFN e VEGFA). Na análise integrativa, encontramos 8 pares de miRNA-mRNA que mudaram significativamente sua relação, sendo a maior alteração observada para o par miR-432- 5p-TP53. Entre os DEGs da comparação entre os pacientes metastáticos e localizados encontramos 7 DEGs (CCND1, EGFR, ENTPD5, HOXA10, IGF1R, MYC, SNAI2), todos com boa acurácia (AUC>0,74) na distinção entre os grupos. Observamos correlações significativas entre maior expressão do MMP1 com tamanho tumoral (p=0,042), grau (p=0,045), estádio T patológico (p=0,018) e invasão perineural (p=0,007). O gene MMP1 é regulado pelo miR-145-5p que está subexpresso na amostra tumoral. Observamos também correlação entre maior expressão do CCND1 (p=0,006) e EGFR (p=0,001) com grau tumoral; e menor expressão de MMP9 com invasão microvascular (p=0,044). Nas curvas de sobrevida, a maior expressão dos genes CCND1, EGFR, GADD45A, MYC, SNAI2 e SRC e menor dos CDH1, CDKN1A, KLF4 e TP63 estavam associados a mortalidade por CaPe. A maior expressão dos genes FGF2, GADD45A, IL6, MYC, NES e NRP1 e a subexpressão do CDH1 foram correlacionadas a menor sobrevida global. CONCLUSÕES: Descrevemos uma assinatura molecular relacionada à carcinogênese do pênis baseada nas expressões dos miR-432-5p, miR-478b-3p, miR-145-5p, miR-30a-5p, miR-200a-5p, miR-224-5p, miR-31-3p e miR-31-5p e dos genes IL1A, MCM2, MMP1, MMP12, SFN e VEGFA. Identificamos um painel de potenciais biomarcadores de prognóstico no CaPe baseada na expressão dos miR-421, miR-744-5p, miR31-3p e dos genes CCND1, CDH1, CDKN1A, EGFR, ENTPD5, FGF2, GADD45A, HOXA10, IGF1R, KLF4, MMP1, MYC, NES, NRP1, SNAI2, SRC e TP63INTRODUCTION AND OBJECTIVES: Penile cancer (PeCa) is a rare disease with high morbidity and mortality. Its carcinogenesis is not fully understood and, although some prognostic-related biomarkers have been described, none have been adopted in clinical practice to date. Our objective was to identify, through microarray scanning, the differentially expressed miRNAs (DEmiR) between tumor tissue (TT) versus nonneoplastic tissue (NNT) and patients with metastatic versus localized disease, as well as to identify the differentially expressed genes (DEG) between these groups and perform integrative miRNA-mRNA analysis thus proposing to elucidate the tumorigenesis pathway and the discovery of new biomarkers for PeCa. METHODS: Fresh samples of TT and adjacent NNT were prospectively collected from 24 patients with squamous cell carcinoma (SCC) of the penis operated on at our institution between July 2015 and January 2018. Initially, 5 patients with localized disease and 6 with metastatic disease were selected for microarray performance. The RNA was purified and microarray scanning analysis used the miRNA 4.0 Genechip (Affymetrix). The DEmiRs between TT and NNT were identified using the TAC program (Affymetrix) with fold change (FC)>2.0, p1.5 and p0.78 (IL1A, MCM2, MMP1, MMP12, SFN and VEGFA). In the integrative analysis, we found 8 pairs of miRNA-mRNA that significantly changed their relationship, with the greatest change being observed for the pair miR-432-5p-TP53. Among the DEGs in the comparison between metastatic and localized patients, we found 7 DEGs (CCND1, EGFR, ENTPD5, HOXA10, IGF1R, MYC, SNAI2), all with good accuracy (AUC>0.74) in the distinction between groups. We observed significant correlations between increased MMP1 expression and tumor size (p=0.042), grade (p=0.045), pathological T stage (p=0.018) and perineural invasion (p=0.007). The MMP1 gene is regulated by miR-145-5p which is under expressed in the tumor sample. We also observed a correlation between higher expression of CCND1 (p=0.006) and EGFR (p=0.001) with tumor grade; and lower MMP9 expression with microvascular invasion (p=0.044). In the survival curves, higher expression of genes CCND1, EGFR, GADD45A, MYC, SNAI2 and SRC and lower expression of CDH1, CDKN1A, KLF4 and TP63 were associated with mortality from PeCa. Higher expression of FGF2, GADD45A, IL6, MYC, NES and NRP1 genes and under expression of CDH1 were correlated with lower overall survival. CONCLUSIONS: We described a molecular signature related to penile carcinogenesis based on the expressions of miR-432-5p, miR-478b-3p, miR-145-5p, miR-30a-5p, miR-200a-5p, miR-224-5p, miR-31-3p and miR-31-5p and IL1A, MCM2, MMP1, MMP12, SFN and VEGFA genes. We identified a panel of potential prognostic biomarkers in PeCa based on the expression of miR-421, miR-744-5p, miR31-3p and the genes CCND1, CDH1, CDKN1A, EGFR, ENTPD5, FGF2, GADD45A, HOXA10, IGF1R, KLF4, MMP1, MYC, NES, NRP1, SNAI2, SRC and TP6

