Expression Of Basal Markers Research Articles

P0067 Basal cell markers for differentiation of prostate adenocarcinoma from cancer mimickers

<h3>Background</h3> Prostate cancer is the leading cause of cancer and the second cause of cancer-related death in men after lung cancer in the world. Differentiation of prostate adenocarcinoma from benign prostate lesions and hyperplasia sometimes cannot be done on the basis of morphological findings. As there is no basal cell layer in prostate adenocarcinoma, basal cell markers could help to differentiate prostate adenocarcioma from cancer mimickers. <h3>Methods</h3> We studied 98 prostate biopsies (40 adenocarcinoma and 58 benign lesions) for expression of basal cell markers. <h3>Findings</h3> P63 and 34<i>β</i>E12 were negative in all prostate adenocarcinoma specimens, but all benign prostate hyperplasia and high grade intraepithelial neoplasia cases expressed them. <h3>Interpretation</h3> Basal cell markers can help to distinguish prostate adenocarcioma from cancer mimickers.

Macrophage expression of activation markers in iron deficient conditions (1034.18)

Objective: To determine the expression of major histocompatibility complex (MHC) class I and II and costimulatory molecules (CD80/CD86) by macrophages from iron deficient mice.Methods: Two groups of BALB/c mice were established from gestation: control (standard diet, 100ppm iron) and iron deficient (iron deficient diet, 10ppm iron). At 21 days of age, pups were weaned and their spleens were dissected for mononuclear cell purification. Purified cells were stained with fluorochrome‐conjugated antibodies for the surface markers CD11b, CD45, F4.80, MHC‐I, MHC‐II, CD80 and CD86. Analysis of expression of these markers was performed by flow cytometry.Results: The percentage of positive cells for MHC‐I, CD80 and CD86 was higher in iron deficient mice compared to controls. In contrast, the percentage of positive cells for MHC‐I was lower in iron deficient mice. However, the number of cells expressing MHC‐I, CD80 and CD86 was higher in the iron deficient group, while the number of cells expressing MHC‐II was higher in the control group. The mean fluorescence intensity for all markers was higher in iron deficient mice.Conclusion: Iron deficiency increases the basal expression of cell surface markers of activation in splenic macrophages. These results may have important implications in the activation of leucocytes in basal conditions in iron deficient individuals.

MP19-17 SURFACE CD24 DISTINGUISHES BETWEEN LOW DIFFERENTIATED AND TRANSIT-AMPLIFYING CELLS IN THE BASAL LAYER OF THE HUMAN PROSTATE

You have accessJournal of UrologyBenign Prostatic Hyperplasia: Basic Research1 Apr 2014MP19-17 SURFACE CD24 DISTINGUISHES BETWEEN LOW DIFFERENTIATED AND TRANSIT-AMPLIFYING CELLS IN THE BASAL LAYER OF THE HUMAN PROSTATE Christian Schwentner, Neli Petkova, Jörg Hennenlotter, Tilman Todenhöfer, Hans-Joerg Buehring, Stefan Aufderklamm, and Arnuf Stenzl Christian SchwentnerChristian Schwentner More articles by this author , Neli PetkovaNeli Petkova More articles by this author , Jörg HennenlotterJörg Hennenlotter More articles by this author , Tilman TodenhöferTilman Todenhöfer More articles by this author , Hans-Joerg BuehringHans-Joerg Buehring More articles by this author , Stefan AufderklammStefan Aufderklamm More articles by this author , and Arnuf StenzlArnuf Stenzl More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2014.02.718AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Benign prostatic hyperplasia (BPH) and prostate cancer (PCa) are common abnormalities in elderly men. It is considered that epithelial stem cells are involved in the etiology and development of both diseases. To distinguish aberrant from normal cells, the knowledge about primary epithelial stem/progenitor cells (ES/P) is essential. The aim of this study was to examine the role of surface markers to distinguish between different subsets of prostate basal epithelium. METHODS The expression pattern of prostate tissue single cell suspensions was analyzed by flow cytometry using different markers. Sorted cell populations were examined for their clonogenic capacity and the resulted colonies were analyzed with flow cytometry, Western blot, and qPCR for stem cell, basal, and luminal epithelium markers. Additionally, the histological localization of the examined markers was determined using immunofluorescence. RESULTS Using the combination of CD49f, Trop-2, and surface CD24, basal cell subsets with distinct differentiation capacities were dissected (CD49f(+) Trop-2(+) CD24(-) and CD49f(+) Trop-2(+) CD24(+) ). Although cells from the two subsets gave rise to similar basal colonies, qPCR of primary tissue revealed that higher levels of basal marker expression were detected in the CD49f(+) Trop-2(+) CD24(-) subset. Immunofluorescence analysis showed a prominent expression of CD24 by luminal and basal cells. CONCLUSIONS Subsets with distinct differentiation capacities within the basal epithelium (CD49f(+) Trop-2(+) CD24(-) and CD49f(+) Trop-2(+) CD24(+) ) can be distinguished in human prostate. CD24 is a marker expressed on the basal transit-amplifying cells (transition cells) and may play a role in the differentiation and migration of ES/P cells to the luminal layer. The knowledge of this mechanism is of relevance for treatment of both diseases. © 2014FiguresReferencesRelatedDetails Volume 191Issue 4SApril 2014Page: e195 Peer Review Report Advertisement Copyright & Permissions© 2014MetricsAuthor Information Christian Schwentner More articles by this author Neli Petkova More articles by this author Jörg Hennenlotter More articles by this author Tilman Todenhöfer More articles by this author Hans-Joerg Buehring More articles by this author Stefan Aufderklamm More articles by this author Arnuf Stenzl More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Loss of androgen receptor expression predicts early recurrence in triple-negative and basal-like breast cancer

