Assessment of HER-2 status in invasive breast cancer in Brazil.

OBJECTIVE: To characterize the frequency of HER-2-positive breast cancer in Brazil. METHOD: In this prospective observational study, we first ascertained the HER-2 status of invasive breast cancer specimens by automated immunohistochemistry (IHC). For specimens classified as 2+ by IHC, we performed in situ hybridization (ISH). RESULTS: From February, 2011 to December, 2012, 1,495 breast specimens were registered, and 1,310 samples collected at 24 centers were analyzed. Median patient age was 54 years, and the majority of samples were obtained from segmental (46.9%) or radical mastectomy (34.4%). The predominant histological type was invasive breast carcinoma of no special type (85%), 64.3% had tubule formation (grade 3), and estrogen/progesterone receptors (ER/PR) were positive in 77.4/67.8% of the specimens analyzed, respectively. Using IHC, we found a negative HER-2 status (0 or 1+) in 72.2% of specimens, and 3+ in 18.5%; the 9.3% scored as 2+ were further analyzed by ISH, of which 15.7% were positive (thus, 20.0% of samples were HER-2- -positive by either method). We found no association between HER-2 scores and menopausal status or histological type. Tumors classified as 3+ came from younger patients, and had higher histological grade and less frequent expression of ER/PR. In the North region of Brazil, 34.7% of samples were 3+, with lower frequencies in the other four regions of the country. CONCLUSION: Our findings provide estimates for the frequency of HER-2 positivity in Brazil and raise the hypothesis that biological differences may underlie the different distribution of breast-cancer phenotypes among different Brazilian regions.

Assessment of HER-2 status in invasive breast cancer in Brazil / Avaliação de HER-2 no câncer de mama invasivo no Brasil

Summary Objective: To characterize the frequency of HER-2-positive breast cancer in Brazil. Method: In this prospective observational study, we first ascertained the HER-2 status of invasive breast cancer specimens by automated immunohistochemistry (IHC). For specimens classified as 2+ by IHC, we performed in situ hybridization (ISH). Results: From February, 2011 to December, 2012, 1,495 breast specimens were registered, and 1,310 samples collected at 24 centers were analyzed. Median patient age was 54 years, and the majority of samples were obtained from segmental (46.9%) or radical mastectomy (34.4%). The predominant histological type was invasive breast carcinoma of no special type (85%), 64.3% had tubule formation (grade 3), and estrogen/progesterone receptors (ER/PR) were positive in 77.4/67.8% of the specimens analyzed, respectively. Using IHC, we found a negative HER-2 status (0 or 1+) in 72.2% of specimens, and 3+ in 18.5%; the 9.3% scored as 2+ were further analyzed by ISH, of which 15.7% were positive (thus, 20.0% of samples were HER-2- -positive by either method). We found no association between HER-2 scores and menopausal status or histological type. Tumors classified as 3+ came from younger patients, and had higher histological grade and less frequent expression of ER/PR. In the North region of Brazil, 34.7% of samples were 3+, with lower frequencies in the other four regions of the country. Conclusion: Our findings provide estimates for the frequency of HER-2 positivity in Brazil and raise the hypothesis that biological differences may underlie the different distribution of breast-cancer phenotypes among different Brazilian regions.

Resumo Objetivo: Estimar a frequência de câncer de mama positivo para HER-2 no Brasil. Método: Neste estudo observacional e prospectivo, verificamos o escore de HER-2 de espécimes de câncer de mama invasivo por imuno-histoquímica automatizada (IHQ). Para amostras classificadas como 2+ por IHQ, fizemos hibridização in situ (HIS). Resultados: De fevereiro de 2011 a dezembro de 2012, 1.495 espécimes de mama foram registrados, e 1.310 amostras coletadas por 24 centros foram analisadas. A idade mediana das pacientes foi 54 anos, e a maioria das amostras foram obtidas a partir de mastectomia segmentar (46,9%) ou radical (34,4%). O tipo histológico predominante foi o carcinoma invasivo da mama, sem tipo especial (85%); 64,3% tinham formação de túbulos (grau 3); e os receptores de estrógeno (RE)/progesterona (RP) foram positivos em 77,4%/67,8% das amostras analisadas. Por IHQ, encontramos HER-2 negativo (0 ou 1+) em 72,2% das amostras, e 3+ em 18,5%; os 9,3% de casos classificados como 2+ foram analisados por HIS, e 15,7% deles foram positivos (assim, 20,0% das amostras foram positivas para HER-2 por qualquer método). Não encontramos associação entre escores de HER-2 e estado menopausal ou tipo histológico. Tumores classificados como 3+ vieram de pacientes mais jovens, tinham maior grau histológico e foi menos frequente a expressão de RE/RP. Na região Norte do Brasil, 34,7% das amostras foram 3+, com frequências mais baixas nas outras quatro regiões do país. Conclusão: Nossos resultados permitem estimar a frequência de positividade do HER-2 no Brasil, gerando a hipótese de que pode haver diferenças biológicas subjacentes à distribuição dos fenótipos de câncer de mama entre as diferentes regiões brasileiras.

