title: The candidate tumor suppressor gene, RASSF1A, from human chromosome 3p21.3 is involved in kidney tumorigenesis.
abstract: Clear cell-type renal cell carcinomas (clear RCC) are characterized almost universally by loss of heterozygosity on chromosome 3p, which usually involves any combination of three regions: 3p25-p26 (harboring the VHL gene), 3p12-p14.2 (containing the FHIT gene), and 3p21-p22, implying inactivation of the resident tumor-suppressor genes (TSGs). For the 3p21-p22 region, the affected TSGs remain, at present, unknown. Recently, the RAS association family 1 gene (isoform RASSF1A), located at 3p21.3, has been identified as a candidate lung and breast TSG. In this report, we demonstrate aberrant silencing by hypermethylation of RASSF1A in both VHL-caused clear RCC tumors and clear RCC without VHL inactivation. We found hypermethylation of RASSF1A's GC-rich putative promoter region in most of analyzed samples, including 39 of 43 primary tumors (91%). The promoter was methylated partially or completely in all 18 RCC cell lines analyzed. Methylation of the GC-rich putative RASSF1A promoter region and loss of transcription of the corresponding mRNA were related causally. RASSF1A expression was reactivated after treatment with 5-aza-2'-deoxycytidine. Forced expression of RASSF1A transcripts in KRC/Y, a renal carcinoma cell line containing a normal and expressed VHL gene, suppressed growth on plastic dishes and anchorage-independent colony formation in soft agar. Mutant RASSF1A had reduced growth suppression activity significantly. These data suggest that RASSF1A is the candidate renal TSG gene for the 3p21.3 region.
authors.affiliation: Laboratory of Immunobiology and Intramural Research Support Program, Science Applications International Corporation, National Cancer Institute, Frederick, MD 21702, USA.
authors.forename: K
authors.initials: K
authors.lastname: Dreijerink
authors.forename: E
authors.initials: E
authors.lastname: Braga
authors.forename: I
authors.initials: I
authors.lastname: Kuzmin
authors.forename: L
authors.initials: L
authors.lastname: Geil
authors.forename: F M
authors.initials: FM
authors.lastname: Duh
authors.forename: D
authors.initials: D
authors.lastname: Angeloni
authors.forename: B
authors.initials: B
authors.lastname: Zbar
authors.forename: M I
authors.initials: MI
authors.lastname: Lerman
authors.forename: E J
authors.initials: EJ
authors.lastname: Stanbridge
authors.forename: J D
authors.initials: JD
authors.lastname: Minna
authors.forename: A
authors.initials: A
authors.lastname: Protopopov
authors.forename: J
authors.initials: J
authors.lastname: Li
authors.forename: V
authors.initials: V
authors.lastname: Kashuba
authors.forename: G
authors.initials: G
authors.lastname: Klein
authors.forename: E R
authors.initials: ER
authors.lastname: Zabarovsky
mesh_terms: D001374:Azacitidine / Q000494:pharmacology; D002292:Carcinoma, Renal Cell / Q000235:genetics*; D002448:Cell Adhesion; D002455:Cell Division / Q000187:drug effects; D002874:Chromosome Mapping; D002893:Chromosomes, Human, Pair 3; D019175:DNA Methylation; D017931:DNA Primers; D004273:DNA, Neoplasm / Q000737:chemistry / Q000378:metabolism; D004318:Doxycycline / Q000633:toxicity; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D016147:Genes, Tumor Suppressor; D006801:Humans; D007680:Kidney Neoplasms / Q000235:genetics*; D009363:Neoplasm Proteins / Q000235:genetics* / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000378:metabolism; D015183:Restriction Mapping; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014407:Tumor Cells, Cultured; D025521:Tumor Suppressor Proteins
chemical_list: D017931:DNA Primers; D004273:DNA, Neoplasm; D009363:Neoplasm Proteins; C411762:RASSF1 protein, human; D011994:Recombinant Proteins; D025521:Tumor Suppressor Proteins; D001374:Azacitidine; D004318:Doxycycline