C-Met/miR-130b axis as novel mechanism and biomarker for castration resistance state acquisition.

Although a significant subset of prostate tumors remain indolent during the entire life, the advanced forms are still one of the leading cause of cancer-related death. There are not reliable markers distinguishing indolent from aggressive forms. Here we highlighted a new molecular circuitry involving microRNA and coding genes promoting cancer progression and castration resistance. Our preclinical and clinical data demonstrated that c-Met activation increases miR-130b levels, inhibits androgen receptor expression, promotes cancer spreading and resistance to hormone ablation therapy. The relevance of these findings was confirmed on patients' samples and by in silico analysis on an independent patient cohort from Taylor's platform. Data suggest c-Met/miR-130b axis as a new prognostic marker for patients' risk assessment and as an indicator of therapy resistance. Our results propose new biomarkers for therapy decision-making in all phases of the pathology. Data may help identify high-risk patients to be treated with adjuvant therapy together with alternative cure for castration-resistant forms while facilitating the identification of possible patients candidates for anti-Met therapy. In addition, we demonstrated that it is possible to evaluate Met/miR-130b axis expression in exosomes isolated from peripheral blood of surgery candidates and advanced patients offering a new non-invasive tool for active surveillance and therapy monitoring.

Papillary type 2 versus clear cell renal cell carcinoma: Survival outcomes.

AIM: To compare the cancer specific survival (CSS) between p2-RCC and a Propensity Score Matched (PSM) cohort of cc-RCC patients. METHODS: Fifty-five (4.6%) patients with p2-RCC and 920 cc-RCC patients were identified within a prospectively maintained institutional dataset of 1205 histologically proved RCC patients treated with either RN or PN. Univariable and multivariable Cox regression analyses were used to identify predictors of CSS after surgical treatment. A 1:2 PSM analysis based on independent predictors of oncologic outcomes was employed and CSS was compared between PSM selected cc-RCC patients using Kaplan-Meier and Cox regression analysis. RESULTS: Overall, 55 (4.6%) p2-RCC and 920 (76.3%) cc-RCC patients were selected from the database; p2-RCC were significantly larger (p = 0.001), more frequently locally advanced (p < 0.001) and node positive (p < 0.001) and had significantly higher Fuhrman grade (p < 0.001) than cc-RCC. On multivariable Cox regression analysis age (p = 0.025), histologic subtype (p = 0.029), pN stage (p = 0.006), size, pT stage, cM stage, sarcomatoid features and Fuhrman grade (all p < 0.001) were independent predictors of CSS. After applying the PSM, 82 cc-RCC selected cases were comparable to 41 p2-RCC for age (p = 0.81), tumor size (p = 0.39), pT (p = 1.00) and pN (p = 0.62) stages, cM stage (p = 0.71) and Fuhrman grade (p = 1). In this PSM cohort, 5 yr CSS was significantly lower in the p2-RCC (63% vs 72.4%; p = 0.047). At multivariable Cox analysis p2 histology was an independent predictor of CSM (HR 2.46, 95% CI 1.04-5.83; p = 0.041). CONCLUSIONS: We confirmed the tendency of p2-RCC to present as locally advanced and metastatic disease more frequently than cc-RCC and demonstrated p2-RCC histology as an independent predictor of worse oncologic outcomes.

A microRNA code for prostate cancer metastasis.

Although the development of bone metastasis is a major detrimental event in prostate cancer, the molecular mechanisms responsible for bone homing and destruction remain largely unknown. Here we show that loss of miR-15 and miR-16 in cooperation with increased miR-21 expression promote prostate cancer spreading and bone lesions. This combination of microRNA endows bone-metastatic potential to prostate cancer cells. Concomitant loss of miR-15/miR-16 and gain of miR-21 aberrantly activate TGF-ß and Hedgehog signaling, that mediate local invasion, distant bone marrow colonization and osteolysis by prostate cancer cells. These findings establish a new molecular circuitry for prostate cancer metastasis that was validated in patients' cohorts. Our data indicate a network of biomarkers and druggable pathways to improve patient treatment.

Silencing of GSTP1, a prostate cancer prognostic gene, by the estrogen receptor-ß and endothelial nitric oxide synthase complex.