Raf Kinase Inhibitor Protein Expression and Prognostic Value in Soft Tissue Sarcomas.

OBJECTIVE: Soft tissue sarcomas (STSs) are heterogeneous tumors displaying multiple and complex molecular abnormalities with no specific pattern. Despite current therapeutic advances, the patients with STS still have a poor outcome, which makes it necessary to find out new prognostic markers. The Raf kinase inhibitory protein (RKIP) has been associated with prognosis in several human neoplasms; however, its role in STS is unknown. METHODS: In the present study RKIP expression was assessed by immunohistochemistry in a series of 87 STSs, and its expression profile was associated with the patients' pathological parameters. RESULTS: We found that RKIP is expressed in the cytoplasm of the great majority of cases, and absent in only approximately 18% of cases (16/87). Importantly, we observed that loss of RKIP expression was associated with poor outcome, constituting an independent prognostic marker. CONCLUSION: This is the first study assessing RKIP expression levels in STS. We showed that loss of RKIP expression is present in a small subset of cases; however, its absence was associated with poor survival and may be a potential marker for STS prognosis.

Retrospective analysis of breast cancer prognosis among young and older women in a Brazilian cohort of 738 patients, 1985-2002.

Invasive breast cancer (BC) is infrequent among women aged ≤40 years, however, the disease outlook in these younger patients is generally worse than among older women. The present study aimed to compare socio-demographic, clinical and pathological characteristics, and their association with long-term survival, between two random cohorts of young (≤40 years) and older (50-69 years) Brazilian patients with BC. The cohort comprised of 738 randomly selected women who were diagnosed with BC at Barretos Cancer Hospital, Pio XII Foundation (Barretos, Brazil) between January 1985 and December 2002; the patients included young women (n=376) and older women (n=362). The current analysis suggested that BC in young women is associated with numerous pathological features of aggressiveness. Second cancer and bilateral BC were independent predictors of a poor outcome in the younger group. Furthermore, C-erB-2 was positively correlated with poor outcome in the older group, whereas estrogen receptor status and TNM stage were associated with disease prognosis in both groups. The overall survival rates of the two age groups were similar except when analyzed according the treatment period (1997-2002). Although patients aged ≤40 years harbored tumors with more aggressive clinicopathological characteristics, these characteristics were not independent predictors of overall survival. The present study indicates that medical advances associated with prevention of breast cancer may improve screening programs, which may therefore increase early diagnosis and subsequently lower mortality rates.

Molecular profiling, including TERT promoter mutations, of acral lentiginous melanomas.

Acral lentiginous melanoma (ALM) is the less common subtype with singular characterization. TERT (human telomerase reverse transcriptase) promoter mutations have being described as recurrent in melanomas and infrequent in ALM, but their real incidence and clinical relevance is unclear. The objectives of this study were to describe the prevalence of TERT promoter mutations in ALM, and correlate with the molecular profile of other drive genes and clinical features. Sixty-one samples from 48 patients with ALM were analyzed. After DNA isolation, the mutation profiles of the hotspot region of BRAF, NRAS, KIT, PDGFRA, and TERT genes were determined by PCR amplification followed by direct Sanger sequencing. KIT, PDGFRA, and VEGFR2 gene amplification was performed by quantitative PCR. Clinical information such as survival, clinical stage, and Breslow tumor classification were obtained from medical records. TERT promoter mutations were found in 9.3% of the cases, BRAF in 10.3%, NRAS in 7.5%, KIT in 20.7%, and PDGFRA in 14.8% of ALM. None of the cases showed KIT, PDGFRA, or VEGFR2 gene amplification. We found an association between KIT mutations and advanced Clark level (IV and V, P=0.043) and TERT promoter mutations with low mitotic index. No other significant associations were observed between mutation profile and patients' clinical features nor survival rates. Oncogenic TERT promoter mutations are present in a fraction of ALMs. No relevant associations were found between TERT mutation status and clinical/molecular features nor survival. Mutations of KIT and PDGFRA are the most common genetic alterations, and they can be therapeutic targets for these patients.

