Diagnostic and Prognostic Value of ProEx C and GLUT1 in Melanocytic Lesions.

BACKGROUND: Melanoma is one of the most aggressive types of skin cancer. The purpose of this study was to evaluate the use of two biomarkers, ProEx C and glucose transporter isoform 1 (GLUT1), in the diagnosis and prognostication of melanoma. MATERIALS AND METHODS: We analyzed 129 melanomas and 59 benign nevi in a tissue microarray using immunohistochemical method with antibodies to topoisomerase IIα (TOP2A) and minichromosome maintenance complex component 2 (MCM2) using ProEx C and to GLUT1. RESULTS: The average proliferative index by ProEx C immunostain was significantly higher in melanomas (37.5%) compared to benign nevi (1.9%) as was the expression of GLUT1 (p<0.0001 respectively). Dermal mitosis was found to correlate positively with both ProEx C and GLUT1 (p=0.003 and p<0.001, respectively). Ulceration and tumor thickness positively correlated with GLUT1 expression (p=0.013 and p=0.033, respectively), but not with ProEx C staining. There was a significant association between increasing ProEx C index and stronger expression of GLUT1 (p<0.001). Kaplan-Meier disease-specific survival analyses indicated that patients whose melanoma exhibited expression of GLUT1 had a significantly lower rate of disease-specific survival than patients whose melanoma did not (p=0.039). However, staining by ProEx C did not show a prognostic significance in disease-specific survival. CONCLUSION: ProEx C and GLUT1 are potentially useful markers in differentiation of melanoma from nevi. Absence of GLUT1 expression in patients with primary melanoma predicts better survival.

Cell division cycle-associated protein 1 as a new melanoma-associated antigen.

Immune checkpoint inhibitors have increased the median survival of melanoma patients. To improve their effects, antigen-specific therapies utilizing melanoma-associated antigens should be developed. Cell division cycle-associated protein 1 (CDCA1), which has a specific function at the kinetochores for stabilizing microtubule attachment, is overexpressed in various cancers. CDCA1, which is a member of cancer-testis antigens, does not show detectable expression levels in normal tissues. Quantitative reverse transcription polymerase chain reaction and immunoblotting analyses revealed that CDCA1 was expressed in all of the tested melanoma cell lines, 74% of primary melanomas, 64% of metastatic melanomas and 25% of nevi. An immunohistochemical analysis and a Cox proportional hazards model showed that CDCA1 could be a prognostic marker in malignant melanoma (MM) patients. CDCA1-specific siRNA inhibited the cell proliferation of SKMEL2 and WM115 cells, but did not reduce the migration or invasion activity. These results suggest that CDCA1 may be a new therapeutic target of melanoma.

The metabolic microenvironment of melanomas: Prognostic value of MCT1 and MCT4.

BRAF mutations are known drivers of melanoma development and, recently, were also described as players in the Warburg effect, while this reprogramming of energy metabolism has been identified as a possible strategy for treating melanoma patients. Therefore, the aim of this work was to evaluate the expression and prognostic value of a panel of glycolytic metabolism-related proteins in a series of melanomas. The immunohistochemical expression of MCT1, MCT4, GLUT1, and CAIX was evaluated in 356 patients presenting melanoma and 20 patients presenting benign nevi. Samples included 20 benign nevi, 282 primary melanomas, 117 lymph node and 54 distant metastases samples. BRAF mutation was observed in 29/92 (31.5%) melanoma patients and 17/20 (85%) benign nevi samples. NRAS mutation was observed in 4/36 (11.1%) melanoma patients and 1/19 (5.3%) benign nevi samples. MCT4 and GLUT1 expression was significantly increased in metastatic samples, and MCT1, MCT4 and GLUT1 were significantly associated with poor prognostic variables. Importantly, MCT1 and MCT4 were associated with shorter overall survival. In conclusion, the present study brings new insights on metabolic aspects of melanoma, paving the way for the development of new-targeted therapies.

Prognostic Significance of Nuclear Phospho-ATM Expression in Melanoma.