Treatment of triple-negative invasive breast cancers, defined by the absence of estrogen and progesterone receptors and c-erbB2 expression, remains challenging. Androgen receptor, a member of the nuclear receptor superfamily that is involved in signaling pathways regulating cell proliferation, has been implicated in breast tumorigenesis. We immunohistochemically examined the expression of androgen receptor, basal markers (CK14, 34βE12) and EGFR in 699 triple-negative invasive breast cancers in tissue microarrays using the streptavidin–biotin method, and correlated the findings with clinical outcome. Positive androgen receptor expression was defined as staining of 1% or more of tumor cell nuclei. Survival outcomes were estimated with the Kaplan–Meier method and compared between groups with log-rank statistics. Cox proportional hazards models were used to determine the effect of androgen receptor on survival outcomes. Immunohistochemical positivity was observed in 38% of tumors, with the proportion of stained tumor cells ranging from 1 to 95% (mean 29%, median 10%). Androgen receptor expression was inversely associated with histologic grade and mitotic score. CK14, 34βE12 and EGFR confirmed 85% of cases to be basal-like, without significant association of basal-like phenotype with androgen receptor expression. Disease-free survival was significantly better in androgen receptor-positive triple-negative breast cancer, with a trend for improved overall survival. Decreased recurrence likelihood in both triple-negative and basal-like tumors (hazard ratio, 0.704; 95% confidence intervals, 0.498–0.994; P=0.0464; and hazard ratio, 0.675; 95% confidence intervals, 0.468–0.974; P=0.0355, respectively) was noted within 5 years of diagnosis but not thereafter. Our study suggests that loss of androgen receptor in triple-negative breast cancers augurs a worse prognosis, including those with basal-like features. More work in elucidating its relationship with mechanisms of progression, as well as trials of targeted treatment for androgen receptor-expressing triple-negative tumors, needs to be performed.

Triple-negative, Basal Marker-expressing, and High-grade Breast Carcinomas are Characterized by High Lymphatic Vessel Density and the Expression of Podoplanin in Stromal Fibroblasts

Podoplanin, expressed in the lymphatic but not in the blood vessel endothelium, is widely used as a specific marker for lymphatic endothelial cells and lymphangiogenesis. The relation between lymphatic vessel density and breast cancer subtype or the expression of basal markers has not yet been investigated. We assessed lymphatic vessel density (LVD), blood vessel density, and the expression of podoplanin in stromal cancer-associated fibroblasts, in 156 invasive ductal breast cancers (T≥1, N≥1, M0). Afterwards, we assessed the relationship between the above-mentioned parameters and (i) breast cancer subtype (luminal vs. HER2 vs. triple negative), (ii) tumor grade (G1 vs. G2 vs. G3), (iii) the expression of cytokeratin (CK) 5/6, (iv) P-cadherin, (v) smooth muscle actin (SMA), and (vi) the pattern/intensity of stromal lymphocytic infiltration. We found a significantly higher LVD and podoplanin expression in stromal fibroblasts in (i) G3 tumors, (ii) triple-negative carcinomas, (iii) tumors expressing CK5/6, SMA, or P-cadherin, and (iv) neoplasms with stroma intensively infiltrated by lymphocytes. Moreover, we observed a significant inverse relationship between the expression of podoplanin in luminal A subtype, P-cadherin, CK5/6, and SMA-negative tumors and tumors without strong lymphocytic infiltration. A significantly higher percentage of tumors with strong lymphocytic infiltration was noted among G3 carcinomas. Breast carcinomas of different grades, subtypes, and basal marker expression are characterized by different composition of the stroma, that is different LVD, podoplanin expression in stromal fibroblasts, and the pattern/intensity of lymphocytic infiltration.