A microRNA signature profile in EBV+ diffuse large B-cell lymphoma of the elderly.

Currently, there is no characteristic microRNA (miRNA) expression pattern in Epstein-Barr virus+ diffuse large B-cell lymphoma of the elderly (EBV+DLBCLe). This study aims to characterize a signature profile and identify miRNAs that can be used as biomarkers and alternative therapeutic targets for EBV+DLBCLe. Seventy-one DLBCL patients aged 50 years and older were included and four EBV+ and four EBV- samples were analyzed in two miRNA array platforms (pilot study). A larger multicenter cohort (29 EBV+DLBCLe and 65 EBV-DLBCL patients) was used to validate the results by real-time polymerase chain reaction. In the pilot study, 9% of DLBCL were EBV+DLBCLe by in situ hybridization. In multicenter study, EBV+DLBCLe group showed a predominance of non-germinal center B-cell origin. Overall survival duration of EBV+DLBCLe was significantly inferior to that of EBV-DLBCL patients. We found 10 deregulated miRNAs in the two groups, but only seven were statistically different. We confirmed overexpression of hsa-miR-126, hsa-miR-146a, hsa-miR-146b, hsa-miR-150, and hsa-miR-222 and underexpression of hsa-miR-151 in EBV+DLBCLe cases compared to EBV-DLBCL cases. Hsa-miR-146b and hsa-miR-222 showed high specificity for identifying EBV+DLBCLe. The present study proposed a miRNA signature for EBV+DLBCLe and our findings suggest that hsa-miR-146b and hsa-miR-222 could be biomarkers and therapeutic targets.

Automated screening of conventional gynecological cytology smears: feasible and reliable.

OBJECTIVES: We tested the ability of automated screening in processing conventional gynecological cytology smears and its efficacy in assessing sample adequacy and stratifying cases for risk of malignancy. STUDY DESIGN: Cases were retrospectively selected, including unsatisfactory samples and slides with various sorts of artifacts. Automated screening was performed using the FocalPoint GS Imaging System (Becton Dickinson, Franklin Lakes, N.J., USA), with classification into five quintiles. For agreement purposes, cases were grouped into high risk for malignancy (quintiles 1 and 2) and low risk for malignancy (quintiles 3, 4 and 5). RESULTS: A total of 120 cases (median age 37.5 years, range 18-85) were included in the study. Eighty-three cases (69.2%) could be successfully classified into quintiles. When divided by risk, 31 cases were placed in the high-risk and 52 in the low-risk group. The overall sensitivity and specificity of the automated analysis was 100 and 70.3%, respectively. CONCLUSIONS: Automated analysis could analyze the majority of conventional smears, including one case previously screened as unsatisfactory. All malignant and high-grade lesions were correctly classified into the high-risk group. Broad use of this automation system could potentially decrease screening time and augment the efficacy in detecting precursor neoplastic changes in cervical cytology smears.

The significance of augmented high-grade squamous intraepithelial lesion detection on pap test examination: partial results from the RODEO study team.

OBJECTIVE: This study sought to ascertain the significance of augmented high-grade squamous intraepithelial lesion (HSIL) detection by Pap test using both conventional smear and liquid-based cytology (LBC) in a high-risk population. STUDY DESIGN: We conducted a direct-to-vial study to compare the diagnostic performance of Pap smear versus LBC in a high-risk population of women referred for colposcopy at a gynecologic ambulatory clinic at the Barretos Cancer Hospital in Brazil during 2011. RESULTS: The detection of both low-grade squamous intraepithelial lesions (LSILs) and HSILs was significantly greater (p = 0.04 and p = 0.033, respectively) in the LBC arm [84 LSIL cases (5.7%) and 148 HSIL cases (10.1%)] than in the conventional smear arm [66 LSIL cases (4.1%) and 126 HSIL cases (7.9%)]; however, no differences were found for invasive squamous carcinoma or adenocarcinoma (p = 0.678). Of 3,071 women who were examined cytologically (1,604 conventional preparations and 1,467 LBC) and colposcopically, biopsies were available for 279 conventional preparations (17.6%) and 325 LBC preparations (22.2%). No significant differences were found between the methods with respect to diagnostic performance. CONCLUSION: LBC was significantly superior to conventional smears for the detection of LSILs and HSILs, but these results did not influence biopsy confirmation. Both methods showed similar performances with high positive predictive values but very low sensitivities.

