title: Abnormalities of fragile histidine triad genomic and complementary DNAs in cervical cancer: association with human papillomavirus type.
abstract: BACKGROUND
  Chromosome 3p14.2 contains FRA3B, the most active chromosome breakage site in the human genome. The fragile histidine triad (FHIT) gene, a putative tumor suppressor gene, overlaps FRA3B. Human papillomavirus (HPV), a known cofactor in cervical carcinogenesis, can integrate into FRA3B. We examined abnormalities in FHIT and its RNA transcripts in cervical cancer cell lines and tumors. We also investigated the relationship between loss of heterozygosity (LOH) in FHIT/FRA3B and the presence of oncogenic HPV types.
  
  
  METHODS
  Eleven cell lines, 40 tumors (20 fresh and 20 archival), and 10 normal cervical epithelia were examined. Two intragenic polymorphic markers (D3S1300 and D3S4103) and the polymerase chain reaction (PCR) were used to examine FHIT LOH. Reverse transcription-PCR (RT-PCR) analysis and single-strand conformation polymorphism analysis of RT-PCR products were used to characterize FHIT transcripts. Oncogenic HPV types were identified by PCR, using general and type-specific primers.
  
  
  RESULTS
  All normal epithelia, 19 of 20 fresh tumors and nine of 11 cell lines expressed wild-type and, occasionally, exon 8-deleted FHIT transcripts. Additional aberrant FHIT transcripts were seen in nine of 20 fresh tumors and in seven of 11 cell lines. DNA sequencing of the aberrant transcripts revealed a variety of insertions and deletions but no point mutations. Three cell lines also had homozygous FHIT deletions. Oncogenic HPV types (i.e., 16, 18, 31, and 33) were detected in 18 of 20 archival tumors, and, in these tumors, LOH within FHIT was identified in nine of 16 informative cases. HPV 16 was found to be associated with LOH in the FHIT/FRA3B region (P = .041).
  
  
  CONCLUSIONS
  FHIT/FRA3B is frequently altered in cervical cancer, demonstrating LOH, occasional homozygous deletions, and frequent aberrant transcripts not found in normal epithelia. However, the presence of wild-type transcripts and the lack of protein-altering point mutations raise questions about FHIT's function as a classic tumor suppressor gene in cervical tissue.
authors.affiliation: Department of Obstetrics and Gynecology, Hamon Center for Therapeutic Oncology Research, The University of Texas Southwestern Medical Center, Dallas 75235-9032, USA. cmulle@mednet.swmed.edu
authors.forename: C Y
authors.initials: CY
authors.lastname: Muller
authors.forename: J D
authors.initials: JD
authors.lastname: O'Boyle
authors.forename: K M
authors.initials: KM
authors.lastname: Fong
authors.forename: I I
authors.initials: II
authors.lastname: Wistuba
authors.forename: E
authors.initials: E
authors.lastname: Biesterveld
authors.forename: M
authors.initials: M
authors.lastname: Ahmadian
authors.forename: D S
authors.initials: DS
authors.lastname: Miller
authors.forename: A F
authors.initials: AF
authors.lastname: Gazdar
authors.forename: J D
authors.initials: JD
authors.lastname: Minna
mesh_terms: D002872:Chromosome Deletion; D002873:Chromosome Fragility; D002893:Chromosomes, Human, Pair 3 / Q000235:genetics*; D018076:DNA, Complementary / Q000235:genetics*; D004273:DNA, Neoplasm / Q000235:genetics*; D005260:Female; D006579:Heterozygote; D006801:Humans; D027383:Papillomaviridae; D030361:Papillomavirus Infections / Q000150:complications*; D016133:Polymerase Chain Reaction / Q000379:methods; D018807:Polymorphism, Single-Stranded Conformational; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured; D014412:Tumor Virus Infections / Q000150:complications*; D002583:Uterine Cervical Neoplasms / Q000235:genetics* / Q000821:virology*
chemical_list: D018076:DNA, Complementary; D004273:DNA, Neoplasm