Identification of differentially expressed proteins in ovarian cancer using high-density protein microarrays

Ovarian cancer is a leading cause of deaths, yet many aspects of the biology of the disease and a routine means of its detection are lacking. We have used protein microarrays and autoantibodies from cancer patients to identify proteins that are aberrantly expressed in ovarian tissue. Sera from 30 cancer patients and 30 healthy individuals were used to probe microarrays containing 5,005 human proteins. Ninety-four antigens were identified that exhibited enhanced reactivity from sera in cancer patients relative to control sera. The differential reactivity of four antigens was tested by using immunoblot analysis and tissue microarrays. Lamin A/C, SSRP1, and RALBP1 were found to exhibit increased expression in the cancer tissue relative to controls. The combined signals from multiple antigens proved to be a robust test to identify cancerous ovarian tissue. These antigens were also reactive with tissue from other types of cancer and thus are not specific to ovarian cancer. Overall our studies identified candidate tissue marker proteins for ovarian cancer and demonstrate that protein microarrays provide a powerful approach to identify proteins aberrantly expressed in disease states.

[1]  Y. Awasthi,et al.  Depletion of RLIP76 sensitizes lung cancer cells to doxorubicin. , 2005, Biochemical pharmacology.

[2]  P. Brown,et al.  Autoantigen microarrays for multiplex characterization of autoantibody responses , 2002, Nature Medicine.

[3]  Donald L. Weaver,et al.  Three-dimensional reconstruction by confocal laser scanning microscopy in routine pathologic specimens of benign and malignant lesions of the human breast , 1997, Histochemistry and Cell Biology.

[4]  D. Reinberg,et al.  Interaction of FACT, SSRP1, and the High Mobility Group (HMG) Domain of SSRP1 with DNA Damaged by the Anticancer Drug Cisplatin* , 2001, The Journal of Biological Chemistry.

[5]  Mark Gerstein,et al.  ProCAT: a data analysis approach for protein microarrays , 2006, Genome Biology.

[6]  Donald R Schwartz,et al.  Comprehensive proteome analysis of ovarian cancers using liquid phase separation, mass mapping and tandem mass spectrometry: A strategy for identification of candidate cancer biomarkers , 2004, Proteomics.

[7]  Gerard Tromp,et al.  Diagnostic markers of ovarian cancer by high-throughput antigen cloning and detection on arrays. , 2006, Cancer research.

[8]  Bharti Odhav,et al.  Immune responses in cancer. , 2003, Pharmacology & therapeutics.

[9]  A. Jemal,et al.  Cancer Statistics, 2006 , 2006, CA: a cancer journal for clinicians.

[10]  G. Auer,et al.  Phenotypic analysis of ovarian carcinoma: Polypeptide expression in benign, borderline and malignant tumors , 1997, International journal of cancer.

[11]  M. Schummer,et al.  Serologic analysis of ovarian tumor antigens reveals a bias toward antigens encoded on 17q , 2003, International journal of cancer.

[12]  N. Maraldi,et al.  A pathogenic mechanism leading to partial lipodistrophy and prospects for pharmacological treatment of insulin resistance syndrome. , 2007, Acta bio-medica : Atenei Parmensis.

[13]  Hongyu Zhao,et al.  Serum protein markers for early detection of ovarian cancer. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[14]  R. Bast,et al.  A radioimmunoassay using a monoclonal antibody to monitor the course of epithelial ovarian cancer. , 1983 .

[15]  M. Fraga,et al.  Inactivation of the lamin A/C gene by CpG island promoter hypermethylation in hematologic malignancies, and its association with poor survival in nodal diffuse large B-cell lymphoma. , 2005, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[16]  Cecelia E Schmalbach,et al.  Molecular profiling of the immune response in colon cancer using protein microarrays: Occurrence of autoantibodies to ubiquitin C‐terminal hydrolase L3 , 2003, Proteomics.

[17]  T. Soussi,et al.  p53 Antibodies in the sera of patients with various types of cancer: a review. , 2000, Cancer research.

[18]  Steven J Skates,et al.  Preoperative sensitivity and specificity for early-stage ovarian cancer when combining cancer antigen CA-125II, CA 15-3, CA 72-4, and macrophage colony-stimulating factor using mixtures of multivariate normal distributions. , 2004, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[19]  N. Lemoine,et al.  Intrinsic Chemoresistance to Gemcitabine Is Associated with Decreased Expression of BNIP3 in Pancreatic Cancer , 2005, Clinical Cancer Research.

[20]  Francis S. Collins,et al.  A lamin A protein isoform overexpressed in Hutchinson–Gilford progeria syndrome interferes with mitosis in progeria and normal cells , 2007, Proceedings of the National Academy of Sciences.

[21]  Joshua LaBaer,et al.  The sentinel within: exploiting the immune system for cancer biomarkers. , 2005, Journal of proteome research.

[22]  D. Lockhart,et al.  Analysis of gene expression profiles in normal and neoplastic ovarian tissue samples identifies candidate molecular markers of epithelial ovarian cancer. , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[23]  D. Landsittel,et al.  Circulating IL-8 and anti-IL-8 autoantibody in patients with ovarian cancer. , 2006, Gynecologic oncology.

[24]  Y. Awasthi,et al.  Novel function of human RLIP76: ATP-dependent transport of glutathione conjugates and doxorubicin. , 2000, Biochemistry.

[25]  G. Gatta,et al.  Survival of European women with gynaecological tumours, during the period 1978-1989. EUROCARE Working Group. , 1998, European journal of cancer.

[26]  B. Morris,et al.  ZNF265—a novel spliceosomal protein able to induce alternative splicing , 2001, The Journal of cell biology.

[27]  M. Kirschner,et al.  Homologies in both primary and secondary structure between nuclear envelope and intermediate filament proteins , 1986, Nature.