Performance and reproducibility of gynecologic cytology interpretation using the FocalPoint system: results of the RODEO Study Team.

OBJECTIVES: To assess whether automated screening in the cytologic examination of Papanicolaou smear slides results in smaller margins of error than manual screening. METHODS: We compared cytotechnologists' performance and reproducibility of manual and automated screening of 10,165 consecutive cervical cytology slides examined at Barretos Cancer Hospital using the FocalPoint system. RESULTS: In total, 83% of atypical squamous cells of undetermined significance and greater were classified as quintiles 1 and 2; no high-grade squamous intraepithelial lesions and greater were observed in quintile 5. No statistically significant differences were found between manual and automated screening, using cervical biopsy specimens as the gold standard. CONCLUSIONS: FocalPoint safely screened high-grade lesions, which can be valuable for high-workload routines.

Immunohistochemical analysis of mechanoreceptors in the human posterior cruciate ligament: association with aging male.

INTRODUCTION: The aim of this study was to evaluate the association between frequency of the posterior cruciate ligament (PCL) mechanoreceptors and age in men. METHODS: Nineteen normal right knees harvested from human male cadavers were evaluated. Age ranged from 17 to 64 years with a mean of 35 years old. PCL was separated for sampling in femoral and tibial portions. Topographic distribution and frequency within the ligament texture were determined employing the Pro-Image digital analysis system. Mechanoreceptors were counted and classified according to the criteria proposed by Freeman & Wyke. RESULTS: A total of 1820 mechanoreceptors were found, type II being the most frequent one. Analysis of the femoral portion of the ligament showed an equivalent predominance of Types II and IV mechanoreceptors. Tibial portion had a predominance of type II mechanoreceptors, followed by type IV. At this portion, receptors Types I and III were less commonly identified. CONCLUSION: In the tibial portion of the PCL, there is predominance of Type II mechanoreceptors followed by types IV, I and III mechanoreceptors, respectively. No relationship was found between the total number of mechanoreceptors and age in the femoral and tibial portions of the PCL.

Could alarmingly high rates of negative diagnoses in remote rural areas be minimized with liquid-based cytology? Preliminary results from the RODEO Study Team.

OBJECTIVE: It was the aim of this study to compare diagnostic performances of the BD SurePath™ liquid-based Papanicolaou test (LBC) and the conventional Papanicolaou test (CPT) in cervical samples of women from remote rural areas of Brazil. STUDY DESIGN: Specimens were collected by mobile units provided by Barretos Cancer Hospital. This report evaluates the manual screening arm of the RODEO study. Of 12,048 women seen between May and December 2010, 6,001 were examined using LBC and 6,047 using CPT. RESULTS: Comparative (LBC vs. CPT) outcomes were: all abnormal tests, 2.1 versus 1.0%; ASC-US (atypical squamous cells of unknown significance), 0.7 versus 0.1%; ASC-H (atypical squamous cells with possible high-grade squamous intraepithelial lesions) and AGC (atypical glandular cells), 0.4 versus 0.3%; LSIL (low-grade squamous intraepithelial lesions), 0.7 versus 0.3%; HSIL (high-grade squamous intraepithelial lesions), 0.4 versus 0.2%, and unsatisfactory, 0.03 versus 0.08%. The LBC arm detected significantly more lesions (ASC-US+) than CPT (p < 0.001); however, when we divided the diagnoses into two groups, ASC-H- (negative/ASC-US/LSIL) and ASC-H+ (ASC-H/AGC/HSIL), the difference was not statistically important (p = 0.213). CONCLUSIONS: With inherent difficulties in patient recruitment and patient compliance with cancer screening, best test performance including human papillomavirus test capability are vitally necessary in Brazil's struggle to reduce cervical cancer.

Co-expression of monocarboxylate transporter 1 (MCT1) and its chaperone (CD147) is associated with low survival in patients with gastrointestinal stromal tumors (GISTs).

