Restoration of endogenous wild-type p53 activity in a glioblastoma cell line with intrinsic temperature-sensitive p53 induces growth arrest but not apoptosis

Jun Ikeda, Mitsuhiro Tada, Nobuaki Ishii, Hideyuki Saya, Kazuhiko Tsuchiya, Kumio Okaichi, Kazuhiko Mishima, Yutaka Sawamura, Giulia Fulci, Ta Jen Liu, Erwin G. Van Meir

研究成果: Article

11 引用 (Scopus)

抄録

p53 protein is a transcription factor involved in multiple tumor-suppressor activities including cell cycle control and apoptosis. TP53 gene is frequently mutated in glioblastoma, suggesting the importance of inactivation of this gene product in gliomagenesis. Restoration of p53 function in glioblastoma cell lines deficient for p53 has shown that p53 induces growth arrest or apoptosis depending on the cell line and vector used to transduce wild-type TP53 alleles. Considering that astrocytes grow and express p53, it is not clear whether these results reflect physiologic responses or the result of p53 overexpression in combination with cellular responses to viral vector infection. Here, we reassessed this issue using a glioblastoma cell line (LN382) that expresses an endogenous temperature-sensitive mutant p53. This cell line expresses TP53 alleles (100% as determined by a p53 transcriptional assay in yeast) mutated at codon 197 GTG (Val) > CTG (Leu). We found that the p53 protein in these cells acted as an inactive mutant at 37°C and as a functional wild-type p53 below 34°C as demonstrated by several lines of evidence, including (i) restoration of transactivating ability in yeast, (ii) induction of p53-modulated genes such as CDKN1p21 and transforming growth factor-α, (iii) disappearance of accumulated p53 protein in the nucleus and (iv) decrease in steady state p53 protein levels. This temperature switch allowed p53 levels, which were close to physiological levels to dramatically reduce LN382 cell proliferation by inducing a G1/S cell cycle block, but not to induce apoptosis. The lack of apoptosis was considered to be a result of the low level p53 expression, because increasing wild-type p53 levels by adenoviral-mediated gene transfer caused apoptosis in these cells. The LN382 cell line will be extremely useful for investigations into the roles of p53 in cellular responses to a variety of stimuli or damages.

元の言語English
ページ(範囲)35-43
ページ数9
ジャーナルInternational Journal of Cancer
94
発行部数1
DOI
出版物ステータスPublished - 2001 10 1
外部発表Yes

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Glioblastoma
Apoptosis
Cell Line
Temperature
Growth
p53 Genes
Proteins
Yeasts
Alleles
Transforming Growth Factors
Gene Silencing
Virus Diseases
Cell Cycle Checkpoints
Codon
Astrocytes
Cell Cycle
Transcription Factors
Cell Proliferation
Genes
Neoplasms

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

これを引用

Restoration of endogenous wild-type p53 activity in a glioblastoma cell line with intrinsic temperature-sensitive p53 induces growth arrest but not apoptosis. / Ikeda, Jun; Tada, Mitsuhiro; Ishii, Nobuaki; Saya, Hideyuki; Tsuchiya, Kazuhiko; Okaichi, Kumio; Mishima, Kazuhiko; Sawamura, Yutaka; Fulci, Giulia; Liu, Ta Jen; Van Meir, Erwin G.

:: International Journal of Cancer, 巻 94, 番号 1, 01.10.2001, p. 35-43.