Staphylococcus aureus inhibits terminal differentiation of normal human keratinocytes by stimulating interleukin-6 secretion

BackgroundStaphylococcus aureus (S. aureus) is found on the skin of approximately 90% of patients with atopic dermatitis and approximately 20% of apparently healthy subjects. S. aureus induces keratinocytes and immune cells to secrete immunoregulatory factors that cause epidermal barrier dysfunction in atopic skin. ObjectiveThis study examined factors that cause epidermal permeability barrier dysfunction in skin colonized by S. aureus. MethodsWe examined the effect of S. aureus on keratinocyte differentiation in the stratum corneum (SC) of in vivo skin, normal human keratinocytes (NHKs) and a reconstructed human epidermis (RHE) model. The fold change in expression of the terminal differentiation markers and the level of secreted cytokines were investigated. ResultsThe SC displayed decreased expression of keratin 10 (KRT 10). NHKs treated with S. aureus extracts increased expression of interleukin (IL)-6 and significantly reduced expression of the terminal differentiation markers KRT 1, KRT 10, loricrin (LOR), and filaggrin (FLG); however, the expression of basal layer markers (KRT 5, KRT 14) remained unchanged. Treatment of NHKs with an anti-IL-6 antibody in combination with IL-6 or the S. aureus extracts inhibited the decrease in KRT 10 mRNA or protein expression. After the RHEs were exposed to the S. aureus extracts, KRT 1 and KRT 10 protein levels decreased. ConclusionsThese findings suggest that S. aureus inhibits the terminal differentiation of keratinocytes by stimulating IL-6 secretion.

Myc is required for β-catenin-mediated mammary stem cell amplification and tumorigenesis

BackgroundBasal-like breast cancer is a heterogeneous disease characterized by the expression of basal cell markers, no estrogen or progesterone receptor expression and a lack of HER2 ov erexpression. Recent studies have linked activation of the Wnt/β-catenin pathway, and its downstream target, Myc, to basal-like breast cancer. Transgenic mice K5ΔNβcat previously generated by our team present a constitutive activation of Wnt/β-catenin signaling in the basal myoepithelial cell layer, resulting in focal mammary hyperplasias that progress to invasive carcinomas. Mammary lesions developed by K5ΔNβcat mice consist essentially of basal epithelial cells that, in contrast to mammary myoepithelium, do not express smooth muscle markers.MethodsMicroarray analysis was used to compare K5ΔNβcat mouse tumors to human breast tumors, mammary cancer cell lines and the tumors developed in other mouse models. Cre-Lox approach was employed to delete Myc from the mammary basal cell layer of K5ΔNβcat mice. Stem cell amplification in K5ΔNβcat mouse mammary epithelium was assessed with 3D-culture and transplantation assays.ResultsHistological and microarray analyses of the mammary lesions of K5ΔNβcat females revealed their high similarity to a subset of basal-like human breast tumors with squamous differentiation. As in human basal-like carcinomas, the Myc pathway appeared to be activated in the mammary lesions of K5ΔNβcat mice. We found that a basal cell population with stem/progenitor characteristics was amplified in K5ΔNβcat mouse preneoplastic glands. Finally, the deletion of Myc from the mammary basal layer of K5ΔNβcat mice not only abolished the regenerative capacity of basal epithelial cells, but, in addition, completely prevented the tumorigenesis.ConclusionsThese results strongly indicate that β-catenin-induced stem cell amplification and tumorigenesis rely ultimately on the Myc pathway activation and reinforce the hypothesis that basal stem/progenitor cells may be at the origin of a subset of basal-like breast tumors.

Abstract IA08: Studying therapy response and resistance in mouse models of breast cancer

