ID1 and ID4 Are Biomarkers of Tumor Aggressiveness and Poor Outcome in Immunophenotypes of Breast Cancer.

Inhibitor of differentiation (ID) proteins are a family of transcription factors that contribute to maintaining proliferation during embryogenesis as they avoid cell differentiation. Afterward, their expression is mainly silenced, but their reactivation and contribution to tumor development have been suggested. In breast cancer (BC), the overexpression of ID1 has been previously described. However, whether the remaining ID genes have a specific role in this neoplasia is still unclear. We studied the mRNA expression of all ID genes by q RT-PCR in BC cell lines and 307 breast carcinomas, including all BC subtypes. Our results showed that ID genes are highly expressed in all cell lines tested. However, ID4 presented higher expression in BC cell lines compared to a healthy breast epithelium cell line. In accordance, ID1 and ID4 were predominantly overexpressed in Triple-Negative and HER2-enriched samples. Moreover, high levels of both genes were associated with larger tumor size, histological grade 3, necrosis and vascular invasion, and poorer patients' outcomes. In conclusion, ID1 and ID4 may act as biomarkers of tumor aggressiveness and worse prognosis in breast cancer, and they could be used as potential targets for new treatments discover.

Low activation of Insulin-like Growth Factor 1-Receptor (IGF1R) is associated with local recurrence in early breast carcinoma.

Background The predictive value of IGF1R on local recurrence in invasive breast carcinoma (BC) is not well known. Methods In a series of 197 lymph-node negative BC patients treated with breast-conserving surgery and radiation therapy, we performed immunohistochemistry for alpha-IGF1R, beta-IGF1R (phosphorylated/active form) and Estrogen/Progesterone receptors. We further evaluated the IGF1R mRNA expression by quantitative RT-PCR and IGF1R mutations by direct DNA sequencing (exons 19 and 21) in 85 primary BC (42 control cases, 31 with local recurrence and 12 with distant metastasis) and in 31 local recurrences. Unconditional logistic regression analyses were performed to identify risk factors for recurrence. Results Local recurrences were associated with high-grade tumors, PR-negative and low active-IGF1R, which emerged as independent breast relapse predictors by multivariate analysis. Conclusion Patients with early BC treated with lumpectomy and radiation who have low-grade tumors and favorable markers (increased content of active IGF1R and PR-positive) have a low risk of local recurrence. Therefore, do not benefit from a boost dose on the surgical scar.

Pseudoangiosarcomatous features in medullary thyroid carcinoma spindle-cell variant. Report of a case studied by FNA and immunohistochemistry.

We report a case of medullary thyroid carcinoma (MTC), spindle cell variant, which exhibited striking histological pseudovascular clefts and abortive lumina, closely mimicking an angiosarcoma. The patient is a 72-yr-old-man who presented facial rash and a solid nodule in the right lobe of the thyroid gland. The fine-needle aspiration (FNA) showed bloody background containing loosely groups of fusiform and plasmocytoid cells with coarse chromatin and eosinophilic granular cytoplasms. Microscopically, the tumor exhibited spindle-cell pattern intermingled with a striking angiosarcoma-like pattern characterized by the presence of abortive lumen and clefts containing erythrocytes. Dense hyaline extracellular amyloid was present. The tumor cells were strongly positive for cytokeratin, chromogranine-A, synaptophysine, serotonin, calcitonin, and CEA. TTF-1 was weakly positive. Stains for CD34 and CD31 were negative. This case illustrates that the spindle cell variant of MTC may exhibit an infrequent angiosarcoma-like appearance which may be misdiagnosed as angiosarcoma.

Detection of BRAF V600E mutation in colorectal cancer: comparison of automatic sequencing and real-time chemistry methodology.

Mutation V600E of BRAF, a kinase-encoding gene from the RAS/RAF/MAPK pathway, in colorectal carcinoma (CRC) suggests a sporadic origin of the disease, providing an exclusion criterion for hereditary nonpolyposis colorectal cancer. Here we describe detection of this mutation by real-time chemistry TaqMan MGB probes, confirmed by direct DNA sequencing as the gold standard. DNA was extracted from paraffin-embedded tissue from 112 tumors obtained from the EPICOLON study. Seventy-two tumors were CRC with defective DNA mismatch repair (MMR; microsatellite instability and/or loss of protein expression by immunohistochemical analysis), and 40 were proficient MMR controls. BRAF mutation was detected in 20/72 (27.8%) CRC with defective MMR and in 3/40 (7.5%) proficient MMR controls (P = 0.011). BRAF mutation was detected in 19/51 (37.3%) tumors with loss of MLH1 expression and in none of the tumors with loss of MSH2 expression (0/13). BRAF mutation was not found in cases with germline mutation of MLH1 (4/112) or MSH2 (3/112) genes. The sensitivity and specificity of our real-time chemistry were both 100% for detecting the V600E mutation. Because real-time chemistry methodology has advantages in cost, time, and labor, we consider it a valuable alternative to automatic direct sequencing, particularly for serial measurements.

Immunohistochemical analysis of tumor angiogenic factors in human pituitary adenomas.

