Identification of the human prostatic carcinoma oncogene PTI-1 by rapid expression cloning and differential RNA display

Proc Natl Acad Sci U S A. 1995 Jul 18;92(15):6778-82. doi: 10.1073/pnas.92.15.6778.

Abstract

Elucidating the relevant genomic changes mediating development and evolution of prostate cancer is paramount for effective diagnosis and therapy. A putative dominant-acting nude mouse prostatic carcinoma tumor-inducing gene, PTI-1, has been cloned that is expressed in patient-derived human prostatic carcinomas but not in benign prostatic hypertrophy or normal prostate tissue. PTI-1 was detected by cotransfecting human prostate carcinoma DNA into CREF-Trans 6 cells, inducing tumors in nude mice, and isolating genes displaying increased expression in tumor-derived cells by using differential RNA display (DD). Screening a human prostatic carcinoma (LNCaP) cDNA library with a 214-bp DNA fragment found by DD permitted the cloning of a full-length 2.0-kb PTI-1 cDNA. Sequence analysis indicates that PTI-1 is a gene containing a 630-bp 5' sequence and a 3' sequence homologous to a truncated and mutated form of human elongation factor 1 alpha. In vitro translation demonstrates that the PTI-1 cDNA encodes a predominant approximately 46-kDa protein. Probing Northern blots with a DNA fragment corresponding to the 5' region of PTI-1 identifies multiple PTI-1 transcripts in RNAs from human carcinoma cell lines derived from the prostate, lung, breast, and colon. In contrast, PTI-1 RNA is not detected in human melanoma, neuroblastoma, osteosarcoma, normal cerebellum, or glioblastoma multiforme cell lines. By using a pair of primers recognizing a 280-bp region within the 630-bp 5' PTI-1 sequence, reverse transcription-PCR detects PTI-1 expression in patient-derived prostate carcinomas but not in normal prostate or benign hypertrophic prostate tissue. In contrast, reverse transcription-PCR detects prostate-specific antigen expression in all of the prostate tissues. These results indicate that PTI-1 may be a member of a class of oncogenes that could affect protein translation and contribute to carcinoma development in human prostate and other tissues. The approaches used, rapid expression cloning with the CREF-Trans 6 system and the DD strategy, should prove widely applicable for identifying and cloning additional human oncogenes.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adult
  • Amino Acid Sequence
  • Base Sequence
  • Carcinoma / genetics*
  • DNA, Complementary / genetics
  • Female
  • Gene Library
  • Humans
  • Male
  • Molecular Sequence Data
  • Oncogene Proteins / genetics*
  • Oncogenes / genetics*
  • Peptide Elongation Factor 1
  • Peptide Elongation Factors / genetics
  • Polymerase Chain Reaction
  • Prostatic Hyperplasia / genetics
  • Prostatic Neoplasms / genetics*
  • Sequence Homology, Amino Acid
  • Tissue Distribution
  • Tumor Cells, Cultured

Substances

  • DNA, Complementary
  • EEF1A1 protein, human
  • Oncogene Proteins
  • Peptide Elongation Factor 1
  • Peptide Elongation Factors

Associated data

  • GENBANK/L41490
  • GENBANK/L41498