Comparative study of percutaneous nephrolithotomy performed in the traditional prone position and in three different supine positions

ABSTRACT Purpose: To compare the outcomes of percutaneous nephrolithotomy (PCNL) performed in the prone position (PRON) and in three variations of the supine position. Materials and Methods: We performed a retrospective analysis of patients that underwent PCNL at our institution from June 2011 to October 2016 in PRON and in three variations of the supine position: complete supine (COMPSUP), original Valdivia (VALD), and Galdakao - modified Valdivia (GALD). All patients had a complete pre - operative evaluation, including computed tomography (CT). Success was defined as the absence of residual fragments larger than 4 mm on the first post - operative day CT. Results: We analyzed 393 PCNLs: 100 in COMPSUP, 94 in VALD, 100 in GALD, and 99 in PRON. The overall success rate was 50.9% and was similar among groups (p = 0.428). There were no differences between groups in the number of punctures, stone - free rate, frequency of blood transfusions, drop in hemoglobin level, length of hospital stay, and severe complications (Clavien ≥ 3). COMPSUP had a significantly lower operative time than the other positions. COMPSUP had lower fluoroscopy time than VALD. Conclusion: Patient positioning in PCNL does not seem to impact the rates of success or severe complications. However, COMPSUP is associated with a shorter surgical time than the other positions

Giant Perineal Solitary Fibrous Tumor: A Rare Case Report

Background. Solitary fibrous tumor (SFT) is a fibroblastic mesenchymal tumor that was initially described from the pleura but currently arises at almost every anatomic site. It is usually benign, and surgical resection is curative. SFT involving the perineum is extremely rare. This is the third case report of a perineal SFT in the literature. Case Presentation. We reported an uncommon case of a 64-year-old man presenting with a huge perineal mass that started growing 3 years before his arrival in our service. He was asymptomatic. A contrast-enhanced CT scan revealed a heterogeneous well-circumscribed perineal mass with soft-tissue density. Invasion of the surrounding organs, distal metastasis, and lymph node swelling were absent. The complete resection of mass was done successfully. The specimen was a 23.0 × 14.0 × 8.0 cm encapsulated tumor. Mass weight was 1,170 g. After pathological analysis, we confirmed that the mass was a solitary fibrous tumor. The diagnosis was based on clinical findings and histological morphology and immunohistochemistry study. Conclusion. SFTs are usually indolent tumors with a favorable prognosis. The perineal location is extremely rare. Complete resection of the mass is the treatment of choice

Evaluation of the Incidence of Bladder Perforation After Transurethral Bladder Tumor Resection in a Residency Setting

Purpose: To evaluate prospectively the actual bladder perforation incidence during transurethral resection of bladder tumor (TURB) performed by residents and to identify possible predisposing factors to such condition. Patients and Methods: Thirty-four patients with bladder tumor were submitted to TURB in our academic institution in April 2006, and were prospectively studied. Procedures were all done by senior residents under an attending direct supervision. All patients had a cystograms performed after the procedure by the injection of 400 mL of saline-diluted contrast solution with low-pressure infusion through the Foley catheter. The cystograms were evaluated blindly by a single radiologist. All patients were examined by cystoscopy and/or CT every 3 months for the first 2 years postoperatively. Results: The cystogram showed contrast leaking compatible with bladder perforation in 17 (50%) cases. None of the perforations were recognized intraoperatively by the surgeon. All perforations were extraperitoneal and managed conservatively. There was no significant correlation between the incidence of bladder perforation and the patient age (p = 0.508), the tumor stage (p = 0.998), the tumor grade (p = 0.833), the number of lesions (p = 0.394), and the tumor size (p = 0.651). The only factor that had impact on the development of bladder perforation was tumor localization at the bottom of the bladder (p = 0.035; OR, 6750; 95% CI, 1.14, 39.8). Conclusion: Asymptomatic perforations of the bladder wall occur very frequently after a TURB procedure performed by residents in training and, most of the time, are not noticed by the surgeon. Localization of the tumor at bladder dome was the only factor that negatively influenced perforation rates