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Expression of C-terminal deleted p53 isoforms in neuroblastoma.

Goldschneider D, Horvilleur E, Plassa LF, Guillaud-Bataille M, Million K, Wittmer-Dupret E, Danglot G, de Thé H, Bénard J, May E, Douc-Rasy S - Nucleic Acids Res. (2006)

Bottom Line: Xirodimas, M.Saville and D.Lane (2005) Genes Dev., 19, 2122-2137].

View Article: PubMed Central - PubMed

Affiliation: Centre National de Recherche Scientifique, UMR 8126, Institut Gustave Roussy, 94805 Villejuif, France.

ABSTRACT
The tumor suppressor gene, p53, is rarely mutated in neuroblastomas (NB) at the time of diagnosis, but its dysfunction could result from a nonfunctional conformation or cytoplasmic sequestration of the wild-type p53 protein. However, p53 mutation, when it occurs, is found in NB tumors with drug resistance acquired over the course of chemotherapy. As yet, no study has been devoted to the function of the specific p53 mutants identified in NB cells. This study includes characterization and functional analysis of p53 expressed in eight cell lines: three wild-type cell lines and five cell lines harboring mutations. We identified two transcription-inactive p53 variants truncated in the C-terminus, one of which corresponded to the p53beta isoform recently identified in normal tissue by Bourdon et al. [J. C. Bourdon, K. Fernandes, F. Murray-Zmijewski, G. Liu, A. Diot, D. P. Xirodimas, M. K. Saville and D. P. Lane (2005) Genes Dev., 19, 2122-2137]. Our results show, for the first time, that the p53beta isoform is the only p53 species to be endogenously expressed in the human NB cell line SK-N-AS, suggesting that the C-terminus truncated p53 isoforms may play an important role in NB tumor development.

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Related in: MedlinePlus

Identification of a deletion spanning the intron9/exon 10 junction of SK-N-AS p53 gene. (A) Schematic representation of p53 gene from intron 7 to intron 10 with the position of amplified fragments; (B) PCR fragments amplified from SH-SY5Y (SH) and SK-N-AS (AS) DNA with the primer pairs a, b, c and d. The primer sequences are given in Table 1; contl: PCR performed in parallel without DNA.
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fig3: Identification of a deletion spanning the intron9/exon 10 junction of SK-N-AS p53 gene. (A) Schematic representation of p53 gene from intron 7 to intron 10 with the position of amplified fragments; (B) PCR fragments amplified from SH-SY5Y (SH) and SK-N-AS (AS) DNA with the primer pairs a, b, c and d. The primer sequences are given in Table 1; contl: PCR performed in parallel without DNA.

Mentions: A series of genomic amplifications were performed to identify a possible deletion within the intron 9 that could account for the absence of normal size p53 in SK-N-AS cells. The primer sequences are given in Table 1. Amplifications were performed in parallel with total DNA extracted from SH-SY5Y and SK-N-AS cells. Results are presented in Figure 3. Normal size fragments that encompass the acceptor site of intron 9 were amplified with SH-SY5Y as well as with SK-N-AS DNA. On the contrary amplification fragments that encompass the intron 9 donor site were obtained only with SH-SY5Y but not with SK-N-AS DNA. These results identify a deletion of the intron 9/exon 10 junction within the SK-N-AS p53 gene.


Expression of C-terminal deleted p53 isoforms in neuroblastoma.

Goldschneider D, Horvilleur E, Plassa LF, Guillaud-Bataille M, Million K, Wittmer-Dupret E, Danglot G, de Thé H, Bénard J, May E, Douc-Rasy S - Nucleic Acids Res. (2006)

Identification of a deletion spanning the intron9/exon 10 junction of SK-N-AS p53 gene. (A) Schematic representation of p53 gene from intron 7 to intron 10 with the position of amplified fragments; (B) PCR fragments amplified from SH-SY5Y (SH) and SK-N-AS (AS) DNA with the primer pairs a, b, c and d. The primer sequences are given in Table 1; contl: PCR performed in parallel without DNA.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC1636465&req=5

fig3: Identification of a deletion spanning the intron9/exon 10 junction of SK-N-AS p53 gene. (A) Schematic representation of p53 gene from intron 7 to intron 10 with the position of amplified fragments; (B) PCR fragments amplified from SH-SY5Y (SH) and SK-N-AS (AS) DNA with the primer pairs a, b, c and d. The primer sequences are given in Table 1; contl: PCR performed in parallel without DNA.
Mentions: A series of genomic amplifications were performed to identify a possible deletion within the intron 9 that could account for the absence of normal size p53 in SK-N-AS cells. The primer sequences are given in Table 1. Amplifications were performed in parallel with total DNA extracted from SH-SY5Y and SK-N-AS cells. Results are presented in Figure 3. Normal size fragments that encompass the acceptor site of intron 9 were amplified with SH-SY5Y as well as with SK-N-AS DNA. On the contrary amplification fragments that encompass the intron 9 donor site were obtained only with SH-SY5Y but not with SK-N-AS DNA. These results identify a deletion of the intron 9/exon 10 junction within the SK-N-AS p53 gene.

Bottom Line: Xirodimas, M.Saville and D.Lane (2005) Genes Dev., 19, 2122-2137].

View Article: PubMed Central - PubMed

Affiliation: Centre National de Recherche Scientifique, UMR 8126, Institut Gustave Roussy, 94805 Villejuif, France.

ABSTRACT
The tumor suppressor gene, p53, is rarely mutated in neuroblastomas (NB) at the time of diagnosis, but its dysfunction could result from a nonfunctional conformation or cytoplasmic sequestration of the wild-type p53 protein. However, p53 mutation, when it occurs, is found in NB tumors with drug resistance acquired over the course of chemotherapy. As yet, no study has been devoted to the function of the specific p53 mutants identified in NB cells. This study includes characterization and functional analysis of p53 expressed in eight cell lines: three wild-type cell lines and five cell lines harboring mutations. We identified two transcription-inactive p53 variants truncated in the C-terminus, one of which corresponded to the p53beta isoform recently identified in normal tissue by Bourdon et al. [J. C. Bourdon, K. Fernandes, F. Murray-Zmijewski, G. Liu, A. Diot, D. P. Xirodimas, M. K. Saville and D. P. Lane (2005) Genes Dev., 19, 2122-2137]. Our results show, for the first time, that the p53beta isoform is the only p53 species to be endogenously expressed in the human NB cell line SK-N-AS, suggesting that the C-terminus truncated p53 isoforms may play an important role in NB tumor development.

Show MeSH
Related in: MedlinePlus