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Molecular Biology and Genetics |
Intramural Research Support Program, SAIC-Frederick, Inc. [I. K., L. G., F-M. D.] and Laboratory of Immunobiology [K. D., M. I. L.], National Cancer Institute Frederick, Frederick, Maryland 21702; Laboratory of Pathology [J. W. G., M. R. E-B.] and Urologic Oncology Branch [Y. Y., C. D. V., W. M. L.], National Cancer Institute Bethesda, Bethesda, Maryland 20892; MTC and CGR, The Karolinska Institute, 171 77 Stockholm, Sweden [A. P., E. Z.]; Engelhardt Institute of Molecular Biology, Moscow 117984, Russia [E. Z.]; Hamon Center for Therapeutic Oncology Research, University of Texas Southwestern Medical Center, Dallas, TX 75390 [J. D. M.]; and Center for Prostate Disease Research, Uniformed Services University of Health Sciences, Bethesda, Maryland, 20814 [J. S. R.]
We analyzed expression status of the recently identified tumor suppressor geneRASSF1A in primary prostate carcinomas and in prostate cell lines. We found complete methylation of the RASSF1A promoter in 63% of primary microdissected prostate carcinomas (7 of 11 samples). The remaining 4 samples (37%) were partially methylated, possibly because of contamination with normal cells. No promoter methylation was observed in matching normal prostate tissues. High levels of RASSF1A transcript and no methylation of RASSF1A promoter were found in explanted primary normal prostate epithelial and stromal cells. Complete silencing and methylation of RASSF1A promoter was observed in five widely used prostate carcinoma cell lines, which acquired the ability to grow in culture spontaneously, including LNCaP, PC-3, ND-1, DU-145, 22Rv1, and one primary prostate carcinoma immortalized by overexpression of the human telomerase catalytic subunit (RC-58T/hTERT). However, no silencing of RASSF1A was found in four other prostate carcinoma cell lines, which were adapted for cell culture after transformation with human papillomaviral DNA. Suppression of cell growth in vitro was demonstrated after the reintroduction of RASSF1A-expressing construct into LNCaP prostate carcinoma cells. Our data implicate the RASSF1A gene in human prostate tumorigenesis.
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