UV radiation induced genomic instability is one of the leading causes for melanoma. Phosphorylation of Ataxia Telangiectasia Mutated (ATM) is one of the initial events that follow DNA damage. Phospho-ATM (p-ATM) plays a key role in the activation of DNA repair and several oncogenic pathways as well as in the maintenance of genomic integrity. The present study was therefore performed to understand the significance of p-ATM in melanoma progression and to correlate it with patient prognosis. Tissue microarray and immunohistochemical analysis were employed to study the expression of p-ATM in melanoma patients. A total of 366 melanoma patients (230 primary melanoma and 136 metastatic melanoma) were used for the study. Chi-square test, Kaplan-Meier, univariate and multivariate Cox regression analysis were used to elucidate the prognostic significance of p-ATM expression. Results revealed that both loss of, and gain in, p-ATM expression were associated with progression of melanoma from normal nevi to metastatic melanoma. Patients whose samples showed negative or strong p-ATM staining had significantly worse 5-year survival compared to patients who had weak to moderate expression. Loss of p-ATM expression was associated with relatively better 5-year survival, but the corresponding 10-year survival curve almost overlapped with that of strong p-ATM expression. p-ATM expression was found to be an independent prognostic factor for 5-year but not for 10-year patient survival. In conclusion our findings show that loss of p-ATM expression and gain-in p-ATM expression are indicators of worse melanoma patient survival.

Cellular localization of CIP2A determines its prognostic impact in superficial spreading and nodular melanoma.

Cancerous inhibitor of protein phosphatase 2A (CIP2A) is an important oncogene contributing to cancer progression partially by regulating cMYC and AKT. We examined CIP2A expression in cutaneous melanomas, its association with clinicopathological parameters and mapped molecular mechanisms regulated by CIP2A in vitro. CIP2A expression was analyzed by immunohistochemistry in 17 nevi, 132 primary melanomas and 49 metastases. Effects of siRNA-mediated down-regulation on proliferation, apoptosis and signaling pathways were assessed in melanoma cell lines. In superficial spreading melanomas (SSM), high nuclear CIP2A expression was associated with poor overall survival (OS) (P = 0.0018). Surprisingly, high cytoplasmic expression was related to improved relapse-free (P = 0.031) and OS (P = 0.014) in nodular melanomas (NM). In vitro experiments revealed that CIP2A can regulate proliferation and/or apoptosis partially through the PI3K/AKT pathway but also independently. In summary, CIP2A could represent a potential therapeutic target in SSM. However, in NM cytoplasmic CIP2A is associated with improved prognosis indicating that CIP2A has distinct, complex functions dependent on the molecular context and histological subtype. As seen in other cancer types, CIP2A can influence cMYC and AKT, but our data also suggest that in melanoma it has additional targets which need to be identified.

Outcome of patients with de novo versus nevus-associated melanoma.

BACKGROUND: Prior reports indicate a wide range of melanomas in histopathologic contiguity with a nevus, and an associated nevus has unclear prognostic implications in melanoma. OBJECTIVE: We sought to investigate the relationship among nevus-associated melanomas, sentinel lymph node status, and overall survival. METHODS: We conducted a retrospective analysis of 850 patients with cutaneous melanoma and sentinel lymph node removed at Massachusetts General Hospital from 1998 through 2008 and meta-analysis of the literature. RESULTS: Nevus-associated melanomas represented 28% (235/850) of cases and were significantly correlated with younger age (P = .03), truncal site (P = .0005), superficial spreading type (P < .0001), and absent ulceration (P = .005). There was no association with sentinel lymph node status (P = .94) and no survival difference between nevus-associated versus de novo melanoma (P = .41). Meta-analysis of over 4000 cases revealed a similar percentage of associated nevi (32%). LIMITATIONS: This was a retrospective study. CONCLUSIONS: Approximately 30% of melanomas are associated with a nevus. The presence of a nevus associated with a melanoma has no prognostic implication in sentinel lymph node status or overall survival.

Clinical characteristics and survival data of melanoma patients with nevus cell aggregates within sentinel lymph nodes.

