Immunohistochemical expression analysis of BLCAP in FFPE breast tissue samples.

<p>(A) No immunostaining was observed in a normal breast tissue section reacted with BLCAP antibody preincubated with immunizing peptide. (B) Immunohistochemical staining of BLCAP protein in a normal breast tissue sample demonstrated the presence of the BLCAP antigen in luminal epithelial cells with weak cytoplasmic expression (black arrow). Marked nuclear expression was also observed occasionally (red arrow). Yellow arrow points to a vessel with moderate immunoreactivity for BLCAP. (C) In a few cases IHC analysis of tumor samples showed that BLCAP was expressed in tumor cells with weak cytoplasmic expression (red arrow). (D) Most cases showed moderate to strong cytoplasmic expression with no detectable nuclear presence (red arrow) but in some cases (E) we could observe strong nuclear expression of BLCAP (red arrow). (F) In a few cases, samples were heterogenous with some cells showing distinct perinuclear immunoreactivity for BLCAP (red arrow). (G) Malignant cells showed stronger immunoreactivity (red arrows) than adjacent normal-looking ducts (black arrow), demonstrating up-regulation of this protein in tumor cells. Yellow arrows point to vessels with strong immunoreactivity for BLCAP. (H) We also observed up-regulation of BLCAP in early lesions where lobular carcinoma in situ cells showed overexpression of this protein (grey arrows) in relation to normal adjacent areas (black arrow), and at levels comparable to invasive carcinoma cells (red arrow).</p

2D PAGE and 2D Western blot analysis of BLCAP protein spot patterns.

<p>(A) COS-1 cells transfected with pZeoSV2 empty vector and labeled with ³⁵S-methionine. (B) COS-1 cells transfected with pZeoSV2– BLCAP overexpressing construct and labeled with ³⁵S-methionine. Radioactive metabolic labeling (³⁵S-methionine) of COS-1 cells was used to ensure the highest detection sensitivity. (C) 2D Western blot of COS-1 cells transfected with pZeoSV2– BLCAP construct detected with anti-BLCAP antibody (10 sec film exposure). (D) 2D gel of proteins from breast tumor 63 stained with silver. (E) 2D Western blot of protein lysate from breast tumor 63 (see D) reacted with anti-BLCAP antibody (1 min film exposure). The positions of the BLCAP protein in the 2D-PAGE gels and corresponding 2D Western blots, are indicated by black arrows. The positions of several reference proteins are indicated by red arrows: ACTB – beta actin; ENO1 -alpha enolase 1; CANX – calnexin; PDI - Protein disulfide-isomerase; TUBA1A - tubulin alpha-1A chain; YWHAZ - 14-3-3 protein zeta/delta. The identity of all reference spots were confirmed by MS analysis.</p

Clinicopathological correlation of BLCAP expression in 2,197 sample TMA.

*<p>Kruskal-Wallis one way analysis of variance on ranks.</p>**<p>Mann-Whitney rank-sum test.</p

Differential expression of BLCAP in tumor samples.

<p>The DCTB 123 patient set, including normal, tumor and lymph node metastasis samples (A), or a subset of matched 62 normal and tumor samples (B) were analyzed by quantitative IHC. Mean intensity scores for each group are indicated by red lines.</p

Clinical characteristics of DCTB tissue specimens.

<p>Clinical characteristics of DCTB tissue specimens.</p

Expression analysis of BLCAP by quantitative IHC of normal specimens (blue bars) and corresponding tumor samples (red bars) of 62 matched cases from the DCTB dataset.

