title: Characterization of NOL7 gene point mutations, promoter methylation, and protein expression in cervical cancer.
abstract: NOL7 is a putative tumor suppressor gene localized to 6p23, a region with frequent loss of heterozygosity in a number of cancers, including cervical cancer (CC). We have previously demonstrated that reintroduction of NOL7 into CC cells altered the angiogenic phenotype and suppressed tumor growth in vivo by 95%. Therefore, to understand its mechanism of inactivation in CC, we investigated the genetic and epigenetic regulation of NOL7. NOL7 mRNA and protein levels were assessed in 13 CC cell lines and 23 consecutive CC specimens by real-time quantitative polymerase chain reaction, western blotting, and immunohistochemistry. Methylation of the NOL7 promoter was analyzed by bisulfite sequencing and mutations were identified through direct sequencing. A CpG island with multiple CpG dinucleotides spanned the 5' untranslated region and first exon of NOL7. However, bisulfite sequencing failed to identify persistent sites of methylation. Mutational sequencing revealed that 40% of the CC specimens and 31% of the CC cell lines harbored somatic mutations that may affect the in vivo function of NOL7. Endogenous NOL7 mRNA and protein expression in CC cell lines were significantly decreased in 46% of the CC cell lines. Finally, immunohistochemistry demonstrated strong NOL7 nucleolar staining in normal tissues that decreased with histologic progression toward CC. NOL7 is inactivated in CC in accordance with the Knudson 2-hit hypothesis through loss of heterozygosity and mutation. Together with evidence of its in vivo tumor suppression, these data support the hypothesis that NOL7 is the legitimate tumor suppressor gene located on 6p23.
authors: Department of Pathology, Medicine and Radiation and Cellular Oncology, University of Chicago, Chicago, IL 60637, USA.
authors: Colleen L
authors: CL
authors: Doçi
authors: Tanmayi P
authors: TP
authors: Mankame
authors: Alexander
authors: A
authors: Langerman
authors: Kelly R
authors: KR
authors: Ostler
authors: Rajani
authors: R
authors: Kanteti
authors: Timothy
authors: T
authors: Best
authors: Kenan
authors: K
authors: Onel
authors: Lucy A
authors: LA
authors: Godley
authors: Ravi
authors: R
authors: Salgia
authors: Mark W
authors: MW
authors: Lingen
mesh_terms: D045744:Cell Line, Tumor; D002896:Chromosomes, Human, Pair 6; D003001:Cloning, Molecular; D019175:DNA Methylation; D004252:DNA Mutational Analysis; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D020868:Gene Silencing; D006801:Humans; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism; D017354:Point Mutation; D011401:Promoter Regions, Genetic / Q000235:genetics* / Q000502:physiology; D060888:Real-Time Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D013447:Sulfites; D046888:Tissue Array Analysis; D025521:Tumor Suppressor Proteins / Q000235:genetics* / Q000378:metabolism; D002583:Uterine Cervical Neoplasms / Q000235:genetics* / Q000378:metabolism
chemical_list: C556669:NOL7 protein, human; D009687:Nuclear Proteins; D013447:Sulfites; D025521:Tumor Suppressor Proteins; C042345:hydrogen sulfite