Identification of differentially expressed genes in human prostate cancer using subtraction and microarray.

We have identified human prostate cancer- and tissue-specific genes using cDNA library subtraction in conjunction with high throughput microarray screening. Subtracted cDNA libraries of prostate tumors and normal prostate tissue were generated. Characterization of subtracted libraries showed enrichment of both cancer- and tissue-specific genes. Highly redundant clones were eliminated by colony hybridization. The remaining clones were selected for microarray to determine gene expression levels in a variety of tumor and normal tissues. Clones showing overexpression in prostate tumors and/or normal prostate tissues were selected and sequenced. Here we report the identification of two genes, P503S and P504S, from subtracted libraries and a third gene, P510S, by subtraction followed by microarray screening. Their expression profiles were further confirmed by Northern blot, real-time PCR (TaqMan), and immunohistochemistry to be overexpressed in prostate tissues and/or prostate tumors. Full-length cDNA sequences were cloned, and their subcellular locations were predicted by a bioinformatic algorithm, PSORT, to be plasma membrane proteins. The genes identified through these approaches are potential candidates for cancer diagnosis and therapy.

[1]  A Catalog of Cancer Genes at the Click of a Mouse , 1997, Science.

[2]  P. Chomez,et al.  A gene encoding an antigen recognized by cytolytic T lymphocytes on a human melanoma. , 1991, Science.

[3]  A. Miyajima,et al.  Characterization of cell phenotype by a novel cDNA library subtraction system: expression of CD8 alpha in a mast cell-derived interleukin-4-dependent cell line. , 1994, Blood.

[4]  M. Schwab,et al.  Association Between the Rat Homologue of CO-029, a Metastasis-associated Tetraspanin Molecule and Consumption Coagulopathy , 1998, The Journal of cell biology.

[5]  J. J. Edwards,et al.  Proteins of human urine. II. Identification by two-dimensional electrophoresis of a new candidate marker for prostatic cancer. , 1982, Clinical chemistry.

[6]  A. Pardee,et al.  Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction. , 1992, Science.

[7]  P. Schellhammer,et al.  A novel prostate carcinoma‐associated glycoprotein complex (PAC) recognized by monoclonal antibody turp‐27 , 1991, International journal of cancer.

[8]  K. Honn,et al.  12‐Lipoxygenase, 12(S)‐HETE, and Cancer Metastasis a , 1994, Annals of the New York Academy of Sciences.

[9]  M. Pfreundschuh,et al.  Identification of a meiosis-specific protein as a member of the class of cancer/testis antigens. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[10]  G. Murphy,et al.  Monoclonal antibodies to a new antigenic marker in epithelial prostatic cells and serum of prostatic cancer patients. , 1987, Anticancer research.

[11]  I. Pastan,et al.  Discovery of three genes specifically expressed in human prostate by expressed sequence tag database analysis. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[12]  W. Fair,et al.  Molecular cloning of a complementary DNA encoding a prostate-specific membrane antigen. , 1993, Cancer research.

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

[14]  Ji Huang,et al.  [Serial analysis of gene expression]. , 2002, Yi chuan = Hereditas.

[15]  D. Robinson,et al.  Molecular characterization of the epitope in prostate and breast tumor-associated PR92 antigen. , 1989, Cancer research.

[16]  F C Lowe,et al.  Prostatic acid phosphatase in 1993. Its limited clinical utility. , 1993, The Urologic clinics of North America.

[17]  P. Guinan,et al.  An evaluation of prostate specific antigen in prostatic cancer. , 1987, The Journal of urology.

[18]  G Geisslinger,et al.  Molecular cloning and expression of a 2-arylpropionyl-coenzyme A epimerase: a key enzyme in the inversion metabolism of ibuprofen. , 1997, Molecular pharmacology.

[19]  R H Hruban,et al.  Gene expression profiles in normal and cancer cells. , 1997, Science.

[20]  T. Teni,et al.  Serum and urinary prostatic inhibin-like peptide in benign prostatic hyperplasia and carcinoma of prostate. , 1988, Cancer letters.

[21]  I. Romslo.  Prostatic acid phosphatase. , 1971, British medical journal.

[22]  P. Schellhammer,et al.  Monoclonal antibody PD41 recognizes an antigen restricted to prostate adenocarcinomas. , 1991, Cancer research.

[23]  T. Hasan,et al.  p53 expression and clinical outcome in prostate cancer. , 1993, British journal of urology.

[24]  J. Schrader,et al.  A novel strategy for generating monoclonal antibodies from single, isolated lymphocytes producing antibodies of defined specificities. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[25]  J. Oesterling,et al.  Correlation of clinical stage, serum prostatic acid phosphatase and preoperative Gleason grade with final pathological stage in 275 patients with clinically localized adenocarcinoma of the prostate. , 1987, The Journal of urology.