Histopathologic differentiation of nevus cell aggregates and metastatic melanoma in lymph nodes is challenging. Patients with melanoma who had undergone sentinel lymph node (SLN) biopsy were evaluated using univariate and multivariate analyses as well as Kaplan-Meier statistics. Of the 651 patients, 50 (7.7%) had a nodal nevus in the SLN. In the logistic regression model, primary melanoma on the lower extremities proved to be the strongest independent negative predictor of nodal nevi with an odds ratio of 0.11 (95% confidence interval, 0.034-0.36; P = .0002). Overall 5-year survival (P = .17) and 5-year disease-free survival (P = .45) of patients with nodal nevi did not significantly differ from that of patients with negative SLNs. The frequency and anatomic localization of nodal nevi observed in the present study are in line with previous studies. Our 5-year survival data clearly demonstrate that nevus cell aggregates in lymph nodes have to be considered a benign condition even though it occurs in patients with melanoma. This study provides an indirect proof of validity and accuracy of current histopathologic methods for differentiation between nodal nevi and melanoma metastasis.

Disease progression and patient survival are significantly influenced by BRAF protein expression in primary melanoma.

BACKGROUND: Mutation of BRAF is a prevalent event in melanoma. Despite much attention to the role of BRAF mutation in melanoma, the status of BRAF protein expression and its significance in melanoma progression are unknown. OBJECTIVES: We investigated the BRAF expression level in different stages of melanocytic lesions and evaluated its correlation with clinicopathological features and patient survival. METHODS: Using tissue microarray, BRAF expression and its correlation with patient outcome was evaluated in 49 naevi samples and 370 patients with melanoma. We also evaluated the correlation of BRAF protein expression and V600E mutation using direct sequencing. RESULTS: Compared with naevi samples, BRAF expression was remarkably increased in primary melanomas and further increased in metastatic melanomas (P = 1·8 × 10(-11) ). High BRAF expression was significantly correlated with thicker tumours, ulceration and higher American Joint Committee on Cancer stages (P = 1·5 × 10(-7) , 1·5 × 10(-5) and 3·6 × 10(-13) , respectively). In cases of primary melanoma, patients with high BRAF expression had significantly worse overall (P = 0·009) and disease-specific 5-year survival (P = 0·007). While there was a trend for higher prevalence of BRAF V600E mutation in patients with high BRAF protein expression, no significant correlation was observed between protein expression and BRAF mutation. Furthermore, univariate Cox regression analysis confirmed high BRAF protein expression as a strong risk factor for poor patient survival in primary melanoma [hazard ratio (HR) 2·08 for overall survival; HR 2·39 for disease-specific survival]. CONCLUSIONS: Our data demonstrate that BRAF protein expression is significantly increased during melanoma progression. In addition, we revealed a novel prognostic value for BRAF protein expression in primary melanoma as it is significantly correlated with poor patient survival.

Kinesin family member 20A is a novel melanoma-associated antigen.

It has been shown recently that immunotherapy for advanced melanoma is effective. However, in order to improve the efficacy of immunotherapy, the identification of more specific melanoma-associated antigens is urgently needed. Kinesin family member 20A (KIF20A) has been reported to be a promising immunotherapeutic target for pancreatic cancer. To investigate the expression of KIF20A in melanoma, we performed quantitative reverse transcript (RT)-PCR and western blotting analyses of melanoma cell lines. We also investigated primary melanomas and naevus tissues with immunohistochemistry and real-time RT-PCR. KIF20A expression was detected in 59% of melanomas and 12% of naevi by immunohisto-chemistry, and 64% of melanomas and 60% of naevi by real-time RT-PCR. The primary melanomas that were positive for KIF20A showed a significantly greater thickness than those that were negative, and patients with KIF20A+ melanoma tended to develop recurrence earlier. These results suggest that immunotherapy with KIF20A may be a novel treatment option for advanced melanoma.

Cytoplasmic BRMS1 expression in malignant melanoma is associated with increased disease-free survival.