Monocarboxylate transporters (MCTs) have been described to play an important role in cancer, but to date there are no reports on the significance of MCT expression in gastrointestinal stromal tumors (GISTs). The aim of the present work was to assess the value of MCT expression, as well as co-expression with the MCT chaperone CD147 in GISTs and evaluate their clinical-pathological significance. We analyzed the immunohistochemical expression of MCT1, MCT2, MCT4 and CD147 in a series of 64 GISTs molecularly characterized for KIT, PDGFRA and BRAF mutations. MCT1, MCT2 and MCT4 were highly expressed in GISTs. CD147 expression was associated with mutated KIT (p = 0.039), as well as a progressive increase in Fletcher's Risk of Malignancy (p = 0.020). Importantly, co-expression of MCT1 with CD147 was associated with low patient's overall survival (p = 0.037). These findings suggest that co-expression of MCT1 with its chaperone CD147 is involved in GISTs aggressiveness, pointing to a contribution of cancer cell metabolic adaptations in GIST development and/or progression.

Lymphangiogenic VEGF-C and VEGFR-3 expression in genetically characterised gastrointestinal stromal tumours.

This study aimed to assess the distribution of VEGF-C and VEGFR-3 expression in gastrointestinal stromal tumours (GISTs), and to analyse the value of lymphatic vessel density (LVD) in a tumour that is believed to preferentially metastasize through blood vessel conduits. A panel of immunohistochemical antibodies was used to evaluate 51 cases of genetically characterised GISTs: VEGF-C, VEGFR-3, D2-40 (for LVD assessment) and CD31 (for blood vessel density--BDV--assessment). The results were correlated with the clinical-pathological data. The large majority of cases (86.2%; 44/51) showed a mutation of the KIT gene, most of them (72.5%; 37/51) revealing mutations in exon 11. VEGFR-3 was predominantly expressed in KIT mutated GISTs (p=0.019). High LVD was correlated with the absence of metastasis (p=0.010) and high BVD showed a positive correlation with the occurrence of metastasis (p=0.049). The strong expression of VEGF-C and VEGFR-3 in GIST's cells was not correlated with the clinical parameters of aggressiveness, nor with high LVD.

Prognostic and risk factors in patients with locally advanced cutaneous squamous cell carcinoma of the trunk and extremities.

55 patients with advanced cutaneous squamous cell carcinoma (CSCC) of the trunk and extremities were studied. A Tissue Microarray was constructed using immunohistochemistry to quantify expression of the HER family, E-cadherins, and podoplanin. Clinical and histopathological factors related to lymph node metastasis and prognosis were also established. Primary tumor positivity was 25.5% for EGFR, 87.3% for HER-3, and 48.1% for HER-4. Metastases were positive for EGFR in 41.7%, for HER-3 in 83.3%, and HER-4 in 43.5%. HER-2 was negative in all samples. Membrane E-cadherin and cytoplasmic E-cadherin were positive in 47.3% and 30.2% of primary tumors and 45.5% and 27.3% of metastases, respectively. Podoplanin was positive in 41.8% of primary tumors and 41.7% of metastases. Intratumoral lymphocytic infiltrate was associated with lymph node metastasis. Patients with T3 tumors had better cancer-specific survival (CSS) than those with T4 tumors; patients with no lymph node involvement had better CSS than patients with N1 tumors. Undifferentiated tumors and hyperexpression of podoplanin were negative prognostic indicators on multivariate analysis.

Hiperparatireoidismo secundário: fatores prognósticos de recidiva atribuída ao implante após paratireoidectomia total e auto-implante / Secondary hyperparathyroidism: prognostic factors of graft-dependent recurrence after total parathyroidectomy adn parathyroid autotransplantation

