Identification of the human prostatic carcinoma oncogene PTI-1 by rapid expression cloning and differential RNA display.
- 18 July 1995
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 92 (15) , 6778-6782
- https://doi.org/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.Keywords
This publication has 32 references indexed in Scilit:
- The melanoma differentiation-associated gene mda-6, which encodes the cyclin-dependent kinase inhibitor p21, is differentially expressed during growth, differentiation and progression in human melanoma cells.1995
- Chromosomal translocations in human cancerNature, 1994
- Microsatellite instability: marker of a mutator phenotype in cancer.1994
- Correlation of pathologic findings with progression after radical retropubic prostatectomyCancer, 1993
- An assessment of radical prostatectomy. Time trends, geographic variation, and outcomes. The Prostate Patient Outcomes Research TeamPublished by American Medical Association (AMA) ,1993
- Computerized Tomography and Transrectal Ultrasound in the Assessment of Local Extension of Prostatic Cancer Before Radical Retropubic ProstatectomyJournal of Urology, 1987
- Mutant EF-Tu increases missense error in vitroMolecular Genetics and Genomics, 1986
- Changes in activity and amount of active elongation factor 1α in aging and immortal human fibroblast culturesExperimental Gerontology, 1986
- New human transforming genes detected by a tumorigenicity assay.Molecular and Cellular Biology, 1984
- LNCAP MODEL OF HUMAN PROSTATIC-CARCINOMA1983