PATE,a gene expressed in prostate cancer, normal prostate, and testis, identified by a functional genomic approach

Abstract

To identify target antigens for prostate cancer therapy, we have combined computer-based screening of the human expressed sequence tag database and experimental expression analysis to identify genes that are expressed in normal prostate and prostate cancer but not in essential human tissues. Using this approach, we identified a gene that is expressed specifically in prostate cancer, normal prostate, and testis. The gene has a 1.5-kb transcript that encodes a protein of 14 kDa. We named this gene PATE (expressed in prostate and testis). In situ hybridization shows that PATE mRNA is expressed in the epithelial cells of prostate cancers and in normal prostate. Transfection of the PATE cDNA with a Myc epitope tag into NIH 3T3 cells and subsequent cell fractionation analysis shows that the PATE protein is localized in the membrane fraction of the cell. Analysis of the amino acid sequence of PATE shows that it has structural similarities to a group of proteins known as three-finger toxins, which includes the extracellular domain of the type ␤ transforming growth factor receptor. Restricted expression of PATE makes it a potential candidate for the immunotherapy of prostate cancer.immunotherapy ͉ cancer vaccine ͉ snake toxin ͉ membrane protein P rostate cancer is the most frequently diagnosed noncutaneous cancer and the second leading cause of cancer deaths in men. It has been estimated that about one in five men in the United States will develop prostate cancer during their lifetime, and, to date, there are no curative therapies available for this disease after it has metastasized from its site of origin.

NGEP, a gene encoding a membrane protein detected only in prostate cancer and normal prostate

“…RNA hybridizations with multiple tissue RNA dot-blots and Northern blots were performed as follows: membranes were prehybridized for 2 h in hybridization solution (Hybrisol I, Intergen, Purchase, NY) at 45°C. The DNA probe labeled with 32 P by random primer extension was added to the blot and hybridized for another 16 h. Washing and autoradiography of the blots were performed as described (18).…”

“…Several genes with specific or differential expression in prostate have been identified by this approach but none are membrane proteins (13)(14)(15)(16)(17)(18). In this report, we describe an additional gene that is specifically expressed in prostate cancer and in normal prostate but not in any other tissues.…”

“…The 293T cells were transiently transfected with these plasmids. After transfection, Western blot analysis was done as described (18). Filters were probed with 200 ng͞ml anti-Myc-tag mAb (9E10; Santa Cruz Biotechnology).…”

Targeting lipid metabolism by the lipoprotein lipase inhibitor orlistat results in apoptosis of B-cell chronic lymphocytic leukemia cells

“…[15][16][17] LEP14 and LEP13 18 were predicted in silico to have PLA2 activity, and PATE bears a putative PLA2 motif. 19 Adiponutrin is similar to patatin-like phospholipase domain-containing protein 3 and also shows PLA2 activity. 20 Phospholipase C activity or relation was found in five genes; phospholipase C-b (PLCb1) catalyzes the generation of inositol 1,4,5-trisphosphate and diacylglycerol from phosphatidylinositol 4,5-bisphosphate, a key step in the intracellular transduction of many extracellular signals, 21 whereas PLC-b2 prefers phosphatidyl-inositol 4,5-bisphosphate to phosphatidylinositol as substrate.…”

Cloning, molecular characterization, and expression analysis of Copine 8

“…To achieve this goal, we focused on utilizing the EST database to generate clusters of ESTs that are expressed in normal prostate and/ or prostate cancer. Using this approach, we previously identified genes whose transcripts are specifically expressed in normal prostate and/or prostate cancer [3][4][5][6]. Using a similar approach, we describe here the cloning and characterization of Copine 8, a new member of the gene family that produces the calcium-dependent, phospholipid binding proteins called copines [7][8][9].…”