<p>Illustrative IHC images are shown for normal and tumor samples with weak immunoreactivity (N11 and T76, respectively), and for normal and tumor samples showing substantial immunoreactivity for BLCAP (N14 and T63, respectively). Magnification 20X.</p

<i>FGFR1</i> Amplification Is Often Homogeneous and Strongly Linked to the Squamous Cell Carcinoma Subtype in Esophageal Carcinoma

<div><p>Background and Aims</p><p>Amplification of the <i>fibroblast growth factor receptor 1</i> (<i>FGFR1</i>) is believed to predict response to multi-kinase inhibitors targeting <i>FGFR1</i>. Esophageal cancer is an aggressive disease, for which novel targeted therapies are highly warranted.</p><p>Methods</p><p>This study was designed to investigate the prevalence and clinical significance of <i>FGFR1</i> amplification in a tissue microarray containing 346 adenocarcinomas and 254 squamous cell carcinomas of the esophagus, using dual-labeling fluorescence <i>in situ</i> hybridization (FISH) analysis.</p><p>Results</p><p><i>FGFR1</i> amplification, defined as a ratio of <i>FGFR1</i>:centromere 8 copy numbers ≥ 2.0, was more frequently seen in squamous cell carcinoma (8.9% of 202 interpretable cases) than in adenocarcinoma (1.6% of 308; p<0.0001). There was no association between <i>FGFR1</i> amplification and tumor phenotype or clinical outcome. To study potential heterogeneity of <i>FGFR1</i> amplification, all available tumor blocks from 23 <i>FGFR1</i> amplified tumors were analyzed on conventional large sections. This analysis revealed complete homogeneity of <i>FGFR1</i> amplification in 20 (86.9%) primary tumors and in all available lymph node metastases. Remarkably, <i>FGFR1</i> amplification was also seen in dysplasia adjacent to tumor in 6 of 9 patients with <i>FGFR1</i> amplified primary cancers.</p><p>Conclusions</p><p>In conclusion, <i>FGFR1</i> amplification occurs in a relevant subgroup of carcinomas of the esophagus and may play a particular role for development of squamous cell cancers. The high homogeneity of <i>FGFR1</i> amplification suggests that patients with <i>FGFR1</i> amplified esophageal cancers may particularly benefit from anti-<i>FGFR1</i> therapies and prompt for clinical studies in this tumor type.</p></div

Supplementary Table S2 from Integrative Genomic Analysis of Small-Cell Lung Carcinoma Reveals Correlates of Sensitivity to Bcl-2 Antagonists and Uncovers Novel Chromosomal Gains

Supplementary Table S2 from Integrative Genomic Analysis of Small-Cell Lung Carcinoma Reveals Correlates of Sensitivity to Bcl-2 Antagonists and Uncovers Novel Chromosomal Gain

Proteomic Profiling of Mammary Carcinomas Identifies C7orf24, a γ-Glutamyl Cyclotransferase, as a Potential Cancer Biomarker

Breast cancer is the leading cause of cancer deaths in women today and is the most common cancer (excluding skin cancers) among women in the Western world. Although cancers detected by screening mammography are significantly smaller than nonscreening ones, noninvasive biomarkers for detection of breast cancer as early as possible are an urgent need as the risk of recurrence and subsequent death is closely related to the stage of the disease at the time of primary surgery. A set of 123 primary breast tumors and matched normal tissue was analyzed by two-dimensional (2D) gel electrophoresis, and a novel protein, C7orf24, was identified as being upregulated in cancer cells. Protein expression levels of C7orf24 were evaluated by immunohistochemical assays to qualify deregulation of this protein. Analysis of C7orf24 expression showed up-regulation in 36.4 and 23.4% of cases present in the discovery sample set (123 samples) and in an independent large TMA validation data set (2197 samples) of clinically annotated breast cancer specimens, respectively. Survival analysis showed that C7orf24 overexpression defines a subgroup of breast tumors with poor clinical outcome. Up-regulation of C7orf24 was also found in other cancer types. Four of these were investigated in greater detail, and we found that a proportion of tumors (58% in cervical, 38% in lung, 72% in colon, and 46% in breast cancer) expressed C7orf24 at levels exceeding those seen in normal samples. The observed overexpression of this protein in different types of cancer suggests deregulation of C7orf24 to be a general event in epithelial carcinogenesis, indicating that this protein may play an important role in cancer cell biology and thus constitute a novel therapeutic target. Furthermore, as C7orf24 is externalized to the tissue extracellular fluid and can be detected in serum, this protein also represents a potential serological marker

Esophageal Carcinoma—Array.

<p>Esophageal Carcinoma—Array.</p