Abstract Women carrying germline mutations in BRCA1 are strongly predisposed to developing triple-negative breast cancers, which frequently contain TP53 mutations. To study the role of BRCA1 loss-of-function in development of triple-negative breast cancer, we have established genetically engineered mouse models (GEMMs) for BRCA1-mutated breast cancer based on cre-mediated tissue-specific deletion of p53 and Brca1. In addition to GEMMs for Brca1-null tumors, we have generated mammary tumor models carrying specific BRCA1 founder mutations (C61G, 185delAG and 5382insC). The mammary tumors that arise in these mouse models show strong similarity to BRCA1-associated breast cancer with respect to expression of basal cell markers, high tumor grade, and lack of expression of hormone receptors and ERBB2. In addition to these GEMMs, we have also established a panel of patient-derived xenograft (PDX) models for BRCA1-proficient and –deficient triple-negative breast cancer. BRCA1 deficiency in these PDX tumors is driven either by genetic mutation or by BRCA1 promoter methylation. The BRCA1-deficient tumors that arise in our various mouse models are in all cases characterized by a high degree of genomic instability and hypersensitivity to DNA-damaging agents due to loss of homology-directed double-strand break (DSB) repair. We have successfully used our GEMMs and PDX models of BRCA1-deficient breast cancer for preclinical evaluation of therapy response and elucidation of mechanisms of acquired drug resistance. BRCA1-deficient mammary tumors are highly sensitive to DNA-damaging agents such as platinum drugs and to PARP inhibitors, which indirectly induce DSBs by inhibiting DNA single-strand break repair. However, none of these drugs are capable of causing tumor eradication and all tumors grow back after drug treatment. By employing in vitro functional genetic screens and in vivo intervention studies, we found that therapy response and resistance is affected by several factors, including type of BRCA1 founder mutation, drug efflux transporter activity, and 53BP1 status. The lack of tumor eradication prompted us to look for additional drugs targeting BRCA-deficient tumors. Using a cell-based screening approach, we found that bifunctional alkylators such as nimustine may cause durable complete remission of BRCA-deficient mouse mammary tumors, suggesting that BRCA-mutated hereditary breast cancers and BRCA-like sporadic tumors may be eradicated by dose-intensive treatment with bifunctional alkylators. In support of this notion, patients with advanced breast cancers that display a BRCA1-like profile of genomic aberrations show a high complete remission rate and long progression-free survival after treatment with high-dose alkylating chemotherapy. Citation Format: Jos Jonkers. Studying therapy response and resistance in mouse models of breast cancer. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Breast Cancer Research: Genetics, Biology, and Clinical Applications; Oct 3-6, 2013; San Diego, CA. Philadelphia (PA): AACR; Mol Cancer Res 2013;11(10 Suppl):Abstract nr IA08.

Abstract A040: Id-1 increases mammary basal stem cell activity and tumorigenesis by activating the Wnt/c-Myc pathway

Abstract Inhibitor of differentiation/DNA binding (Id)-1 is a crucial regulator of mammary development and breast cancer progression. Here, we demonstrate that Id-1 induces mammary tumorigenesis by increasing normal and malignant mammary stem cell (MaSC) activities. MaSC expansion and increased self-renewal and in vivo regenerative capacity of MaSCs were observed in the mammary glands of MMTV-Id-1 transgenic mice. Id-1 maintains the MaSC-enriched basal cells, but not the luminal cell lineage, in the mammary glands. Furthermore, Id-1 induces ductal hyperplasia and mammary tumors with high expression of basal markers in MMTV-Id-1 transgenic mice. It also increases breast cancer stem cell populations and activity in human breast cancer lines. In experiments with mouse primary mammary epithelial cells and human cancer cells, the effects of Id-1 on normal and cancer stem cell activity were shown to be mediated by the Wnt/c-Myc pathway. Collectively, these findings indicate that Id-1 regulates mammary basal stem cells by activating the Wnt/c-Myc pathway, thereby contributing to basal marker-positive breast tumor development. Citation Format: Dong-Hui Shin, Jeong-Yeon Lee, Ki-Seok Jang, Kyueng-Whan Min, Si-Hyong Jang, Seung Hyun Oh, Gu Kong. Id-1 increases mammary basal stem cell activity and tumorigenesis by activating the Wnt/c-Myc pathway. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Breast Cancer Research: Genetics, Biology, and Clinical Applications; Oct 3-6, 2013; San Diego, CA. Philadelphia (PA): AACR; Mol Cancer Res 2013;11(10 Suppl):Abstract nr A040.

Surface CD24 distinguishes between low differentiated and transit‐amplifying cells in the basal layer of human prostate

Benign prostatic hyperplasia (BPH) and prostate cancer (PCa) are common abnormalities in elderly men. It is considered that epithelial stem cells are involved in the etiology and development of both diseases. To distinguish aberrant from normal cells, the knowledge about primary epithelial stem/progenitor cells (ES/P) is essential. The aim of this study was to examine the role of surface markers to distinguish between different subsets of prostate basal epithelium. The expression pattern of prostate tissue single cell suspensions was analyzed by flow cytometry using different markers. Sorted cell populations were examined for their clonogenic capacity and the resulted colonies were analyzed with flow cytometry, Western blot, and qPCR for stem cell, basal, and luminal epithelium markers. Additionally, the histological localization of the examined markers was determined using immunofluorescence. Using the combination of CD49f, Trop-2, and surface CD24, basal cell subsets with distinct differentiation capacities were dissected (CD49f(+) Trop-2(+) CD24(-) and CD49f(+) Trop-2(+) CD24(+) ). Although cells from the two subsets gave rise to similar basal colonies, qPCR of primary tissue revealed that higher levels of basal marker expression were detected in the CD49f(+) Trop-2(+) CD24(-) subset. Immunofluorescence analysis showed a prominent expression of CD24 by luminal and basal cells. Subsets with distinct differentiation capacities within the basal epithelium (CD49f(+) Trop-2(+) CD24(-) and CD49f(+) Trop-2(+) CD24(+) ) can be distinguished in human prostate. CD24 is a marker expressed on the basal transit-amplifying cells (transition cells) and may play a role in the differentiation and migration of ES/P cells to the luminal layer. The knowledge of this mechanism is of relevance for treatment of both diseases.

