Expression of EGF receptors in canine prostate with proliferative inflammatory atrophy and carcinoma

<div><p>ABSTRACT: Gene expression of ErbB1 and ErbB2, and immunostaining of EGFR (Her1) and Her2 (c-erbB-2) were evaluated in this study to ascertain whether these receptors are involved in the evolution of canine premalignant and malignant prostatic lesions, as proliferative inflammatory atrophy (PIA) and prostatic carcinoma (PC). With regards to the intensity of EGFR immunostaining, there was no difference between normal prostatic tissue and tissues with PIA or PC. In relation to Her2 immunostaining, there were differences between normal prostatic tissue and those with PIA and PC, as also differences between prostates with PIA and PC. There was no correlation between EGFR and Her2 immunostaining. ErbB1 gene product was detected in two normal tissue samples, in one with PIA, and in all samples with PC. ErbB2 mRNA was recorded in two canine samples with PIA, in all with PC, but was not detected in normal prostatic tissue. It was concluded that EGFR and Her2 play roles in canine PIA and PC, suggesting that those receptors may be involved in canine prostatic carcinogenesis.</p></div

An Integrative Genomic and Transcriptomic Analysis Reveals Potential Targets Associated with Cell Proliferation in Uterine Leiomyomas

<div><p>Background</p><p>Uterine Leiomyomas (ULs) are the most common benign tumours affecting women of reproductive age. ULs represent a major problem in public health, as they are the main indication for hysterectomy. Approximately 40–50% of ULs have non-random cytogenetic abnormalities, and half of ULs may have copy number alterations (CNAs). Gene expression microarrays studies have demonstrated that cell proliferation genes act in response to growth factors and steroids. However, only a few genes mapping to CNAs regions were found to be associated with ULs.</p> <p>Methodology</p><p>We applied an integrative analysis using genomic and transcriptomic data to identify the pathways and molecular markers associated with ULs. Fifty-one fresh frozen specimens were evaluated by array CGH (JISTIC) and gene expression microarrays (SAM). The CONEXIC algorithm was applied to integrate the data.</p> <p>Principal Findings</p><p>The integrated analysis identified the top 30 significant genes (<i>P</i><0.01), which comprised genes associated with cancer, whereas the protein-protein interaction analysis indicated a strong association between FANCA and BRCA1. Functional <i>in silico</i> analysis revealed target molecules for drugs involved in cell proliferation, including FGFR1 and IGFBP5. Transcriptional and protein analyses showed that FGFR1 (<i>P</i> = 0.006 and <i>P</i><0.01, respectively) and IGFBP5 (<i>P</i> = 0.0002 and <i>P</i> = 0.006, respectively) were up-regulated in the tumours when compared with the adjacent normal myometrium.</p> <p>Conclusions</p><p>The integrative genomic and transcriptomic approach indicated that <i>FGFR1</i> and <i>IGFBP5</i> amplification, as well as the consequent up-regulation of the protein products, plays an important role in the aetiology of ULs and thus provides data for potential drug therapies development to target genes associated with cellular proliferation in ULs.</p> </div

Data validation.

<p>(A) Boxplot illustrating MM (normal) and ULs (tumour) normalised to obtain relative expression values for all samples evaluated by RT-qPCR. <i>P</i> = paired <i>t</i> test significance. **<i>P</i> = 0.006; ***<i>P</i> = 0.0002; Immunostaining frequency for the (B) FGFR1 and (C) IGFBP5 proteins. The <i>P</i> values (Fisheŕs test) were obtained based on the comparison of the MM and ULs immunostaining results.</p

Seventy-five modulators obtained from integrative analysis.

<p><b>In bold</b>, the top 30 modulators based on the highest scores on CONEXIC; Hg18: Human genome version 18 (Mar 2006 NCBI36); ^*miRNA target prediction; positive (+) and negative (−) signs indicate the gene status with regard to the genomic gains and losses and up- or down-regulated gene expression, respectively. ^£Regions which usually not are involved in chromosomal breakpoints.</p

FGFR1 and IGFBP5 protein expression by immunohistochemistry in uterine leiomyomas.

<p>(A) FGFR1-adjacent normal myometrium showing FGFR1 low level expression (score 1); (B) and (C) FGFR1 cytoplasmic positive expression in uterine leiomyoma tissue (scores 2 and 3/intensity, respectively); (D) adjacent normal myometrium showing IGFBP5 negative expression (score 0); (E) and (F) IGFBP5 cytoplasmic positive expression in uterine leiomyoma tissue (scores 2 and 3/intensity, respectively) (200×).</p

Protein-protein interaction network of 75 modulators.

<p>The 75 modulators were used to query the I2D database to obtain their interacting partners (and also interactions among the modulators). I2D v. 2.0 contained data on 62 modulators, which resulted in a large network of 1,456 proteins and 29,530 interactions. The upward triangles represent up-regulated genes, and the downward triangles represent down-regulated genes. The red and green triangle lines represent genes in amplified and deleted regions, respectively. The biological processes that the modulators are involved are represented by different colours, per the legend, and the triangle size corresponds with the score, i.e., larger triangles depict the highest scores. Direct physical interactions between modulators are represented by blue lines. The remainder of the network is partially transparent to reduce the clutter. The network visualisation and analysis was performed in NAViGaTOR 2.3.</p

Canonical pathway from hepatic fibrosis/hepatic stellate cell activation.

<p>Cell proliferation and extracellular matrix deposition process with high similarity observed in ULs. FGFR1 and IGFBP5 were selected as potential molecular markers in ULs treatment.</p

Hierarchical clustering.

<p>The patients were grouped according to the menstrual cycle phase (proliferative and secretory), number of samples evaluated and diagnosis of multiple or solitary tumours. These results show that the genomic and transcriptomic data were useful to clustering the samples regardless of the clinical features, indicating that could be markers to tumour biology (TMeV v.4.5).</p

EGFR and HER-2 expression by immunohistochemistry (IHC) and their mRNA by <i>in situ</i> hybridization (ISH) in canine mammary carcinomas in mixed tumors and carcinosarcomas.

<p>EGFR and HER-2 expression by immunohistochemistry (IHC) and their mRNA by <i>in situ</i> hybridization (ISH) in canine mammary carcinomas in mixed tumors and carcinosarcomas.</p

Clinicopathological characteristics of canine mammary carcinomas in mixed tumors (CMTs) and carcinosarcomas (CSs).

<p>Clinicopathological characteristics of canine mammary carcinomas in mixed tumors (CMTs) and carcinosarcomas (CSs).</p