We recently identified in prostate tumors (PCa) a transcriptional prognostic signature comprising a significant number of genes differentially regulated in patients with worse clinical outcome. Induction of up-regulated genes was due to chromatin remodeling by a combinatorial complex between estrogen receptor (ER)-ß and endothelial nitric oxide synthase (eNOS). Here we show that this complex can also repress transcription of prognostic genes that are down-regulated in PCa, such as the glutathione transferase gene GSTP1. Silencing of GSTP1 is a common early event in prostate carcinogenesis, frequently caused by promoter hypermethylation. We validated loss of glutathione transferase (GST) P1-1 expression in vivo, in tissue microarrays from a retrospective cohort of patients, and correlated it with decreased disease-specific survival. Furthermore, we show that in PCa cultured cells ERß/eNOS causes GSTP1 repression by being recruited at estrogen responsive elements in the gene promoter with consequential remodeling of local chromatin. Treatment with ERß antagonist or its natural ligand 5α-androstane-3ß,17ß-diol, eNOS inhibitors or ERß small interference RNA abrogated the binding and reversed GSTP1 silencing, demonstrating the direct involvement of the complex. In vitro, GSTP1 silencing by ERß/eNOS was specific for cells from patients with worse clinical outcome where it appeared the sole mechanism regulating GSTP1 expression because no promoter hypermethylation was present. However, in vivo chromatin immunoprecipitation assays on fresh PCa tissues demonstrated that silencing by ERß/eNOS can coexist with promoter hypermethylation. Our findings reveal that the ERß/eNOS complex can exert transcriptional repression and suggest that this may represent an epigenetic event favoring inactivation of the GSTP1 locus by methylation. Moreover, abrogation of ERß/eNOS function by 3ß-adiol emphasizes the significance of circulating or locally produced sex steroid hormones or their metabolites in PCa biology with relevant clinical/therapeutic implications.

Deregulation of Aurora kinase gene expression in human testicular germ cell tumours.

The Aurora kinases regulate chromosome segregation and cytokinesis, and alterations in their expression associate with cell malignant transformation. In this study, we demonstrated by qRT-PCR analysis of 14 seminomas that Aurora-A mRNA was, with respect to control tissues, augmented in five of 14 tumour tissues by 2.17 +/- 0.30 fold (P < 0.05) and reduced in 9 to 0.38 +/- 0.10 (P < 0.01). Aurora-B mRNA was increased in 11 tumour tissues by 4.33 +/- 0.82 fold (P < 0.01) and reduced in 3 to 0.41 +/- 0.11 fold. Aurora-C mRNA was reduced to 0.20 +/- 0.32 fold (P < 0.01) in 13 seminomas and up-regulated in one case. Western blot experiments, performed on protein extracts of nine seminomas and six normal testes, showed an up-regulation of Aurora-B protein by 10.14 +/- 3.51 fold (P < 0.05), while Aurora-A protein was found increased in four seminomas by 2.16 +/- 0.43 (P < 0.05), unchanged in three and reduced in two tumour tissues. Aurora-C protein was increased by 9.2 +/- 2.90 fold (P < 0.05), suggesting that post-transcriptional mechanisms modulate its expression. In conclusion, we demonstrated that expression of Aurora kinases is deregulated in seminomas, suggesting that they may play a role in the progression of testicular cancers.

Expression of Fas and Fas ligand in human testicular germ cell tumours.

In the present study, we analysed the expression of Fas ligand (FasL) and its cognate receptor Fas in 14 seminomatous testicular germ cell tumours (TGCT) and six normal testicular tissues obtained following orchiectomy. Tissue samples have been processed to prepare either total RNA or protein extracts or fixed and embedded in paraffin for immunohistochemistry (IHC) experiments. Quantitative RT-PCR experiments demonstrated in TGCT a significant (p < 0.01) increase of the FasL mRNA expression of 21.1 +/- 5.4 fold, with respect to normal tissues. On the contrary, in the same cancer tissues, the levels of Fas mRNA were significantly (p < 0.01) reduced to 0.27 +/- 0.06 fold. These observations were confirmed in western blot experiments showing a significant increase of FasL and a concomitant decrease of Fas proteins in testicular cancer tissues, with respect to normal testis. Moreover, IHC experiments showed a strong FasL immuno-reactivity in six out of eight TGCT samples analysed, while Fas immuno-positivity was found in cancer cells of only two TGCT tissues. In addition, in all tumour samples, infiltrating lymphocytes were Fas positive. However, no correlation could be observed between Fas or FasL mRNA variations and clinical parameters such as patient's age, TNM stage or tumour size. We also compared the serum levels of soluble FasL (sFasL) of 15 patients affected by seminomatous TGCT, of four patients with non-seminomatous TGCT and six age-matched healthy males. No significant differences in sFasL serum level could be identified. In conclusion, our data demonstrated that the majority of seminomas are characterized by an increased expression of FasL and a concomitant reduction of Fas, with respect to human normal testis, and that sFasL serum level is not a tumour marker for patients affected by TGCT.

HPV genotypes concordance between sex partners.