Snail: a target for treating basal-like breast cancer

259 ISSN 1758-1923 10.2217/BMT.13.32 © 2013 Future Medicine Ltd Breast Cancer Manage. (2013) 2(4), 259–262 Breast cancer is a heterogeneous disease that can be divided into five biological subtypes based on gene-expression profiling: luminal A, luminal B, ErbB2, normallike and basal-like [1]. Of all the molecular subtypes, basal-like breast cancer (BLBC) is of particular clinical significance, as it associates with an aggressive clinical history, development of recurrence, distant metastasis and shorter survival, and usually occurs in younger women [2,3]. The terminology ‘basal-like’ stems from the similarity between the molecular profiling of tumors pertaining to this subgroup with that of undifferentiated breast stem cells, which reside with basal/myoepithelial cells (termed basal-like cells) of the normal breast [3]. In addition to BRCA1 germline mutations or a dysfunctional BRCA1 pathway [3], BLBC is commonly characterized by the lack of expression of estrogen receptor, progesterone receptor and HER2, and positive expression of basal markers (CK5/6 and CK14) [2,3]. The consequent absence of effective targeted therapies, such as endocrine and anti-HER2 therapies, and poor response to standard chemotherapy, which is common among patients, often results in a rapidly fatal clinical outcome for this disease. This type of breast cancer has a tendency towards early metastatic spread to the brain and lung, which are sites known to be associated with a poor prognosis and short survival [2,3]. Thus, novel treatment strategies based on the biology of BLBC are urgently needed to combat this life-threatening disease. Intriguingly, BLBC possesses many epithelial–mesenchymal transition (EMT) characteristics and exhibits robust cancer stem cell (CSC)-like features [4,5]. EMT is an essential phenotypic conversion during embryonic development, tissue remodeling, wound healing and metastasis [6,7]. In these EMT processes, epithelial cells acquire f ibroblast-like properties and exhibit reduced intercellular adhesion and increased motility; they can also break through the basal membrane and migrate over long distances, owing to profound changes in their cytoskeletal architecture. Activation of this normally latent EMT program confers a distinct advantage on cancer cells for invasion and metastasis to distant organs or tissues. In addition to enhancing the invasiveness and metastatic

Evaluation of potential predictive markers of efficacy of dacomitinib in patients (pts) with recurrent/metastatic SCCHN from a phase II trial.

6041 Background: Dacomitinib is an irreversible pan-HER TKI with preclinical (EGFRvIII+ cell lines, SCCHN xenografts) and clinical activity (phase II recurrent/metastatic SCCHN; Razak et al, Ann Oncol 2012). However, little is known about predictive markers of efficacy related to EGFR signalling in this setting. Methods: Of69 pts treated with 1st-line dacomitinib in a phase II trial for recurrent/metastatic SCCHN, 48 pts had archival tumor specimens obtained before treatment and 13 had paired biopsies (days 0 and 7 of therapy, FFPE and snap frozen). EGFRvIII and PTEN (IHC), HPV genotyping and human genomic mutations (Sequenom OncoCarta Panel – 19 genes, 238 mutations) were evaluated on archival tissue. IHC expression of AKT, CC3, EGFR, ERK, HER2, HER3, MET, Ki67, pAKT, pEGFR, pERK, pHER2 and pMET was evaluated in paired specimens. The presence/absence or expression level of these markers was correlated with response (RR)/clinical benefit (CB), PFS and OS. Results: In pts with archival tissue, no statistically significant difference was found in RR/CB or PFS based on HPV, EGFRvIII, PTEN or presence of mutation. There was a trend to increased OS in HPV+ pts (HR 0.47, 95% CI 0.21–1.07, P=0.068). In paired biopsies, some expression variation was seen for cytoplasm AKT, membrane EGFR, nuclear ERK and pAKT. There was no correlation between basal expression of these markers and RR/CB or PFS. Variations in ratio to baseline of EGFR, pAKT, pERK and MET were qualitatively associated with RR/CB. No statistically significant correlations could be established for PFS, but there were interesting qualitative variations in the levels of expression of some molecules, eg, EGFR, pAKT. Conclusions: No predictive efficacy marker was identified. It cannot be determined if increased OS in HPV+ cases is due to prognostic or predictive effects. Paired biopsies demonstrated that dacomitinib was associated with variation in expression of multiple elements in signalling pathways linked to EGFR. Given the small number of paired biopsies, and large amount of data generated, descriptive study of cases is required. Further data will be presented at the meeting. Clinical trial information: 00768664.

