B-cell lymphoma-2 receptor in human primary breast and its lymph node metastases: more than a surrogate marker

Background: Due to its anti-apoptotic and anti-proliferative contradictory functions, BCL2 role in breast carcinoma progression is not clearly understood. The purpose of this study was to highlight BCL2 expression during metastatic progression of invasive breast carcinoma of no special type (NST). Materials and methods: The specimens, primary tumors and corresponding lymph node metastases (LNM) from 84 patients were immunostained for estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor (HER)-2, basal cytokeratin CK5, nuclear protein Ki67 and B-cell lymphoma (Bcl)-2 receptor. Results: BCL2 expression was higher at primary site than in axillary metastases. Its score correlates positively with hormone receptors’ level and negatively with HER2, CK5 and Ki67 at both sites. Switch of molecular profile was determined in 22.62% of cases. BCL2 expression was not influenced by subtypes switch. Changes of BCL2 expression were found in 25% of cases with stable molecular subtype. The Luminal A and Luminal B/Ki67 were encountered in the majority of BCL2 transitions, mainly from positive to negative state. Conclusions: Molecular subtypes and BCL2 expression are not stable during tumor progression and metastatic development. In the present study we established immunohistochemically that BCL2 is not influenced by subtypes’ transitions. BCL2 switches were encountered only in cases with a stable HER2, Luminal A or B phenotypes. We expect a further confirmation of our results by other research groups. Key words: BCL2, breast carcinoma, immunohistochemistry, molecular subtypes, metastases

Oddziaływanie kwaśnego białka włókienkowego gleju i białka S100 w gruczolakach przysadki mózgowej: dwóch czy więcej graczy?

Introduction: S100 protein and GFAP expression in pituitary adenomas tumour cells is not well known; few correlations with other prognostic or therapeutic factors have previously been reported in pituitary adenomas. We aim to elucidate their involvement in the pathogenesis of pituitary adenomas and to establish the correlation of their expression with different growth factors and growth factor receptors known to have a prognostic and/or therapeutic role. Material and methods: Sixty-one cases of pituitary adenomas were immunohistochemically assessed for the expression of GFAP and S100 protein in both tumour cells and FS cells, in close relationship with hormone profile, and correlated with vascular endothelial growth factor (VEGF) and epidermal growth factor receptor (EGFR) expression, previously studied by our team. Results: GFAP and S100 protein were expressed both in tumour cells and FS cells. Differences between morphology, distribution, and density of GFAP+ FS cells and S100+ FS cells were observed according to the hormone profile of pituitary adenomas. GFAP and S100 protein expression in tumour cells was significantly related to hormone profile of pituitary adenomas and also with VEGF and EGFR expression. Conclusions: GFAP and S100 protein expressions in tumour cells from pituitary adenomas are influenced by hormone profile. Our re­sults support the presence of two molecular subtypes of FS cells GFAP+/VEGF+/S100 respectively and another one that is GFAP-/S100+/EGFR+ simultaneously with the classical variant GFAP+/S100+. It is possible that S100+/EGFR+ pituitary adenomas represent a group of pituitary adenomas with an aggressive behaviour and a high ability of invasion and recurrence.Wstęp: Ekspresja białka S100 i kwaśnego białka włókienkowego gleju (GFAP, glial fibrillary acid protein) w gruczolakach przysadki mózgowej nie została dobrze poznana. Dostępne są nieliczne publikacje dotyczące korelacji tych cząsteczek z innymi czynnikami prognostycznymi lub terapeutycznymi w gruczolakach przysadki. Celem niniejszej pracy jest wyjaśnienie udziału tych białek w patogenezie gruczolaków przysadki i ustalenie korelacji między ich ekspresją a różnymi czynnikami wzrostu lub receptorami czynników wzrostu o znanej wartości prognostycznej i/lub terapeutycznej. Materiał i metody: Sześćdziesiąt sześć przypadków gruczolaka przysadki zbadano metodami immunohistochemicznymi w celu oceny ekspresji białek GFAP i S100 zarówno w komórkach guza, jak i w komórkach pęcherzykowo-gwiaździstych (FS, folliculo-stellate cells) oraz przeanalizowania uzyskanych wyników w ścisłej zależności z profilami hormonalnymi gruczolaków i w korelacji z ekspresją czynnika wzrostu śródbłonka naczyniowego (VEGF, vascular endothelial growth factor) i receptora naskórkowego czynnika wzrostu (EGFR, epidermal growth factor receptor), ocenianych w badaniu przeprowadzonym wcześniej przez nasz zespół. Wyniki: Ekspresję białek GFAP i S100 stwierdzono zarówno w komórkach guza, jak i w komórkach FS. Zaobserwowano różnice pod względem morfologii, rozkładu i gęstości komórek FS z dodatnią ekspresją białek GFAP i S100 (GFAP+FS i S100+FS) odpowiadające profilowi hormonalnemu gruczolaków przysadki. Ekspresja białek GFAP i S100 w komórkach guza była istotnie związana z profilem hormonalnym gruczolaków przysadki, a także z ekspresją VEGF i EGFR. Wnioski: Ekspresja białek GFAP i S100 w komórkach gruczolaka przysadki zależy od profilu hormonalnego guza. Uzyskane w badaniu wyniki potwierdzają obecność dwóch podtypów molekularnych komórek FS GFAP+/VEGF+/S100 oraz jednego typu GFAP–/S100+/EGFR+ występujących jednocześnie z klasycznym wariantem GFAP+/S100+. Możliwe, że gruczolaki przysadki z ekspresją S100+/EGFR+ należą do grupy tych nowotworów cechujących się agresywnością i dużą zdolnością do naciekania i nawrotów

