A Fortran program blue called EMMIX-WIRE (EM-based MIXture analysis WIth Random Effects) is available on request from the corresponding author.

Selenium binding protein 1 (SELENBP1) was identified to be the most significantly down-regulated protein in ovarian cancer cells by a membrane proteome profiling analysis. SELENBP1 expression levels in 4 normal ovaries, 8 benign ovarian tumors, 12 borderline ovarian tumors and 141 invasive ovarian cancers were analyzed with immunohistochemical assay. SELENBP1 expression was reduced in 87% cases of invasive ovarian cancer (122/141) and was significantly reduced in borderline tumors and invasive cancers (p < 0.001). Cox multivariate analysis within the 141 invasive cancer tissues showed that SELENBP1 expression score was a potential prognostic indicator for unfavorable prognosis of ovarian cancer (hazard ratio [HR], 2.18; 95% CI 5 1.22-3.90; p 5 0.009). Selenium can disrupt the androgen pathway, which has been implicated in modulating SELENBP1 expression. We investigated the effects of selenium and androgen on normal human ovarian surface epithelial (HOSE) cells and cancer cells. Interestingly, SELENBP1 mRNA and protein levels were reduced by androgen and elevated by selenium treatment in the normal HOSE cells, whereas reversed responses were observed in the ovarian cancer cell lines. These results suggest that changes of SELENBP1 expression in malignant ovarian cancer are an indicator of aberration of selenium/androgen pathways and may reveal prognostic information of ovarian cancer. ' 2005 Wiley-Liss, Inc.Key words: ovarian cancer; membrane proteome profiling; 2-D PAGE; selenium binding protein 1; prognostic marker; androgen; methylselenocysteine Despite advances in cancer therapeutic agents in recent years, approximately 14,000 women still die of ovarian cancer each year in the United States, making it the most lethal of the gynecological malignancies. 1 Currently, surgical debulking followed by chemotherapy is the major treatment approach for ovarian cancer. Most cases of ovarian cancer are diagnosed at advanced stages when the prognosis for 5-year survival is poor. 2 Developing new diagnostic technique and improving current therapeutic strategy are the main directions to fight this morbid disease.Multiple genomic and proteomic approaches have been applied to identify disease-associated biomarkers for diagnosis and disease management. Applying cDNA and oligo microarray technology have enabled scientists to look into the global differences in gene expression between normal and cancer cells. 3 For proteomic approaches, two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) followed by protein identification using mass spectrometry has been the primary technique for biomarker discovery. 4,5 Membrane-associated proteins have been suggested to be a potential resource for biomarker screening. 6 In our pilot study of the use of 2-D PAGE to profile membrane-associated proteins of normal ovarian surface epithelial cells and ovarian cancer cell lines, selenium binding protein 1 (SELENBP1) was identified as the most significantly down-regulated protein in the cancer cells.The human selenium binding protein gene (...

BACKGROUND Endometriosis is frequently associated with and thought of having propensity to develop into ovarian clear cell carcinoma (OCCC), although the molecular transformation mechanism is not completely understood. METHODS We employed immunohistochemical (IHC) staining for marker expression along the potential progression continuum. Expression profiling of microdissected endometriotic and OCCC cells from patient-matched formalin-fixed, paraffin-embedded samples was performed to explore the carcinogenic pathways. Function of novel biomarkers was confirmed by knockdown experiments. RESULTS PTEN was significantly lost in both endometriosis and invasive tumor tissues, while estrogen receptor (ER) expression was lost in OCCC relative to endometriosis. XRCC5, PTCH2, eEF1A2, and PPP1R14B were significantly overexpressed in OCCC and associated endometriosis, but not in benign endometriosis (p≤0.004). Knockdown experiments with XRCC5 and PTCH2 in a clear cell cancer cell line resulted in significant growth inhibition. There was also significant silencing of a panel of target genes with histone H3 lysine 27 trimethylation, a signature of polycomb chromatin-remodeling complex in OCCC. IHC confirmed the loss of expression of one such polycomb target gene, the serous ovarian cancer lineage marker WT1 in OCCC, while endometriotic tissues showed significant co-expression of WT1 and ER. CONCLUSIONS Loss of PTEN expression is proposed as an early and permissive event in endometriosis development, while the loss of ER and polycomb-mediated transcriptional reprogramming for pluripotency may play an important role in the ultimate transformation process. Our study provides new evidence to redefine the pathogenic program for lineage-specific transformation of endometriosis to OCCC.

Epithelial ovarian cancer is the leading cause of death among gynaecologic cancers in Western countries. Our studies have shown that casein kinase I‐epsilon (CKIε), a Wnt pathway protein, is significantly overexpressed in ovarian cancer tissues and is associated with poor survival. Ectopic expression of CKIε in normal human ovarian surface epithelial cells and inhibition of CKIε in ovarian cancer cells and in xenografts demonstrated the importance of CKIε in regulating cell proliferation and migration. Interestingly, CKIε function did not seem to involve β‐catenin activity. Instead, CKIε was found to interact with several mitochondrial proteins including adenine nucleotide translocase 2 (ANT2). Inhibition of CKIε in ovarian cancer cells resulted in suppression of ANT2, downregulation of cellular ATP and the resulting cancer cells were more susceptible to chemotherapy. Our studies indicate that, in the context of ovarian cancer, the interaction between CKIε and ANT2 mediates pathogenic signalling that is distinct from the canonical Wnt/β‐catenin pathway and is essential for cell proliferation and is clinically associated with poor survival.