Microvessel density (MVD) has been studied in a number of neoplasias, and apparently, there is a relationship between angiogenesis and tumor progression, response to treatment, and outcome. In pituitary adenoma, the association between MVD and vascular endothelial growth factor (VEGF) with tumor behavior has been described, but correlation with other angiogenic factors such as fetal liver kinase 1 (Flk-1) or proliferative markers is unknown. We investigated MVD, VEGF, and its receptor Flk-1 expression in 60 human pituitary adenomas: 13 growth hormone cell adenomas, 7 prolactin cell adenomas, 5 corticotroph cell adenomas, 2 thyrotroph cell adenomas, and 33 nonfunctioning adenomas (30 gonadotroph cell adenomas and 3 null cell adenomas). We performed immunohistochemistry for CD34, Ki-67, VEGF, and Flk-1. To evaluate MVD, we used 2 methods: the number of vessels per square millimeter and the Chalkley method. Immunohistochemistry results were correlated, as well as with clinicopathologic factors. Adenomas with higher MVD were thyrotroph cell adenomas (299.9 +/- 87.5), and those with lower MVD were prolactin cell adenomas (168.6 +/- 63.3; P = .45, analysis of variance). We found a trend toward higher MVD in the adenomas of older patients (P = .142), but no difference was found regarding sex, extrasellar extension, or Ki-67 (P > .05). However, extrasellar extension was nearly significant when the Chalkley method score was high (P = .056). Low expression of VEGF was seen predominantly in prolactin cell adenomas, and high in nonfunctioning adenomas, or in cases of older patients (P < or = .032). Flk-1 score correlated with VEGF (P = .006). High expression was observed in nonfunctioning adenomas, cases presenting at older ages, and with extrasellar extension (P < or = .022). Our study shows that VEGF and Flk-1 are widely expressed in pituitary adenomas, predominantly in nonfunctioning adenomas and those presenting at older ages. Moreover, Flk-1 is associated with a more aggressive phenotype, and it may have potential therapeutic interest.

Secretory carcinoma of the male breast: correlation of aspiration cytology and pathology.

Secretory carcinoma (SC) is a rare variant of breast carcinoma, which was first described in children and adolescents but it can occur at all ages. Very few cases have been reported in male patients. We describe the cytological and histopathological features of SC in a 79-yr-old man. Cytological findings demonstrated cohesive sheets of monotonous cells with round nuclei and small nucleoli. Most cells contained intracytoplasmic vacuoles, which are the key feature of an accurate diagnosis. Differential diagnosis with other tumors is discussed briefly.

Defective mismatch-repair colorectal cancer: clinicopathologic characteristics and usefulness of immunohistochemical analysis for diagnosis.

The purpose of our study was to determine the usefulness of immunohistochemical analysis for the diagnosis of mismatch-repair (MMR) gene defective colorectal tumors and to describe their prevalence and clinicopathologic characteristics. We studied 172 cases. DNA was extracted from formalin-fixed, paraffin-embedded surgical samples, and microsatellite analysis was performed by polymerase chain reaction with BAT-26. The results were correlated with immunohistochemical analysis for hMLH1 and hMSH2. Microsatellite instability (MSI) was detected in 13 (7.6%) tumors, and all showed loss of protein expression of hMLH1 (11/13) or hMSH2 (2/13) (P < .000). Patients with MMR-defective tumors more frequently had poorly differentiated tumors (5/13 [38%] vs 18/159 [11.3%]; P = .02) located in the ascending colon (8/13 [62%] vs 30/159 [18.9%]; P < .0001) and a personal history of other neoplasms (4/13 [31%] vs 18/159 [11.3%]; P = .05). There were no differences in age, family history of cancer, or TNM stage. Immunohistochemical analysis seems to be a reliable method to detect most colorectal cancers with defective MMR genes.

Organizing pneumonia adjacent to lung cancer: frequency and clinico-pathologic features.

In order to assess the frequency of peripheral organizing pneumonia (OP) in patients with resected lung tumours and to describe its differential features, a cross-sectional study with prospective data collection was realized in a community teaching hospital. Demographic and clinical data were collected from clinical records. The lung specimens removed with a curative purpose in 89 consecutive patients with lung tumours were studied and the clinical and pathological characteristics of patients with and without OP were compared. In 33 of 89 patients (37%) included, OP in the vicinity of neoplasm was found. Areas of other types of fibrosis were evident in 21 patients (24%). Male gender, smoker, epidermoid histological type and the presence of lipid pneumonia were found with a significant higher frequency in patients with OP. Although without significant differences, the presence of symptoms and the bronchial stenosis were found more frequently in patients with OP. In conclusion, OP pattern adjacent to lung cancer, frequently associated to lipid pneumonia, is a common pathological finding. Male gender, a history of tobacco use and epidermoid histological type appear as risk factors for developing this pathologic pattern. Given the lack of distinctive clinico-pathological features, cancer adjacent OP could be confounded with other etiologic forms of this fibrotic process.

Angiogenic Index: A New Method for Assessing Microvascularity in Breast Carcinoma with Possible Prognostic Implications.

The quantitation of microvessels in breast cancer in sections immunolabeled for factor VIII, CD-34, or CD-31 currently is expressed as numbers of vessels/mm2. In an attempt to perform a more accurate method for counting microvessels, we determined the number of microvessels by 1,000 tumor cells (angiogenic index; AI). This new method minimizes the possible variations concerning the width of the microscopic fields, stroma/epithelium relations, and cellular tumor size. The present study compares the number of vessels determined by the traditional method (vessels/mm2) and AI in a series of 215 cases of ductal infiltrating carcinoma not otherwise specified. Also we studied the degree of correlation between both methods and with pathologic variables (tumor size, histologic grade, mitotic count, tumor necrosis, vascular invasion, skin infiltration, and axillary lymph node metastasis). Our results showed that the AI correlates more significantly than the classic microvessel density determination with other prognostic factors in breast cancer. In histologic grade III tumors, high AI correlates significantly with the presence of more than three axillary lymph nodes metastases. Therefore we recommend determining the degree of tumor angiogenesis by counting vessels per 1,000 tumor cells (AI) because of its reliable determination and significant correlation with other prognostic factors.