Expression profiling in ovarian clear cell carcinoma: identification of hepatocyte nuclear factor-1 beta as a molecular marker and a possible molecular target for therapy of ovarian clear cell carcinoma.

Of all of the epithelial ovarian cancers, clear cell carcinoma (CCC) of the ovary has the worst prognosis. We applied the oligonucleotide array technique to identify genes generally involved in CCC. Of the approximately 12,600 genes that were analyzed, 28 were expressed significantly differently between four CCC and seven non-CCC cell lines. Among 16 up-regulated genes in CCC, we further investigated a transcription factor, hepatocyte nuclear factor-1 beta (HNF-1 beta). We validated up-regulation of HNF-1 beta in CCC in terms of both mRNA and protein level using real-time quantitative reverse transcriptase-polymerase chain reaction and immunoblotting. Immunohistochemical analysis of 83 surgically resected ovarian cancers showed that almost all CCC specimens (21 of 22 cases) had nuclear staining for HNF-1 beta, whereas most non-CCC specimens (60 of 61 cases) showed no immunostaining or only focal and faint staining in the nucleus. Furthermore, we investigated the significance of HNF-1 beta expression in CCC using RNA interference. The reduction of HNF-1 beta expression by RNA interference induced apoptotic cell death in ovarian CCC cells, which was confirmed by terminal dUTP nick-end labeling and fluorescence-activated cell-sorting analyses. Our results suggest that HNF-1 beta is not only an excellent CCC-specific molecular marker but also a molecular target for therapy of ovarian CCC.

[1]  R. Berkowitz,et al.  Osteopontin as a potential diagnostic biomarker for ovarian cancer. , 2002, JAMA.

[2]  Mitsuaki Suzuki,et al.  Clinical characteristics of clear cell carcinoma of the ovary , 2000, Cancer.

[3]  T. Tuschl,et al.  Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells , 2001, Nature.

[4]  S. Pinder,et al.  Inhibition of nuclear NAD nucleosidase and poly ADP-ribose polymerase activity from rat liver by nicotinamide and 5'-methyl nicotinamide. , 1971, Biochimica et biophysica acta.

[5]  Hiroyuki Kuroki,et al.  Mutation in hepatocyte nuclear factor–1β gene (TCF2) associated with MODY , 1997, Nature Genetics.

[6]  S. Hirohashi,et al.  Allelotype Analysis of Common Epithelial Ovarian Cancers with Special Reference to Comparison between Clear Cell Adenocarcinoma with Other Histological Types , 2002, Japanese journal of cancer research : Gann.

[7]  Kathleen R. Cho,et al.  Gene expression in ovarian cancer reflects both morphology and biological behavior, distinguishing clear cell from other poor-prognosis ovarian carcinomas. , 2002, Cancer research.

[8]  C. Fisher,et al.  Clear cell epithelial ovarian cancer (mesonephroid): bad prognosis only in early stages. , 1993, Gynecologic oncology.

[9]  M. Pontoglio,et al.  Hepatocyte nuclear factor 1, a transcription factor at the crossroads of glucose homeostasis. , 2000, Journal of the American Society of Nephrology : JASN.

[10]  G. Riggins,et al.  Large-scale serial analysis of gene expression reveals genes differentially expressed in ovarian cancer. , 2000, Cancer research.

[11]  J. Mesirov,et al.  Molecular classification of cancer: class discovery and class prediction by gene expression monitoring. , 1999, Science.

[12]  R. Cortese,et al.  LFB1 and LFB3 homeoproteins are sequentially expressed during kidney development. , 1992, Development.

[13]  Y. Kaneda,et al.  Sp1 decoy transfected to carcinoma cells suppresses the expression of vascular endothelial growth factor, transforming growth factor beta1, and tissue factor and also cell growth and invasion activities. , 2000, Cancer research.

[14]  Ronald W. Davis,et al.  Quantitative Monitoring of Gene Expression Patterns with a Complementary DNA Microarray , 1995, Science.

[15]  Judy Lieberman,et al.  RNA interference targeting Fas protects mice from fulminant hepatitis , 2003, Nature Medicine.

[16]  Michael T. McManus,et al.  A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells and transgenic mice by RNA interference , 2003, Nature Genetics.

[17]  M. Piver.  Ovarian carcinoma: A decade of progress , 1984, Cancer.

[18]  M. Yaniv,et al.  vHNF1 is a homeoprotein that activates transcription and forms heterodimers with HNF1. , 1991, The EMBO journal.

[19]  A. Fire,et al.  Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans , 1998, Nature.

