PMID- 15884040
OWN - NLM
STAT- MEDLINE
DCOM- 20061215
LR  - 20091119
IS  - 1098-1004 (Electronic)
IS  - 1059-7794 (Linking)
VI  - 25
IP  - 6
DP  - 2005 Jun
TI  - Sensitive multistep clinical molecular screening of 180 unrelated individuals 
      with retinoblastoma detects 36 novel mutations in the RB1 gene.
PG  - 566-74
AB  - Retinoblastoma (RB) is a neoplasm of retinal origin caused by mutations in RB1, 
      the retinoblastoma tumor suppressor gene. To facilitate genetics counseling and 
      patient management, we adopted a multistep molecular screening assay for 
      detecting RB1 mutations. This assay included DNA sequencing to identify mutations 
      within coding exons and immediate flanking intronic regions, Southern blot 
      analysis to characterize genomic rearrangements, and transcript analysis to 
      characterize potential splicing mutations buried within introns. In a pilot 
      investigation of 180 patients from North America, we identified germline RB1 
      mutations in 77 out of 85 bilateral RB patients (91%), 7 out of 10 familial 
      unilateral (70%), and 6 out of 85 unilateral patients with no family history of 
      RB (7%). Mutations included 36 novel alterations spanning the entire RB1 gene. 
      Seven of these novel changes were missense or silent mutations. Sequence analysis 
      predicted that, in five out of seven cases, the changes can cause aberrant 
      splicing. This was confirmed by transcript analysis in four out of five cases. In 
      addition, four intronic point mutations within nonconsensus sites activated 
      cryptic splice sites. Without the transcript analysis, the significance of these 
      11 mutations would have remained undefined. In a separate investigation of a 
      subset of unilateral RB tumors, we identified somatic biallelic RB1 gene 
      inactivation in 34 out of 56 cases (61%) cases. In 14 tumors, only one of the two 
      RB1 mutations could be detected, and in eight tumors, no mutations were detected. 
      The absence of detectable RB1 mutations in eight bilateral cases and eight 
      unilateral tumors suggests that alternative genetic mechanisms may underlie the 
      development of RB in certain individuals.
FAU - Nichols, Kim E
AU  - Nichols KE
AD  - Division of Pediatric Oncology, Children's Hospital of Philadelphia, 
      Philadelphia, Pennsylvania, USA.
FAU - Houseknecht, Monisa D
AU  - Houseknecht MD
FAU - Godmilow, Lynn
AU  - Godmilow L
FAU - Bunin, Greta
AU  - Bunin G
FAU - Shields, Carol
AU  - Shields C
FAU - Meadows, Anna
AU  - Meadows A
FAU - Ganguly, Arupa
AU  - Ganguly A
LA  - eng
SI  - GENBANK/L11910
SI  - OMIM/180200
SI  - RefSeq/NM_000321
GR  - CA81012/CA/NCI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Hum Mutat
JT  - Human mutation
JID - 9215429
RN  - 0 (RNA, Messenger)
RN  - 0 (Retinoblastoma Protein)
SB  - IM
MH  - DNA Mutational Analysis
MH  - Exons/genetics
MH  - Genetic Testing/*methods
MH  - Genome, Human/genetics
MH  - Germ-Line Mutation/*genetics
MH  - Humans
MH  - Molecular Sequence Data
MH  - Polymorphism, Genetic
MH  - RNA, Messenger/genetics/metabolism
MH  - Retinoblastoma/*genetics
MH  - Retinoblastoma Protein/*genetics
MH  - Sensitivity and Specificity
EDAT- 2005/05/11 09:00
MHDA- 2006/12/16 09:00
CRDT- 2005/05/11 09:00
PHST- 2005/05/11 09:00 [pubmed]
PHST- 2006/12/16 09:00 [medline]
PHST- 2005/05/11 09:00 [entrez]
AID - 10.1002/humu.20184 [doi]
PST - ppublish
SO  - Hum Mutat. 2005 Jun;25(6):566-74. doi: 10.1002/humu.20184.