Glycoprotein nmb (GPNMB) promotes breast tumor growth and metastasis and its expression in tumor epithelium correlates with poor prognosis in breast cancer patients. Despite its biological and clinical significance, little is known regarding the molecular mechanisms engaged by GPNMB. Herein, we show that GPNMB engages distinct functional domains and mechanisms to promote primary tumor growth and metastasis. We demonstrate that neuropilin-1 (NRP-1) expression is increased in breast cancer cells that overexpress GPNMB. Interestingly, the GPNMB-driven increase in NRP-1 expression potentiated vascular endothelial growth factor signaling in breast cancer cells and was required for the growth, but not metastasis, of these cells in vivo. Interrogation of RNAseq data sets revealed a positive correlation between GPNMB and NRP-1 levels in human breast tumors. Furthermore, we ascribe pro-growth and pro-metastatic functions of GPNMB to its ability to bind α5β1 integrin and increase downstream signaling in breast cancer cells. We show that GPNMB enhances breast cancer cell adhesion to fibronectin, increases α5β1 expression and associates with this receptor through its RGD motif. GPNMB recruitment into integrin complexes activates Src and Fak signaling pathways in an RGD-dependent manner. Importantly, both the RGD motif and cytoplasmic tail of GPNMB are required to promote primary mammary tumor growth; however, only mutation of the RGD motif impaired the formation of lung metastases. Together, these findings identify novel and distinct molecular mediators of GPNMB-induced breast cancer growth and metastasis.

Identification of comorbidity patterns of health conditions is critical for evidence-based practice to improve the prevention, treatment and health care of relevant diseases. Existing approaches focus mainly on either using descriptive measures of comorbidity in terms of the prevalence of coexisting conditions, or addressing the prevalence of comorbidity based on a particular disease (e.g. psychosis) or a specific population (e.g. hospital patients). As coincidental comorbidity by chance increases with the prevalence rates of the conditions, which in turn depend heavily on the population under study, research findings on comorbidity patterns using those approaches may provide unreliable results. In this paper, we propose an asymmetric version of Somers' D statistic to provide a quantitative measure of comorbidity that accounts for co-occurrence of conditions by chance, and develop a unified clustering algorithm to identify comorbidity patterns with adjustment for multiple testing and control for the false discovery rate. We assess the applicability of the proposed comorbidity measure and investigate the performance of the proposed procedure for the adjustment of multiple testing by conducting a comparative study and a sensitivity analysis, respectively. The proposed method is illustrated using a national survey data set of mental health and wellbeing and a national health survey data set in Australia.

Kallikrein 6 (hK6, also known as protease M/zyme/neurosin) is a member of the human kallikrein gene family. We have previously cloned the cDNA for this gene by differential display and shown the overexpression of the mRNA in breast and ovarian primary tumour tissues and cell lines. To thoroughly characterise the expression of this kallikrein in ovarian cancer, we have developed a novel monoclonal antibody specific to hK6 and employed it in immunohistochemistry with a wide range of ovarian tumour samples. The expression was found elevated in 67 of 80 cases of ovarian tumour samples and there was a significant difference in the expression levels between normal and benign ovarian tissues and the borderline and invasive tumours (Po0.001). There was no difference of expression level between different subtypes of tumours. More significantly, high level of kallikrein 6 expression was found in many early-stage and low-grade tumours, and elevated hK6 proteins were found in benign epithelia coexisting with borderline and invasive tissues, suggesting that overexpression of hK6 is an early phenomenon in the development of ovarian cancer. Quantitative real-time reverse transcription -polymerase chain reactions also showed elevated kallikrein 6 mRNA expression in ovarian tumours. Genomic Southern analysis of 19 ovarian tumour samples suggested that gene amplification is one mechanism for the overexpression of hK6 in ovarian cancer.

Transforming growth factor (TGF)-b signaling is a potent modulator of the invasive and metastatic behavior of breast cancer cells. Indeed, breast tumor responsiveness to TGF-b is important for the development of osteolytic bone metastases. However, the specific TGF-b isoforms that promote breast cancer outgrowth in bone is unknown. We demonstrate that expression of a TGF-b ligand trap, which neutralizes TGF-b1 and TGF-b3, in MDA-MB-231 breast cancer cells diminished their outgrowth in bone and reduced the severity of osteolytic lesion formation when compared with controls. We further show that a reduction or loss of TGF-b1 expression within the bone microenvironment of TGF-b1 þ /À and TGF-b1À/À mice significantly reduced the incidence of breast tumor outgrowth compared with wild-type animals. Interestingly, those tumors capable of growing within the tibiae of TGF-b1-deficient mice had upregulated expression of all three TGF-b isoforms. Finally, breast cancer cells expressing the TGF-b ligand trap showed a pronounced reduction in their ability to form osteolytic lesions when injected into the tibiae of TGF-b1 þ /À mice. Thus, our studies show that both host-and tumor-derived TGF-b expression plays a critical role during the establishment and outgrowth of breast cancer cells in bone.