BACKGROUND/AIMS: Breast cancer metastasis suppressor 1 (BRMS1) blocks metastasis in melanoma xenografts; however, its usefulness as a biomarker in human melanomas has not been widely studied. The goal was to measure BRMS1 expression in benign nevi, primary and metastatic melanomas and evaluate its impact on disease progression and prognosis. METHODS: Paraffin-embedded tissue from 155 primary melanomas, 69 metastases and 15 nevi was examined for BRMS1 expression using immunohistochemistry. siRNA mediated BRMS1 down-regulation was used to study impact on invasion and migration in melanoma cell lines. RESULTS: A significantly higher percentage of nevi (87%), compared to primary melanomas (20%) and metastases (48%), expressed BRMS1 in the nucelus (p < 0.0001). Strong nuclear staining intensity was observed in 67% of nevi, and in 9% and 24% of the primary and metastatic melanomas, respectively (p < 0.0001). Comparable cytoplasmic expression was observed (nevi; 87%, primaries; 86%, metastases; 72%). However, a decline in cytoplasmic staining intensity was observed in metastases compared to nevi and primary tumors (26%, 47%, and 58%, respectively, p < 0.0001). Score index (percentage immunopositive celles multiplied with staining intensity) revealed that high cytoplasmic score index (≥ 4) was associated with thinner tumors (p = 0.04), lack of ulceration (p = 0.02) and increased disease-free survival (p = 0.036). When intensity and percentage BRMS1 positive cells were analyzed separately, intensity remained associated with tumor thickness (p = 0.024) and ulceration (p = 0.004) but was inversely associated with expression of proliferation markers (cyclin D3 (p = 0.008), cyclin A (p = 0.007), and p21Waf1/Cip1 (p = 0.009)). Cytoplasmic score index was inversely associated with nuclear p-Akt (p = 0.013) and positively associated with cytoplasmic p-ERK1/2 expression (p = 0.033). Nuclear BRMS1 expression in ≥ 10% of primary melanoma cells was associated with thicker tumors (p = 0.016) and decreased relapse-free period (p = 0.043). Nuclear BRMS1 was associated with expression of fatty acid binding protein 7 (FABP7; p = 0.011), a marker of invasion in melanomas. In line with this, repression of BRMS1 expression reduced the ability of melanoma cells to migrate and invade in vitro. CONCLUSION: Our data suggest that BRMS1 is localized in cytoplasm and nucleus of melanocytic cells and that cellular localization determines its in vivo effect. We hypothesize that cytoplasmic BRMS1 restricts melanoma progression while nuclear BRMS1 possibly promotes melanoma cell invasion.Please see related article: http://www.biomedcentral.com/1741-7015/10/19.

Mutations in GNA11 in uveal melanoma.

BACKGROUND: Uveal melanoma is the most common intraocular cancer. There are no effective therapies for metastatic disease. Mutations in GNAQ, the gene encoding an alpha subunit of heterotrimeric G proteins, are found in 40% of uveal melanomas. METHODS: We sequenced exon 5 of GNAQ and GNA11, a paralogue of GNAQ, in 713 melanocytic neoplasms of different types (186 uveal melanomas, 139 blue nevi, 106 other nevi, and 282 other melanomas). We sequenced exon 4 of GNAQ and GNA11 in 453 of these samples and in all coding exons of GNAQ and GNA11 in 97 uveal melanomas and 45 blue nevi. RESULTS: We found somatic mutations in exon 5 (affecting Q209) and in exon 4 (affecting R183) in both GNA11 and GNAQ, in a mutually exclusive pattern. Mutations affecting Q209 in GNA11 were present in 7% of blue nevi, 32% of primary uveal melanomas, and 57% of uveal melanoma metastases. In contrast, we observed Q209 mutations in GNAQ in 55% of blue nevi, 45% of uveal melanomas, and 22% of uveal melanoma metastases. Mutations affecting R183 in either GNAQ or GNA11 were less prevalent (2% of blue nevi and 6% of uveal melanomas) than the Q209 mutations. Mutations in GNA11 induced spontaneously metastasizing tumors in a mouse model and activated the mitogen-activated protein kinase pathway. CONCLUSIONS: Of the uveal melanomas we analyzed, 83% had somatic mutations in GNAQ or GNA11. Constitutive activation of the pathway involving these two genes appears to be a major contributor to the development of uveal melanoma. (Funded by the National Institutes of Health and others.).