Tu2130 Anti-Mouse IL-6 Receptor Antibody Suppresses Intestinal Carcinogenesis in APCMIN/+ Mice Receiving a High-Fat Diet

9, respectively) were cultured on the rat or human subcutaneous adipose tissue-embedded or -nonembedded gel. Adipose tissue promoted the expression of the growth markers, Ki67 antigen and 24 h-labeling bromodeoxyuridine (BrdU) in the cancer cell types, whereas it inhibited that of the apoptosis marker, cleaved caspase 3. Adipose tissue promoted the basal and superficial expression of the differentiation markers, p63 and involucrin, respectively, within the epithelial layer formed by cancer cell types. Adipose tissue accelerated the invasion of cancer cell types into the gel, together with increased expression of filamin A, laminin-5 and membrane type 1-matrix metalloproteinase (MT1-MMP), and with decreased display of E-cadherin. Adipose tissue promoted the expression of mitogen-activated protein kinase (MAPK: Raf-1, MEK-1 and pERK-1/2) and phosphoinositide 3-kinase-Akt (PI3K-Akt: PTEN, p-4E-BP1 and S6) pathways, and insulin-like growth factor-I receptor (IGF-IR) in the cell types, while it decreased that of the molecularly targeted protein, human epidermal growth factor receptor 2 (HER2). Adipose tissue did not affect the expression of the prostaglandin biosynthetic enzyme cyclooxgenase-2 (COX-2) in the cell types. The COX-2 inhibitor celecoxib did not affect the morphology and invasion of the cell types. Cancer cell types in turn decreased adiponection, leptin and registin production in adipose tissue. The data suggest, first, that adipose tissue may influence the progression of esophageal cancer with the increased growth/invasion and the decreased apoptosis through MAPK, PI3K-Akt and IGF-IR up-regulation in a COX-2-independent way, although adipose tissue seems to induce the differentiation of cancer cells; second, that adipose tissue may adversely affect the HER2targeted therapy; and third, that the cancer cells may affect adipokine production of adipose tissue. Collectively, we conclude that adipose tissue may be involved in the progression of esophageal cancer under adipose tissue-cancer cell interaction.

Tumor Grade and Matrix Metalloproteinase 2 Expression in Stromal Fibroblasts Help to Stratify the High-Risk Group of Patients With Early Breast Cancer Identified on the Basis of St Gallen Recommendations

BackgroundIt is still being discussed if the assessment of basal markers or if adhesion molecules expression contributes additional prognostic information to the classic prognostic factors and hence should be included into standard morphologic reports. Patients and MethodsThe aim of the study was to assess the prognostic significance of: (i) classification recommended by St Gallen experts (ii) tumor grade, expression of (iii) basal markers, (iv) adhesion molecules, and (v) matrix metalloproteinase 2 (MMP-2) in patients with T1-T2 N0M0 chemotherapy-naive ductal breast cancer. ResultsIn 79 patients with tumors characterized by estrogen receptor (ER) and progesterone receptor (PgR) positive, human epidermal growth factor receptor 2 negative (HER2) phenotype and MIB-1 labeling index (MIB-l) LI ≤ 15% (low-risk group) cumulative 17-year breast cancer–specific survival probability was 100% and was significantly higher than in 95 patients from the high-risk group (ER−/PgR−/HER2− or HER2+ or MIB-1 LI > 15%) (72.5%). We found that MMP-2 fibroblast expression indicated 2 subgroups with significantly different survival rates in women with grade 3 tumor (88.9% for MMP-2 positivity and 56.0% for negativity). Cox multivariate analysis revealed that both grade 3 combined with stromal fibroblast MMP-2− and a high-risk group according to St Gallen recommendations are independent negative prognostic factors that influence survival of patients with breast cancer. ConclusionTo the best of our knowledge, we have shown for the first time that MMP-2− in stromal fibroblasts might indicate poor survivors in the group of patients with grade 3 tumors and that the cumulative effect of both above-mentioned parameters might be helpful in selecting the high-risk individuals from the group of patients with luminal B subtype/HER2+/triple negative phenotype identified according to St Gallen recommendations.