Introdução: Quando o tratamento clínico do hiperparatireoidismo secundário falha, é recomendada a paratireoidectomia. Na DCCP HCFMUSP, a cirurgia de escolha é a paratireoidectomia total com auto-implante imediato de fragmentos de paratireoide em membro superior. Nestes casos, caso haja recidiva do hiperparatireoidismo, pode ser causada pelo tecido implantado e, em alguns casos, há necessidade de ressecção dos implantes. Objetivo: O estudo atual tem como objetivo avaliar os pacientes submetidos a este tratamento e identificar fatores clínicos que podem prever recidiva atribuída ao implante. Método: Analisamos retrospectivamente dados de 57 doentes e identificamos nove recidivas do hiperparatireoidismo atribuídas ao implante, que foram submetidos a 14 procedimentos de retirada. Escolhemos um grupo controle (pareado por sexo e idade) de pacientes submetidos a paratireoidectomia total com auto-implante, sem recidiva por pelo menos dois anos. Resultados: Analisamos dados clínicos e laboratoriais, assim como pesos e padrões histológicos das paratireoides operadas. Houve diferença estatisticamente significante entre a dosagem pré-operatória de PTH (p=0,0091), tamanho da paratireoide ressecada durante a primeira cirurgia (p=0,0052) e padrão nodular na paratireoide selecionada para implante (p=0,0152) e recidiva no implante. Oito dos nove pacientes diagnosticados com recidiva tiveram controle da doença após os procedimentos. Todos os dados estatisticamente significativos foram pré ou peri-operatórios, e podem ajudar em decisões intra-operatórias. Conclusões: Baseado no estudo, sugerimos que a quantidade de tecido paratireoideo necessário a ser implantado pode variar conforme a dosagem sérica de PTH, o tamanho das glândulas e a presença de hiperplasia nodular no intra-operatório. As glândulas maiores e com hiperplasia nodular devem ser evitadas para implante.

Introduction: When clinical treatment of secondary hyperparathyroidism fails, parathyroidectomy is mandatory. Total parathyroidectomy and immediate parathyroid autotransplantation in the forearm is the standard treatment at the DCCP HCFMUSP. In these cases, if hyperparathyroidism reccurs, it may be caused by hyperplastic graft tissue and, sometimes, reintervention is needed to resect hyperplasic implants. Objective: The present study seeks to evaluate patients submitted to this treatment and try to clarify clinical factors that could predict hyperparathyroidism recurrence due to graft hyperplasia. Method: We could retrospectively analyse data from 57 patients, and identify nine recurrences of hyperparathyroidism caused by graft hyperplasia, submitted to 14 implant resections. We choose a control group: autotransplantation without recurrence for at least two years (matched by age and sex). Results: We analyzed pre-operative clinical and laboratorial data, parathyroid weight and histological patterns. There were statistically significant differences among preoperative serum PTH (p=0,0091), parathyroid gland size resected during first surgery (p=0,0052) and nodular pattern at parathyroid chosen for transplantation (p=0,0152) and recurrence caused by graft tissue. In all patients, but one, the disease is controlled. All statistically significant identified risk factors were pre or perioperative and may help intra operative decisions. Conclusion: Based on this study, we suggest that the amount of graft tissue may be influenced by pre-op PTH levels, size of glands and presence of nodular hyperplasia. A large and/or nodular glands should be avoided for implants.

Increased expression of monocarboxylate transporters 1, 2, and 4 in colorectal carcinomas.

Tumour cells are known to be highly glycolytic, thus producing high amounts of lactic acid. Monocarboxylate transporters (MCTs), by promoting the efflux of the accumulating acids, constitute one of the most important mechanisms in the maintenance of tumour intracellular pH. Since data concerning MCT expression in colorectal carcinomas (CRC) are scarce and controversial, the present study aimed to assess the expressions of MCT1, 2, and 4 in a well characterized series of CRC and assess their role in CRC carcinogenesis. CRC samples (126 cases) were analyzed for MCT1, MCT2, and MCT4 immunoexpression and findings correlated with clinico-pathological parameters. Expression of all MCT isoforms in tumour cells was significantly increased when compared to adjacent normal epithelium. Remarkably, there was a significant gain of membrane expression for MCT1 and MCT4 and loss of plasma membrane expression for MCT2 in tumour cells. Plasma membrane expression of MCT1 was directly related to the presence of vascular invasion. This is the larger study on MCT expression in CRC and evaluates for the first time its clinico-pathological significance. The increased expression of these transporters suggests an important role in CRC, which might justify their use, especially MCT1 and MCT4, as targets in CRC drug therapy.

Peritumoural, but not intratumoural, lymphatic vessel density and invasion correlate with colorectal carcinoma poor-outcome markers.

To evaluate whether lymphatic vessel density (LVD) and lymphatic vessel invasion (LVI) are useful markers of worse outcome in colorectal carcinoma and if LVD and LVI correlate to the classical clinical-pathological parameters, we analysed 120 cases of colorectal carcinomas selected from the files of Division of Pathology, Hospital das Clinicas, São Paulo University, Brazil. Assessment of LVD and LVI was performed by immunohistochemical detection of lymphatic vessels, using the monoclonal antibody D2-40. Higher LVD was found in the intratumoural area, when comparing with normal and peritumoural areas (p < 0.001). However, peritumoural LVD, but not intratumoural, correlated with both colonic-wall-invasion depth (p = 0.037) and liver metastasis (p = 0.012). Remarkably, LVI was found associated with local invasion (p = 0.016), nodal metastasis (p = 0.022) and hepatic metastasis (p < 0.001). Peritumoural LVD and LVI are directly related to histopathological variables indicative of poor outcome such as lymph-node status and liver metastasis.