[20]  T. Okai,et al.  p53 mutations and overexpression affect prognosis of ovarian endometrioid cancer but not clear cell cancer. , 2003, Gynecologic oncology.

[21]  T Tanaka,et al.  Identification by cDNA microarray of genes involved in ovarian carcinogenesis. , 2000, Cancer research.

[22]  S. Mundlos,et al.  PAX8, a human paired box gene: isolation and expression in developing thyroid, kidney and Wilms' tumors. , 1992, Development.

[23]  L. Hood,et al.  Monitoring gene expression profile changes in ovarian carcinomas using cDNA microarray. , 1999, Gene.

[24]  R. Carthew,et al.  Use of dsRNA-Mediated Genetic Interference to Demonstrate that frizzled and frizzled 2 Act in the Wingless Pathway , 1998, Cell.

[25]  T. Nikaido,et al.  Clear cell carcinoma has an expression pattern of cell cycle regulatory molecules that is unique among ovarian adenocarcinomas , 1999, Cancer.

[26]  S. Dhanasekaran,et al.  The polycomb group protein EZH2 is involved in progression of prostate cancer , 2002, Nature.

[27]  X. Wang,et al.  Predicting hepatitis B virus–positive metastatic hepatocellular carcinomas using gene expression profiling and supervised machine learning , 2003, Nature Medicine.

[28]  P. Gruss,et al.  The oncogenic potential of Pax genes. , 1993, The EMBO journal.

[29]  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.

[30]  R. Cowan,et al.  A potential role of heat shock proteins and nicotinamide N‐methyl transferase in predicting response to radiation in bladder cancer , 2002, International journal of cancer.

[31]  Quynh-Thu Le,et al.  Identification of osteopontin as a prognostic plasma marker for head and neck squamous cell carcinomas. , 2003, Clinical cancer research : an official journal of the American Association for Cancer Research.

[32]  K. Ariyoshi,et al.  Ovarian endometriosis associated with ovarian carcinoma: a clinicopathological and immunohistochemical study. , 2000, Gynecologic oncology.

[33]  D R Schwartz,et al.  Coordinately up-regulated genes in ovarian cancer. , 2001, Cancer research.

[34]  M. Terris,et al.  Gene expression patterns in renal cell carcinoma assessed by complementary DNA microarray. , 2003, The American journal of pathology.

[35]  R. Lauro,et al.  Pax-8, a paired domain-containing protein, binds to a sequence overlapping the recognition site of a homeodomain and activates transcription from two thyroid-specific promoters , 1992, Molecular and cellular biology.

[36]  J. Cain,et al.  Clear cell carcinoma of the ovary: a distinct histologic type with poor prognosis and resistance to platinum-based chemotherapy in stage III disease. , 1996, Gynecologic oncology.

[37]  M. Wells,et al.  Precursor lesions of ovarian epithelial malignancy , 2001, Histopathology.

[38]  K. Kinzler,et al.  Serial Analysis of Gene Expression , 1995, Science.

[39]  Y. Hayashi,et al.  Expression of HNF‐1α and HNF‐1β in various histological differentiations of hepatocellular carcinoma , 1998 .

[40]  I. Tannock,et al.  Paclitaxel plus carboplatin versus standard chemotherapy with either single-agent carboplatin or cyclophosphamide, doxorubicin, and cisplatin in women with ovarian cancer: the ICON3 randomised trial , 2002, The Lancet.

[41]  J. P. Geisler,et al.  Clear cell carcinoma of the ovary: poor prognosis compared to serous carcinoma. , 1998, European journal of gynaecological oncology.

[42]  P. Clement,et al.  Tumors of the Ovary, Maldeveloped Gonads, Fallopian Tube, and Broad Ligament: Atlas of Tumor Pathology , 1999 .

[43]  L. Sobin,et al.  Histologic typing of ovarian tumors. , 1987, Archives of pathology & laboratory medicine.

[44]  M. Yaniv,et al.  More potent transcriptional activators or a transdominant inhibitor of the HNF1 homeoprotein family are generated by alternative RNA processing. , 1993, The EMBO journal.

[45]  G. Crabtree,et al.  Purified hepatocyte nuclear factor 1 interacts with a family of hepatocyte-specific promoters. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[46]  M. Byrne,et al.  Dominant-negative Suppression of HNF-1α Results in Mitochondrial Dysfunction, INS-1 Cell Apoptosis, and Increased Sensitivity to Ceramide-, but Not to High Glucose-induced Cell Death* , 2002, The Journal of Biological Chemistry.

[47]  M. Blumenfeld,et al.  A liver-specific factor essential for albumin transcription differs between differentiated and dedifferentiated rat hepatoma cells. , 1988, Genes & development.