Clinical and Molecular Significance of Poly (ADP-Ribose) Polymerase-1 (PARP-1) in Breast Cancer of African Women and its Potential as a Targeted Therapy

Background: The therapeutic effects of Poly (ADP-ribose) polymerase-1 (PARP -1) inhibition are currently studied in a clinical trial that is recruiting African- American (A-A) women with breast cancer (BC). Although, A-A and West African women are likely to share the same ancestry, there are overwhelming evidences, that BC is undoubtedly heterogeneous which might influence results obtained in these Nationalities. Thus, this study aims to investigate PARP-1 expression in a large and annotated series of breast cancer from Nigerian women to determine its clinical and biological significance for the indigenous African women. Methods: PARP-1 protein expression was assessed immunohistochemically in 204 formalin fixed paraffin samples from Nigerian breast cancer women prepared as TMA. Results: PARP-1 was inversely associated with steroid hormone receptors (oestrogen (ER) and progesterone (PR) receptor), the Homologous Recombination marker BRCA1 associated ring domain 1 (BARD1) and p27. Conversely, a positive association was established between PARP-1 and high histologic grade, expression of basal markers (cytokeratins (CK) 5/6 and 14) and epidermal growth factor receptor (EGFR)), DNA damage-repair markers (protein inhibitor of activator signal transducer gamma (PIAS)), the BRCA1 inhibitor (metastasis tumour antigen-1 (MTA1), p53, the proliferation markers (KI-67, Phosphoinositide-3-kinases (PI3KCA)), the triple-negative and basal-like phenotypes. Outcome analysis indicated PARP-1 as a predictor of poor survival independent of tumour size, histological grade and lymph node involvement. Conclusion: These results provide evidence that PARP-1 plays an important role in Nigerian women with breast cancer. It is recommended that indigenous Black women from Africa are included in the ongoing clinical trial of PARP1 inhibitors that is aimed at determining the efficiency of the drug in black BC women outside United States.

Immunohistochemical Profile and Morphology in Triple – Negative Breast Cancers

Triple Negative Breast Cancer (TNBC) is defined by a lack of expression of the steroid hormone receptors (oestrogen and progesterone), and the human epidermal growth factor receptor-2 (HER-2). It is characterized by distinct molecular, histological and clinical features. It is a high risk breast cancer that lacks the benefit of a specific therapy. Our study was aimed at pathologically illustrating triple-negative breast carcinoma and at evaluating the expression of the Epidermal Growth Factor Receptor (EGFR) ,cytokeratin 5/6 (CK 5/6) and Ki-67 among triple-negative breast cancer cases. Further, we aimed to probe whether triple-negative phenotype could be a surrogate marker for the basal phenotype and to correlate the expression of the basal markers (CK 5/6 and EGFR) with the clinico-pathological prognostic parameters. The expression of EGFR, CK 5/6 and Ki-67 were studied by immunohistochemistry (IHC) in 50 triple-negative breast cancer cases. A statistical analysis was implemented by using the SPSS version 20.0. The Chi-square test was conducted to assess the relationship between the immunohistochemical markers and other variables. The Fischer exact test was used when the expected cell counts were less than 5. The women with triple-negative breast cancer were younger, with the adverse pathological characteristics of a high tumour grade, tumour necrosis, frequent nodal metastases and high proliferation. 37 (74%) of the 50 triple-negative breast carcinomas showed the expression of the basal markers (EGFR and /or CK 5/6). We observed a statistically significant association between the basal marker expression and the presence of tumour necrosis. The triple-negative breast cancers in our population harbour adverse pathobiological features and a five marker immunohistochemical panel can be reliably used to define the basal-like cancers. The "Triple-negative" status cannot be used as a surrogate for the "basal marker expression".

Atbf1 Regulates Pubertal Mammary Gland Development Likely by Inhibiting the Pro-Proliferative Function of Estrogen-ER Signaling

ATBF1 is a candidate tumor suppressor that interacts with estrogen receptor (ER) to inhibit the function of estrogen-ER signaling in gene regulation and cell proliferation control in human breast cancer cells. We therefore tested whether Atbf1 and its interaction with ER modulate the development of pubertal mammary gland, where estrogen is the predominant steroid hormone. In an in vitro model of cell differentiation, i.e., MCF10A cells cultured in Matrigel, ATBF1 expression was significantly increased, and knockdown of ATBF1 inhibited acinus formation. During mouse mammary gland development, Atbf1 was expressed at varying levels at different stages, with higher levels during puberty, lower during pregnancy, and the highest during lactation. Knockout of Atbf1 at the onset of puberty enhanced ductal elongation and bifurcation and promoted cell proliferation in both ducts and terminal end buds of pubertal mammary glands. Enhanced cell proliferation primarily occurred in ER-positive cells and was accompanied by increased expression of ER target genes. Furthermore, inactivation of Atbf1 reduced the expression of basal cell markers (CK5, CK14 and CD44) but not luminal cell markers. These findings indicate that Atbf1 plays a role in the development of pubertal mammary gland likely by modulating the function of estrogen-ER signaling in luminal cells and by modulating gene expression in basal cells.

