PMID- 12779080
OWN - NLM
STAT- MEDLINE
DCOM- 20040123
LR  - 20190906
IS  - 0167-6806 (Print)
IS  - 0167-6806 (Linking)
VI  - 79
IP  - 1
DP  - 2003 May
TI  - Comparison of p53 mutational status with mRNA and protein expression in a panel 
      of 24 human breast carcinoma cell lines.
PG  - 37-46
AB  - We analyzed the p53 mutational status, mRNA and protein expression in 24 human 
      breast carcinoma cell lines. Following measurement of their DNA content with flow 
      cytometry, we ascertained the copy numbers of the centromere of chromosome 17 
      (cen17) and p53 with fluorescence in situ hybridization (FISH). A functional 
      yeast assay (FASAY) was used to screen for inactivating mutations. Positive 
      results were subsequently verified by DNA sequencing. Finally, we assessed the 
      mRNA expression with a competitive reverse transcription-polymerase chain 
      reaction (RT-PCR) assay and the protein expression with immunocytochemical 
      staining, western blot, and quantitative flow cytometry. The DNA content of the 
      cell lines ranged from 0.85 to 2.58. Nine cell lines had concordant copy numbers 
      (between two and four) of p53 and cen17, whereas 12 had more, and three less 
      cen17 than p53 copies. The FASAY was successful in all but one cell line and 
      revealed the presence of mutated alleles in 16 of them, 13 cell lines expressed 
      only the mutated, and three both the mutated and the wild-type alleles. The 
      mutations were comprised of 11 missense, two nonsense, and three frameshift 
      mutations. Immunocytochemical staining, western blot and quantitative flow 
      cytometry yielded comparable p53 protein expression results. However, both the 
      mRNA and the protein expression levels varied considerably in the different cell 
      lines and no consistent pattern with regard to the respective p53 mutational 
      status became evident. The results obtained in these breast carcinoma cell lines 
      indicate that no clear-cut linear relationship exists between the p53 mutational 
      status and the extent of its respective mRNA and protein expression. Therefore, 
      direct DNA analyses and functional assays remain the only methods for the 
      reliable detection of p53 mutations.
FAU - Concin, Nicole
AU  - Concin N
AD  - Molecular Oncology Group, Department of Gynecology and Obstetrics, University of 
      Vienna Medical School, Vienna, Austria.
FAU - Zeillinger, Christa
AU  - Zeillinger C
FAU - Tong, Dan
AU  - Tong D
FAU - Stimpfl, Margit
AU  - Stimpfl M
FAU - Konig, Margit
AU  - Konig M
FAU - Printz, Dieter
AU  - Printz D
FAU - Stonek, Felix
AU  - Stonek F
FAU - Schneeberger, Christian
AU  - Schneeberger C
FAU - Hefler, Lukas
AU  - Hefler L
FAU - Kainz, Christian
AU  - Kainz C
FAU - Leodolter, Sepp
AU  - Leodolter S
FAU - Haas, Oskar A
AU  - Haas OA
FAU - Zeillinger, Robert
AU  - Zeillinger R
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Netherlands
TA  - Breast Cancer Res Treat
JT  - Breast cancer research and treatment
JID - 8111104
RN  - 0 (DNA, Neoplasm)
RN  - 0 (RNA, Messenger)
RN  - 0 (Tumor Suppressor Protein p53)
SB  - IM
MH  - Breast Neoplasms/*genetics/metabolism
MH  - Carcinoma/*genetics/metabolism
MH  - Centromere/genetics
MH  - Chromosomes, Human, Pair 17/genetics
MH  - DNA Mutational Analysis
MH  - DNA, Neoplasm/*genetics
MH  - Gene Expression
MH  - Genes, p53/*genetics
MH  - Humans
MH  - Immunohistochemistry
MH  - In Situ Hybridization, Fluorescence
MH  - *Mutation
MH  - RNA, Messenger/analysis
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Tumor Cells, Cultured
MH  - Tumor Suppressor Protein p53/genetics/metabolism
EDAT- 2003/06/05 05:00
MHDA- 2004/01/24 05:00
CRDT- 2003/06/05 05:00
PHST- 2003/06/05 05:00 [pubmed]
PHST- 2004/01/24 05:00 [medline]
PHST- 2003/06/05 05:00 [entrez]
AID - 10.1023/a:1023351717408 [doi]
PST - ppublish
SO  - Breast Cancer Res Treat. 2003 May;79(1):37-46. doi: 10.1023/a:1023351717408.