The HPV genotype concordance in the sexual couples could support the sexual viral transmission of HPV infection. The present study contains a case-report of a stable Italian sex couple harbouring the same five HPV genotypes in their genital samples. The female partner, affected by vulvar condilomatosis, evidenced positivity in her cervicovaginal scraping with high risk HPV DNA Hybrid Capture 2 test and was negative at liquid-based performed Pap Test and at colposcopic examination. The male partner was clinically healthy regarding his external genitalia. In both male and female genital scrapings, the following HPV genotypes were detected by means of a PCR-based assay: 6, 16, 53, 73 and 84. This considerably high genotype concordance does not appear to be casual and supports, in our opinion, the hypothesis that genital HPV types are sexually transmitted agents

Status of the p53, p16, RB1, and HER-2 genes and chromosomes 3, 7, 9, and 17 in advanced bladder cancer: correlation with adjacent mucosa and pathological parameters.

AIMS: To evaluate a panel of well known genetic alterations for frequency of changes in bladder cancer that could be considered genomic instability determinants or adjunctive prognostic predictors. METHODS: Fluorescence in situ hybridisation analysis was performed to evaluate chromosomes 3, 7, 9, and 17 and the 9p21 (p16), 17p13.1 (p53), 13q14 (RB1), and 17q11.2 (HER-2) chromosomal loci in 48 muscle invasive bladder cancer specimens and the adjacent normal mucosa. RESULTS: There were significant differences between the frequency of chromosome 7 monosomy/polysomy and 17 monosomy in the two groups (tumours and adjacent mucosa) (p = 0.004, p = 0.037, and p = 0.015, respectively). There were no differences in the frequency of gene deletions between tumours and the adjacent mucosa. 17q11.2 amplification was found in 14.5% of tumours examined, but not in the non-malignant epithelium. Chromosome 3, 7, and 17 monosomy and the RB1 heterozygous deletion were significantly associated with stage T3-4 (p = 0.03, p = 0.04, p = 0.04, and p = 0.03, respectively). CONCLUSIONS: These results demonstrate the importance of chromosomes 3, 7, and 17 and gene alterations in bladder cancer progression, highlighting their usefulness as prognostic markers. Larger studies with longterm follow up of these patients are needed to determine the validity and clinical relevance of these genetic findings, and molecular prognostic markers should be incorporated into phase II and III trials to define their roles in predicting clinical outcome.

[Benign metastasizing pulmonary leiomyoma: description of a case and review of the literature]. / Leiomioma metastatizzante benigno nel polmone: descrizione di un caso e revisione della letteratura.

We report the case of a 48-year-old woman with multiple benign metastasizing leiomyomas in both lungs and a single retroperitoneal leiomyoma. The patient underwent surgical resection of the lesions 15 years after total hysterectomy for uterine myomas. Immunohistochemical studies performed on the retroperitoneal and pulmonary neoplasms showed them to be of mesenchymal derivation with smooth muscle differentiation; the samples were negative for HMB-45. Some histogenetic hypotheses for the multiple leiomyomas are presented.

Left atrial leiomyosarcoma: double occurrence and double recurrence--report of one case.

In October 1996 a 67-year-old man underwent transthoracic and transesophageal echocardiography (TEE) because of dyspnea on exertion and was found to have 2 left atrial cardiac masses. The 2 masses were surgically removed from the atrium and showed histopathologic and ultrastructural features of a leiomyosarcoma. Seven months later a double recurrence of left atrial masses was found with TEE; the patient refused surgery and decided instead to receive chemotherapy. In May 1998 he was in stable condition (New York Heart Association class III), but a further growth of the 2 left atrial masses was observed at TEE. We describe the echocardiographic features of the 2 cardiac masses and the clinical and prognostic implications.

Mucins in prostatic intra-epithelial neoplasia and prostatic carcinoma.

A histochemical study of 27 well-differentiated prostatic carcinoma cases associated with prostatic intra-epithelial neoplasia (PIN) was carried out. In the histological areas examined a decreasing production of neutral mucin was found as follows: normal prostatic tissue (70%), prostatic carcinoma (55%), PIN 1 (50%), PIN 2 (30%) and PIN 3 (15%). A progressively increasing content of acidic mucin (AB 2.5) was observed in the areas of PIN 3 (25%), PIN 2 (35%) and prostatic carcinoma (70%), while it was absent in the areas of PIN 1 and normal prostatic tissue. Acidic mucin sulphated type (AB 1) was secreted only in PIN 3 (5%) and prostatic carcinoma (10%) areas. These data were correlated with the proliferative activity of PIN. It is postulated that the lower neutral mucin production by PIN 3 can be linked to the higher proliferative activity of this lesion. Moreover, the acidic mucin secretion by PIN and prostatic carcinoma is considered to be a further feature of these lesions.

[Correlation between the nucleolar organizer region in adenocarcinoma of the prostate and the Gleason system]. / Correlazione tra le regioni organizzatrici del nucleolo nell'adenocarcinoma della prostata ed il sistema di Gleason.