Myeloid leukemia-1 expression in benign and malignant melanocytic lesions.

Myeloid leukemia-1 (Mcl-1) is an anti-apoptotic protein implicated in tumor progression. Its expression was found to be elevated in many types of human cancers and is correlated with tumor progression. The expression of Mcl-1 in melanoma is not fully understood. We investigated the expression of Mcl-1 in normal nevi, dysplastic nevi, primary melanoma and melanoma metastases by tissue microarray and immunohistochemistry. We found that Mcl-1 expression was significantly increased in dysplastic nevi, primary melanoma and melanoma metastases when compared to normal nevi, though the expression of Mcl-1 was decreased in metastatic melanoma when compared to dysplastic nevi. We did not find any correlation between Mcl-1 expression and melanoma patient survival. Our data suggest that Mcl-1 may play a critical role in the initiation of melanoma development.

Enhancement of lesion classification using divergence, curl and curvature of skin pattern.

BACKGROUND/PURPOSE: The observation that skin pattern tends to be disrupted by malignant but not by benign skin lesions suggests that measurements of skin pattern disruption on simply captured white light optical clinical (WLC) skin images could be a useful contribution to a diagnostic feature set. Previous work which generated a flow field of skin pattern using a measurement of local line direction and variation determined by the minimum eigenvalue and its corresponding eigenvector of the local tensor matrix to measure skin pattern disruption was computationally low cost and encouraging. This paper explores the possibility of extracting new features from the first and second differentiations of this flow field to enhance classification performance. METHODS: The skin pattern was extracted from WLC skin images by high-pass filtering. The skin line direction was estimated using a local image gradient matrix to produce a flow field of skin pattern. Divergence, curl, mean and Gaussian curvatures of this flow field were computed from the first and second differentiations of this flow field. The difference of these measures combined with skin line direction across the lesion image boundary was used as a lesion classifier. RESULTS: A set of images of malignant melanoma and benign naevi were analysed as above and the scatter plot in a two-dimensional dominant feature space using principal component analysis showed excellent separation of benign and malignant lesions. A receiver operating characteristic plot enclosed an area of 0.96. CONCLUSIONS: The experimental results show that the divergence, curl, mean and Gaussian curvatures of the flow field can increase lesion classifier accuracy. Combined with skin line direction they are promising features for distinguishing malignant melanoma from benign lesions and the methods used are computationally efficient which is important if their use is to be considered in clinical practice.

[The value of radiotherapy in treatment of meningeal melanocytoma]. / Stellenwert der Strahlentherapie bei der Behandlung des meningealen Melanozytoms.

BACKGROUND: Meningeal melanocytoma is described as rare benign lesion with a high risk of recurrence. There are no well-substantiated treatment recommendations in the literature. Only case reports have been published by now. PATIENTS AND METHODS: In 1997 a patient was irradiated for a recurrent spinal meningeal melanocytoma and 2 years later for brain metastases indicating malignant transformation. This case gave rise to a literature review for therapeutic options. All sufficiently documented cases published since 1972, when the term meningeal melanocytoma was established, were evaluated. Based on published and on original data recurrence and overall survival rates up to 5 years were calculated for three different therapeutic approaches, namely complete tumor resection, incomplete resection with subsequent radiotherapy, and incomplete resection alone. Statistical evaluation was performed using the chi 2 test and Kaplan-Meier-analysis. RESULTS: 53 patients (including our patient) met selection criteria. Complete tumor resection was superior to incomplete resection alone with lower recurrence (4-38% versus 50-92%) and better overall survival rates (86-95% versus 30-58%). After incomplete resection radiotherapy seemed to improve prognosis (recurrence 15-45%; overall survival 91-92%). Between complete resection and incomplete resection plus radiotherapy no significant differences were observed. CONCLUSIONS: For meningeal melanocytoma complete resection must be regarded as the best of the modalities compared. After incomplete resection radiotherapy should be considered, although a specific radiotherapeutic regimen cannot be recommended at present. However, for multiple cranial or spinal lesions total cranial irradiation or craniospinal irradiation is indicated.