Clinicopathologic and immunohistochemistry characterization of synchronous multiple primary gastric adenocarcinoma.

The aim of this investigation was to evaluate clinicopathologic and immunohistochemical characteristics of synchronous primary gastric adenocarcinomas. Immunohistochemistry for p53 (suppressor pathway) and for hMLH1, hMSH2, and hMSH6 (mutator pathway) was performed using ABC-technique amplification by biotinylated tyramide. Synchronous primary gastric adenocarcinomas were detected in 19/553 (3.43%) of the patients. The tumors were localized in distal stomach in 22, body in 14, and proximal in five. There was a predominance of intestinal type in the group of synchronic tumors compared to the solitary lesions, 73.2 vs 37.3%, p = 0.001. Synchronous neoplasias were diagnosed in earlier stage than solitary neoplasias, T1-T2 = 60.9% vs T1-T2 = 28.4%, p = 0.0001; and N0 = 68.4% vs N0 = 26.2%, p = 0.001. p53 was detected in 52.6% of the patients with synchronous tumors. Altered hMLH1 immunoexpression occurred in 26.3% of the patients and hMSH6 in 5.3%. hMSH2 immunoreactivity was positive in all tumors. p53 was solely detected in 17 tumors, while hMLH1 was altered in 10/24 negative p53 tumors, p = 0.01. Synchronous gastric adenocarcinomas presented higher frequency of intestinal type and early gastric cancer in comparison to solitary gastric cancer. Two routes of carcinogenesis, mutator, and suppressor appear to be involved independently in the development of synchronous tumors.

hMLH1, hMSH2 and cyclooxygenase-2 (cox-2) in sporadic colorectal polyps.

BACKGROUND: Colorectal adenomatous polyps are known as premalignant lesions. Mutations in the mismatch repair (MMR) enzymes hMLH1, hMSH2 and hMSH6 are recognized causes of hereditary non-polyposis colorectal cancer and act by inducing a mutator phenotype characterized by microsatellite instability (MSI). MSI is also detected in sporadic colorectal cancers. Cox-2 is an inducible enzyme that regulates prostaglandin synthesis and it is overexpressed at sites of inflammation, in colorectal adenomatous polyps and cancer. The aim of this study was to evaluate the immunoexpression of hMLH1, hMSH2 and Cox-2 in polyps resected through colonoscopy, and to examine their association with clinicopathological characteristics (age, gender, location, size, histology and grade of dysplasia). PATIENTS AND METHODS: One hundred and sixty-seven colonic polyps, 6 normal colonic mucosa samples, and 23 samples of colorectal adenocarcinoma were used in this study. All patients had no family history of colorectal cancer. The samples were prospectively collected and immunostained for hMLH1, hMSH2 and Cox-2 using the ABC-immunohistochemistry technique with amplification by biotinylated tyramide. The mean age was 60.2+/-13.8 years (range 21-90 years) and 77 (55.8%) were men. RESULTS: Tubular adenomas were present in 81.4%, tubulous-villous in 15.9%, serrated in 1.8%, and villous in 0.9%. The majority of the adenomas were located in the rectosigmoid region (63.5%), followed by ascendant in 14.2%, cecum in 7.5%, descendent in 8.2% and transverse in 6.7%. Low-grade dysplasia was detected in 59.6% of the adenomas. Loss of hMLH1 and hMLH2 immunoexpression was observed in 20% and 15.5% of the adenomas, respectively. Cox-2 expression was found in 9% of the adenomas, and in 40% of the adenocarcinomas. Moreover, Cox-2 immunoexpression was associated with the multiplicity of adenomas in the same patient (p=0.001). There was no association between marker immunoexpression and gender, age, location, size, histology or grade of dysplasia. CONCLUSION: Loss of hMLH1 and hMLH2 immunoexpression in adenomas is relatively frequent in patients without colorectal cancer family history. Cox-2 is overexpressed in colorectal adenomatous polyps and adenocarcinomas, and its positivity in adenomas may indicate a higher risk for multiple lesions.