Toward a Molecular Classification of the Head and Neck Squamous Cell Carcinoma

The frequency of the squamous cell carcinoma of the head and neck is constantly increasing, with over 550.000 new cases registered globally each year. The conventional histopathological diagnosis most commonly indicates the squamous cell carcinoma as tumor type and G2 as differentiation grade. Despite of this relative morphological uniformity, there is a great heterogeneity in the molecular profile, the therapeutic response and prognosis. Most probably, this entity includes many diseases, similar in basic morphologic features, but different in the biological behavior. Trying to answer this question and to show discrepancies when they exist, we have evaluated in this book chapter, our own results and data from the literature in terms of molecular profile at the protein level, including the spectrum of proliferation markers, growth factors and their receptors, stromal proliferation, angiogenesis and lymphangiogenesis. These data will allow to identify some major criteria for a better stratification of cases, selected for gene analysis and personalized therapy as a future perspective and direction

Inhibition of Notch signaling induces extensive intussusceptive neo-angiogenesis by recruitment of mononuclear cells

Notch is an intercellular signaling pathway related mainly to sprouting neo-angiogenesis. The objective of our study was to evaluate the angiogenic mechanisms involved in the vascular augmentation (sprouting/intussusception) after Notch inhibition within perfused vascular beds using the chick area vasculosa and MxCreNotch1(lox/lox) mice. In vivo monitoring combined with morphological investigations demonstrated that inhibition of Notch signaling within perfused vascular beds remarkably induced intussusceptive angiogenesis (IA) with resultant dense immature capillary plexuses. The latter were characterized by 40% increase in vascular density, pericyte detachment, enhanced vessel permeability, as well as recruitment and extravasation of mononuclear cells into the incipient transluminal pillars (quintessence of IA). Combination of Notch inhibition with injection of bone marrow-derived mononuclear cells dramatically enhanced IA with 80% increase in vascular density and pillar number augmentation by 420%. Additionally, there was down-regulation of ephrinB2 mRNA levels consequent to Notch inhibition. Inhibition of ephrinB2 or EphB4 signaling induced some pericyte detachment and resulted in up-regulation of VEGFRs but with neither an angiogenic response nor recruitment of mononuclear cells. Notably, Tie-2 receptor was down-regulated, and the chemotactic factors SDF-1/CXCR4 were up-regulated only due to the Notch inhibition. Disruption of Notch signaling at the fronts of developing vessels generally results in massive sprouting. On the contrary, in the already existing vascular beds, down-regulation of Notch signaling triggered rapid augmentation of the vasculature predominantly by IA. Notch inhibition disturbed vessel stability and led to pericyte detachment followed by extravasation of mononuclear cells. The mononuclear cells contributed to formation of transluminal pillars with sustained IA resulting in a dense vascular plexus without concomitant vascular remodeling and maturatio

The multifaceted role of podoplanin expression in hepatocellular carcinoma

The role of podoplanin in hepatocellular carcinoma (HCC) is not clear yet. The aim of our study was to evaluate the expression of podoplanin in HCC and to determine its role in hepatocarcinogenesis. We performed immunohistochemistry with monoclonal D2-40 antibody, on paraffin-embedded tissue sections of 72 patients diagnosed with HCC. Lymphatic vessels density (LVD) was increased in patients who had vascular invasion at the time of diagnosis (P=0.018) and in those with associated cirrhosis (P=0.006). Tumor cells showing podoplanin expression were correlated with histological grade (P=0.040). Podoplanin-expressing cancer associated fibroblasts (CAFs) were correlated with both LVD (P=0.019) and tumor cells (P=0.015). Our results sustain the dual role of podoplanin in HCC by its involvement in both HCC tumorigenesis, lymphatic neovascularization and tumor invasion invasiveness. A possible crosstalk between epithelial and stromal tumor cells in HCC tumor microenvironment may be mediated by podoplanin, but this hypothesis needs further studies to elucidate this interrelation.</p