Protein expression of the cell-cycle inhibitor p27Kip1 in malignant melanoma: inverse correlation with disease-free survival.

In the present study we analyzed, by immunohistochemistry, a panel of human melanomas for protein expression of the cyclin-dependent kinase (cdk) inhibitor p27Kip1 and evaluated whether deregulated expression correlates with clinical outcome for this type of cancer. We found that p27Kip1 was strongly expressed by normal melanocytes and benign nevi, whereas in malignant melanoma, a heterogeneous expression pattern was observed. In the case of nodular melanomas, the level of p27Kip1 was found to correlate significantly with the thickness of the tumor, with less protein expressed in thicker lesions. We also found that patients having tumors with fewer than 5% p27Kip1-staining cells had a significantly higher risk of early relapse of their disease compared with those expressing moderate or high levels. In contrast, the level of p27Kip1 did not correlate with tumor thickness or disease-free survival in patients with superficial spreading melanomas, suggesting that p27Kip1 may play different roles in these two major pathological subgroups of malignant melanoma. Furthermore, p27Kip1 did not appear to have an influence on overall survival for either subgroup. When we examined the combined effect of p21WAF1/CIP1 (another cdk inhibitor) and p27Kip1 on clinical outcome, we found that analysis of these two cdk inhibitors together may have greater prognostic potential than either alone. In conclusion, our results suggest that virtually complete loss of p27Kip1 protein expression has potential importance as a prognostic indicator of early relapse in patients with nodular melanoma The results, furthermore, underscore the value of analyzing multiple cell cycle regulatory proteins to obtain the most reliable indication of prognosis.

Antigen Ki-67 and c-myc oncogene as related to histoclinical parameters in pigmented skin lesions.

121 cases of cutaneous melanomas, 82 lymph nodes with melanoma metastases and 62 naevi were stained immunohistochemically for presence of Ki-67 antigen and c-myc oncogene. A significant correlation between expression of investigated markers, and survival time as well as presence of metastases, in cutaneous melanomas was found (p < 0.05). The role of Ki-67 antigen for differential diagnosis between naevi and melanomas is also demonstrated.

Significance of P-53 antigen in malignant melanomas and naevi of the head and neck area.

DO-7 antibody against p-53 antigen was applied for investigation of melanomas of facial skin (25 cases), oral cavity (17 cases), eye (18 cases) and naevi (25 cases). The p-53 index value (% of p-53 positive cells) was correlated with the thickness of the tumour, the presence of metastases and survival time. The difference in p-53 index between naevi and melanomas was statistically significant (p < < 0.01). A significant correlation was found between the p-53 index value and the thickness of the tumour, the presence of metastases and follow-up for patients with skin, oral and ocular melanomas. The possible diagnostic and prognostic significance of p-53 antigen in melanomas and naevi of the head and neck area is discussed.

The risk of melanoma in patients with congenital nevi: a cohort study.