Relationships Between Immunophenotype, Ki-67 Index, Microvascular Density, Ep-CAM/P-cadherin, and MMP-2 Expression in Early-stage Invasive Ductal Breast Cancer

There is still a lack of complete consensus on immunohistochemical surrogate markers for luminal A (LA) and luminal B (LB), HER2, and basal-like subtypes of breast carcinomas and their correlation with cancer cell adhesion and invasion-promoting factors. Therefore, early-stage invasive ductal breast cancer patients (N=209) were recruited to the study and divided into 4 subtypes, on the basis of the expression of the estrogen/progesterone receptor and HER2 (LA: 74.4% of cases; LB: 7.8%; HER2: 5.6%; and triple-negative phenotype: 12.2%). Regardless of the above-mentioned classification, we divided all carcinomas into 2 groups: carcinomas expressing at least 1 basal marker [cytokeratine (CK)5/6, CK5, vimentin, epidermal growth factor receptor, or aberrant CK8/18 expression-membranous or in <10% of cells] versus carcinomas negative for basal markers. Then we studied the relationships between the above subtypes (2 classifications) and (i) the expression of adhesion molecules (Ep-CAM, P-cadherin), (ii) matrix metalloproteinases (MMP)-2, (iii) the proliferation index (MIB-1 LI), and (iv) the microvascular density. We confirmed that triple-negative phenotypes are characterized by basal marker expression, a high tumor grade, and high MIB-1 LI. In this subtype, we found MMP-2 expression in stromal leukocytes less frequently. Both LA carcinomas and carcinomas negative for basal markers were more often negative for epithelial cell adhesion molecule (Ep-CAM) and P-cadherin. Moreover, we noted a higher mean value of microvascular density in CK5/6 and Ep-CAM-immunopositive tumors, carcinomas with aberrant CK8/18 expression, and carcinomas with no or strong expression of MMP-2 in stromal fibroblast-like cells. These results might suggest that mechanisms of stroma remodeling and carcinogenesis (Ep-CAM is the suggested marker of breast progenitors) may differ between breast cancer subtypes.

Cancer stem cell markers are associated with adverse biomarker profiles and molecular subtypes of breast cancer

Cumulative evidence has demonstrated the presence of cancer stem cells (CSC) in breast cancer and its putative role in cancer progression. Nonetheless, the clinical significance of CSC in breast cancer remains elusive. The underlying reasons could be due to the heterogeneity of breast cancer subtypes as well as different markers used to define breast CSC. In this study, three widely used markers (aldehyde dehydrogenase (ALDH)1+ and CD24-CD44+) were used to define two populations of CSC in a large cohort of breast cancers. The expressions of these markers were correlated with different clinicopathological features and the molecular subtypes. ALDH1+ breast cancers were associated with basal-like and HER2-overexpressing subtypes and the characteristics histologic features were related to these two subtypes. On the other hand, CD24-CD44+ breast cancers were associated positively with the presence of extensive in situ component, the absence of lymph node involvement, and basal markers, but negatively with HER2. CD24-CD44+ breast cancers were also positively associated with luminal B cancers. As the expression of CSC markers varied among different molecular subtypes and different clinicopathological features, it appeared that each CSC population could have distinct clinical values in different subgroups of breast cancers. For improved prognostication with CSC, combining the analysis of CSC markers would be required. Within the luminal cancers, CSC appeared to identify cancers with poor outcome. The presence of CSC populations was associated with ER-PR+ cancers and tumors expressing basal markers. Basal marker expression can complement with CSC for improved indicator for poor prognosis in luminal breast cancers. For the first time, the possible contribution of CSC to these aggressive luminal cancers was demonstrated. The association of basal features and CSC in luminal cancers also raised the possibility that luminal cancer cells may acquire basal phenotype and CSC properties together during their progression.

Immunohistochemical characterization of molecular subtypes of invasive breast cancer: a study from North India

In recent years breast cancer has been classified on the basis of its molecular characteristics by gene expression profiling. A similar classification using immunohistochemistry has been identified so that it has a wider application. This study was designed to define the precise prevalence of molecular subtypes of invasive breast carcinoma using immunohistochemistry in patients from north India and to correlate it with known clinical and histological prognostic factors. Based on ER/PR/Her2/neu expression, 100 cases of invasive breast cancer were categorized into: ER+ and ⁄ or PR+ and Her2/neu- (47%), ER+ and⁄or PR+ and Her2/neu+ (15%), ER- and ⁄ or PR- and Her2/neu+ (Her2/neu overexpressing, 21%), ER-, PR- and Her2/neu- (Triple negative, 17%). All cases demonstrated positivity for the luminal Cytokeratins 8/18. In addition, 10% of these tumours showed expression of the basal markers (CK4/14, CK5/6). Among the 17 triple negative cases, eight cases were positive for one of the basal markers and two cases with basal marker expression were Her2/neu overexpressing. The basal markers showed significant correlations only with histological grade and ER negative status. On the basis of hormone receptor, Her-2/neu and cytokeratin expressions, distinct subclasses of breast cancer have been identified which show significant differences in relation to histological grade and ER status. Expression of basal markers is needed to define basal-like breast cancer.