研究成果: Article

Ikeda, J, Tada, M, Ishii, N, Saya, H, Tsuchiya, K, Okaichi, K, Mishima, K, Sawamura, Y, Fulci, G, Liu, TJ & Van Meir, EG 2001, 'Restoration of endogenous wild-type p53 activity in a glioblastoma cell line with intrinsic temperature-sensitive p53 induces growth arrest but not apoptosis', International Journal of Cancer, 巻. 94, 番号 1, pp. 35-43. https://doi.org/10.1002/ijc.1431
Ikeda, Jun ; Tada, Mitsuhiro ; Ishii, Nobuaki ; Saya, Hideyuki ; Tsuchiya, Kazuhiko ; Okaichi, Kumio ; Mishima, Kazuhiko ; Sawamura, Yutaka ; Fulci, Giulia ; Liu, Ta Jen ; Van Meir, Erwin G. / Restoration of endogenous wild-type p53 activity in a glioblastoma cell line with intrinsic temperature-sensitive p53 induces growth arrest but not apoptosis. :: International Journal of Cancer. 2001 ; 巻 94, 番号 1. pp. 35-43.
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abstract = "p53 protein is a transcription factor involved in multiple tumor-suppressor activities including cell cycle control and apoptosis. TP53 gene is frequently mutated in glioblastoma, suggesting the importance of inactivation of this gene product in gliomagenesis. Restoration of p53 function in glioblastoma cell lines deficient for p53 has shown that p53 induces growth arrest or apoptosis depending on the cell line and vector used to transduce wild-type TP53 alleles. Considering that astrocytes grow and express p53, it is not clear whether these results reflect physiologic responses or the result of p53 overexpression in combination with cellular responses to viral vector infection. Here, we reassessed this issue using a glioblastoma cell line (LN382) that expresses an endogenous temperature-sensitive mutant p53. This cell line expresses TP53 alleles (100{\%} as determined by a p53 transcriptional assay in yeast) mutated at codon 197 GTG (Val) > CTG (Leu). We found that the p53 protein in these cells acted as an inactive mutant at 37°C and as a functional wild-type p53 below 34°C as demonstrated by several lines of evidence, including (i) restoration of transactivating ability in yeast, (ii) induction of p53-modulated genes such as CDKN1p21 and transforming growth factor-α, (iii) disappearance of accumulated p53 protein in the nucleus and (iv) decrease in steady state p53 protein levels. This temperature switch allowed p53 levels, which were close to physiological levels to dramatically reduce LN382 cell proliferation by inducing a G1/S cell cycle block, but not to induce apoptosis. The lack of apoptosis was considered to be a result of the low level p53 expression, because increasing wild-type p53 levels by adenoviral-mediated gene transfer caused apoptosis in these cells. The LN382 cell line will be extremely useful for investigations into the roles of p53 in cellular responses to a variety of stimuli or damages.",
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AU - Tada, Mitsuhiro

AU - Ishii, Nobuaki

AU - Saya, Hideyuki

AU - Tsuchiya, Kazuhiko

AU - Okaichi, Kumio

AU - Mishima, Kazuhiko

AU - Sawamura, Yutaka

AU - Fulci, Giulia

AU - Liu, Ta Jen

AU - Van Meir, Erwin G.

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AB - p53 protein is a transcription factor involved in multiple tumor-suppressor activities including cell cycle control and apoptosis. TP53 gene is frequently mutated in glioblastoma, suggesting the importance of inactivation of this gene product in gliomagenesis. Restoration of p53 function in glioblastoma cell lines deficient for p53 has shown that p53 induces growth arrest or apoptosis depending on the cell line and vector used to transduce wild-type TP53 alleles. Considering that astrocytes grow and express p53, it is not clear whether these results reflect physiologic responses or the result of p53 overexpression in combination with cellular responses to viral vector infection. Here, we reassessed this issue using a glioblastoma cell line (LN382) that expresses an endogenous temperature-sensitive mutant p53. This cell line expresses TP53 alleles (100% as determined by a p53 transcriptional assay in yeast) mutated at codon 197 GTG (Val) > CTG (Leu). We found that the p53 protein in these cells acted as an inactive mutant at 37°C and as a functional wild-type p53 below 34°C as demonstrated by several lines of evidence, including (i) restoration of transactivating ability in yeast, (ii) induction of p53-modulated genes such as CDKN1p21 and transforming growth factor-α, (iii) disappearance of accumulated p53 protein in the nucleus and (iv) decrease in steady state p53 protein levels. This temperature switch allowed p53 levels, which were close to physiological levels to dramatically reduce LN382 cell proliferation by inducing a G1/S cell cycle block, but not to induce apoptosis. The lack of apoptosis was considered to be a result of the low level p53 expression, because increasing wild-type p53 levels by adenoviral-mediated gene transfer caused apoptosis in these cells. The LN382 cell line will be extremely useful for investigations into the roles of p53 in cellular responses to a variety of stimuli or damages.

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