Proliferating Lymphatic Endothelial Cells as a Prognostic Marker in Head and Neck Squamous Cell Carcinoma

Podoplanin and Ki-67 are two important markers of cancer progression. The aim of this study is to evaluate double immunostaining for Ki-67 and podoplanin in head and neck squamous cell carcinoma (HNSCC), and to observe the involvement of lymphagiogenesis in tumoral and peritumoral areas, as well as the density of tumor proliferation correlated with histopathological grading. A total of 50 patients with HNSCC were included in this study. We carried out a morphological evaluation of tissue samples, after that, cases were selected for double Ki-67 and podoplanin immunostaining. Podoplanin expression was significantly correlated with histopathological grade (p p = 0.037) and expression of Ki-67 (p p = 0.050). A high expression of podoplanin, as well as of the proliferation factor Ki-67, was observed in histopathological grade G3 and the correlation between these (p p = 0.028), and implication of LMVD and LVI was not significant (LMVD p = 0.577; LVI p = 0.976). This study demonstrated the importance of double immunolabeling in assessing lymphagiogenesis and tumor proliferation in correlation with histopathological grades in HNSCC

Chloride Intracellular Channel Protein 1 Expression and Angiogenic Profile of Liver Metastasis of Digestive Origin

Chloride intracellular channel 1 (CLIC1) is involved in cell migration and metastasis. The histological growth patterns of liver metastasis are as follows: desmoplastic (d-HGP), replacement (r-HGP), pushing (p-HGP), and mixed. The aim of this study was to evaluate the relation between HGP, angiogenesis, and CLIC1 expression. Materials and Methods: A total of 40 cases of primary tumors and their LM: d-HGP (12 cases), r-HGP (13 cases), and p-HGP (15 cases), were evaluated through simple and double immunostaining. CLIC1 assessment was conducted as follows: scores of 0 (less than 10% of positive cells), 1 (10–30%), 2 (30–50%), or 3 (more than 50%) were assigned. Heterogeneous CLIC1 expression was found. CLIC1 in primary tumors correlated with grade G for all cases of LM with a p-HGP (p = 0.004). The CLIC1 score for LMs with an r-HGP correlated with grade G of the corresponding primary tumor (p = 0.027). CLIC1 and CD34+/Ki67+ vessels (p = 0.006) correlated in primary tumors. CLIC1 in primary tumors correlated with CD34+/Ki67+ vessels of LMs with a d HGP (p = 0.024). Conclusions: The CLIC1 score may have prognostic value, mainly for LMs with a p-HGP and r-HGP, and therapeutic value for LMs with a d-HGP

Novel Expression of Thymine Dimers in Renal Cell Carcinoma, Demonstrated through Immunohistochemistry

Despite significant developments in renal cell carcinoma (RCC) detection and molecular pathology, mortality has been steadily rising. Advanced RCC remains an incurable disease. Better clinical management tools, i.e., RCC biomarkers, have yet to emerge. Thymine-dimers (TDs) were traditionally considered photo-dependent pre-mutagenic lesions, occurring exclusively during ultra-violet light exposure. Non-oxidative, direct, and preferential byproducts of DNA photochemical reactions, TDs, have recently shown evidence regarding UVR-independent formation. In this study, we investigate, for the first time, TD expression within RCC tumor tissue and tumor-adjacent healthy renal parenchyma using a TD-targeted IHC monoclonal antibody, clone KTM53. Remarkably, out of the 54 RCCs evaluated, 77.8% showed nuclear TD-expression in RCC tumor tissue and 37% in the tumor-adjacent healthy renal parenchyma. A comprehensive report regarding quantitative/qualitative TD-targeted immunostaining was elaborated. Two main distribution models for TD expression within RCC tumor tissue were identified. Statistical analysis showed significant yet moderate correlations regarding TD-positivity in RCC tissue/tumor-adjacent healthy renal parenchyma and TNM stage at diagnosis/lymphatic dissemination, respectively, indicating possible prognostic relevance. We review possible explanations for UVR-independent TD formation and molecular implications regarding RCC carcinogenesis. Further rigorous molecular analysis is required in order to fully comprehend/validate the biological significance of this newly documented TD expression in RCC