BACKGROUND: Giant congenital nevi are associated with a greatly increased risk of melanoma, but this has not been quantified. Smaller congenital nevi are believed by some authors to be associated with increased risk, but this is uncertain and needs to be clarified. OBJECTIVE: Our purpose was to analyze cause-specific mortality and cancer incidence risks in patients with congenital nevi according to size of the nevi. METHODS: We followed up 265 patients with congenital nevi first treated at the Hospital for Sick Children or at St. John's Hospital in London during 1950 to 1984 for mortality to mid-1993 and for cancer incidence from 1971 to 1989. Mortality and cancer incidence rates in the cohort were compared with expectations from national mortality and cancer incidence rates by sex, age, and calendar period. RESULTS: Among the 33 patients with a congenital nevus covering at least 5% of the body area, two melanomas occurred during follow-up; both were fatal. The relative risk of melanoma mortality in these patients was 1046 (95% confidence interval, 127 to 3779). In the remaining 232 patients, 68 of whom had a nevus covering 1% to 4% of the body, and 164 with nevi smaller than 1% of body area, no melanomas occurred (0.18 melanoma deaths expected). The difference in melanoma mortality risk between the group with a nevus covering at least 5% of the body and the group with smaller nevi was significant (p < 0.05). There was not a significantly increased risk of nonmelanoma mortality or of nonmelanoma cancer incidence overall in the cohort, although two lymphohematopoietic malignancies occurred. CONCLUSION: The data show the large risk of melanoma in patients with nevi covering more than 5% of the body surface area. The results do not support the hypothesis of greatly increased risk in persons with congenital nevi smaller than this, but because the confidence intervals of the result were wide, the data are compatible with a sizable risk. Much larger studies than those that have so far been undertaken, or combined analysis of data from several studies, are needed to quantify more precisely the risk of melanoma in relation to size of nevi and to determine the appropriate clinical management of these lesions.

Role of the argyrophilic nucleolar organizer regions in tumor detection and prognosis.

The expression of the argyrophilic nucleolar organizer regions (AgNORs) has been analyzed in renal, bladder, and pharyngeal carcinomas, multiple myeloma (MM), and skin melanocytic lesions to clarify their role in tumor detection and prognosis. Sections from formalin-fixed, paraffin-embedded biopsies were stained with the method of Ploton; the mean AgNOR number per nucleus (AgNOR count) and their distribution (configuration) were assessed examining 100 neoplastic cells. AgNOR counts and histologic grade were highly associated in bladder urotheliomas (6.01 for grade 1 [G1], 7.69 for G2, 13.35 for G3; p < 0.00001) and MM (3.18 for G1, 4.36 for G2, 6.13 for G3; p < 0.0001); they were not associated in renal cell carcinomas (5.35 for G1, 5.92 for G2, 7.99 for G3; p = 0.132) and pharyngeal carcinomas (11.1 for G2, 10.27 for G3; p = 0.08). AgNOR number was also related to the degree of malignancy in melanocytic lesions (2.93 for common blue nevus, 2.89 for benign nevus [BN], 3.69 for cellular blue nevus [CBN], 7.71 for malignant melanoma, and 8.33 for malignant cellular blue nevus [MCBN]; p < 0.00001). Association between AgNOR counts and pathologic stage was found in bladder carcinomas (6.43 for pTa, 10.19 for pT1, 12.57 for pT2-4; p < 0.00001) and MM (3.06 for cases with percentage of bone marrow plasma cells [BMPC%] < or = 20, 4.28 for BMPC% 21 to 50, 5.14 for BMPC% > 50; p < 0.0001]; no correlation was found in pharyngeal (11.18 for T1, 10.08 for T2, 10.68 for T3, 11.47 for T4; p = 0.18) or renal cell carcinomas (6.06 for pT2, 6.31 for pT3; p = 0.78). Few, large and grouped AgNORs were found in well-differentiated bladder carcinomas, MM, and benign melanocytic lesions; numerous, small and dispersed AgNORs were seen in poorly differentiated bladder, renal and pharyngeal carcinomas, MM and malignant melanocytic lesions. Significant association with prognosis was found in pharyngeal carcinomas (5-year survival: 68% for cases with < or = 10.31 AgNOR/cell, 20% for cases with > 10.31 AgNORs) and MM (5-year survival: 46% for cases with < or = 4.62 AgNOR/cell, 7% for cases with > 4.62 AgNORs; in MM the configuration too was related to prognosis: median of survival 72 months for tightly grouped, 16 for partially grouped, and 11 for dispersed AgNORs). Our results indicate that AgNOR number and configuration are useful in detection and prognosis of some neoplasias. They permit a rapid evaluation of morphology and tumor cell kinetics even on small biopsies.(ABSTRACT TRUNCATED AT 400 WORDS)