Using a silver staining, technique, Nucleolar Organizer Region-associated proteins (NORs) were studied on paraffin sections of 25 resected prostatic adenocarcinomas classified with Gleason grading and 11 hyperplastic lesions. Then 7 inclusions was selected for each grade of Gleason system and 7 inclusions of normal prostatic tissue. The mean numbers of argyrophilic nucleolar organizer regions (AgNORs) increased significantly (P less than 0.01) from normal prostatic tissue to Gleason 5. The data indicate that AgNORs counts may help distinguish between each grade of Gleason system. It was concluded that the AgNOR technique provides a significant kinetic evaluation of prostatic adenocarcinoma and its prognostic study.

[Nucleolus organizer regions in prostatic intraepithelial neoplasm]. / Le regioni organizzatrici del nucleolo nella neoplasia intraepiteliale prostatica.

Using a silver staining technique, Nucleolar Organizer Region-associated proteins (NORs) were evaluated on paraffin sections of 16 resected prostatic adenocarcinomas stage A1. Then 30 histological areas was selected which comprised 6 areas for each grade of Prostatic Intraepithelial Neoplasia: PIN 1, PIN 2, PIN 3, 6 areas of normal glandular prostatic epithelium and 6 areas of well differentiated prostatic adenocarcinoma (Gleason I). The mean numbers of argyrophilic nucleolar organizer regions (AgNORs) increased from normal glandular prostatic epithelium to PIN 3, while the mean numbers of well differentiated prostatic adenocarcinoma was similar to PIN 1. A statistically significant difference (P less than 0.01) for AgNORs was found between normal glandular epithelium, PIN 1, PIN 2 and PIN 3 and between PIN 3 and well differentiated adenocarcinoma. It was concluded that AgNORs counts provide to significant kinetic evaluation of PIN and prostatic adenocarcinoma besides to supply a better definition of PIN.

[Intraepithelial prostatic neoplasia: a new dysplastic lesion of the prostate]. / La neoplasia intraepiteliale prostatica: una nuova lesione displastica della prostata.

Prostatic Intraepithelial Neoplasia were studied retrospectively in 455 cases of surgically resected prostates: 387 benign hypertrophies and 68 adenocarcinomas. The frequency of PIN was highest among prostatic adenocarcinomas well differentiated while the benign hypertrophies showed a lower frequency and a moderate increase with age. The prognostic importance of this lesion in prostatic biopsies is discussed on account of its strong predictive value.

Glial-like cells in sympathetic neural crest derivatives during human embryogenesis. Detection by S-100 immunohistochemistry.

The present study investigates at the light and electron microscopic levels the possible presence and distribution during human development of glial-like satellite cells in sympathetic neural crest derivatives by S-100 immunohistochemistry. From the earliest stages investigated, immunostained cells were detected inside sympathetic migrating masses and at their periphery, where they constituted a continuous layer isolating sympathetic elements from mesenchymal cells. The detection and peculiar distribution of these glial-like cells in developing sympathetic tissue could open new perspectives in the study of events linked to the migration and differentiation of some neural crest derivatives.

Protein absorption by tubular mesonephric and metanephric structures in the human embryo. An immunohistochemical study.

The presence of different serum proteins in the cells of the proximal tubule of both meso- and metanephric nephrons in human embryos (7th-12th week of intrauterine life) was investigated using immunohistochemistry. Endogenous lysozyme, alpha-1-antitrypsin and ferritin were detected in mesonephric proximal tubules and, starting from the 8th week, also in metanephric proximal tubules. Our observations provide information concerning the appearance and distribution of tubular protein reabsorption during the early stages of development.

Immunohistochemical distribution of S-100 antigen in the urinary system of man and rat.

The immunohistochemical distribution of S-100, a protein originally isolated from the brain, has been investigated at the light and electron microscopic levels in rat and man urinary systems. In both species the antigen essentially exhibited the same location, restricted, with different degrees of staining, to certain cells in the kidney, i.e. collecting tubules, thin limbs of Henle's loop and renal papillae.

A case of mucinous cystadenocarcinoma of the ovary associated with virilization: pre and post-operative steroid plasma levels.

A premenopausal woman with a mucinous carcinoma of one ovary, and a mucinous adenoma of the other, together with secondary virilization, is reported. Preoperative levels of androstenedione, testosterone and dehydroepiandrosterone sulphate were high, suggesting the presence of a virilizing tumor. Preoperative plasma estrone (E1), but not estradiol (E2), was elevated along with inversion of the E2/E1 ratio, suggesting a peripheral origin of the estrogens. FSH and LH plasma concentrations were low. After bilateral ovariectomy, levels of all steroids measured significantly decreased and gonadotropins rose to the postmenopausal range.