P21Cip1 is a critical mediator of the cytotoxic action of thymidylate synthase inhibitors in colorectal carcinoma cells

James I. Geller, Kinga Szekely-Szucs, I. Peták, Belinda Doyle, Janet A. Houghton

Research output: Contribution to journalArticle

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Abstract

We have demonstrated previously that interferon (IFN)-,γ sensitizes human colon carcinoma cell lines to the cytotoxic effects of 5-fluorouracil combined with leucovorin and to the thymidylate synthase inhibitor, ZD9331, dependent on thymineless stress-induced DNA damage, independent of p53. Here we demonstrate that the cyclin-dependent kinase (CDK) inhibitor p21Cip1 regulates thymineless stress-induced cytotoxicity in these cells. HCT116 wild-type (wt) and p53-/- cells underwent apoptosis and loss in clonogenic survival when exposed to ZD9331, whereas p21Cip1 P-/- cells were resistant. In contrast, IFN-γ induced marked cytotoxicity in p21 Cip1-/- cells only. ZD9331 induced p21Cip1 up-regulation in all of the cell lines examined, as did thymidine deprivation in thymidylate synthase-deficient (thymidylate synthase-) cells. Furthermore, selective induction of p21Cip1 in RKO was sufficient to induce apoptosis. P21Cip1, cdk1, cdk2, and cyclin E mRNA expression increased coincident with S-phase accumulation in HT29 cells treated with ZD9331 or 5-fluorouracil/leucovorin, as demonstrated by cDNA microarray analyses. Cell cycle analyses revealed that HCT116 wt and p21Cip1 -/- cells accumulated in S phase within 24 h of ZD9331 exposure; however, wt cells exited S-phase more rapidly, where apoptosis occurred before mitosis, either in late S or G2. Finally, the CDK inhibitor roscovitine potentiated the cytotoxic activity of ZD9331 in both wt and p21Cip1-/- cells, strongly suggesting a role for p21Cip1-dependent CDK inhibition in cytotoxicity induced by thymidylate synthase inhibition. In summary, p21 Cip1 positively regulates the cytotoxic action of thymidylate synthase inhibitors, negatively regulates the cytotoxic action of IFN-γ, and enhances S-phase exit after thymineless stress, possibly via interaction with CDK-cyclin complexes.

Original languageEnglish
Pages (from-to)6296-6303
Number of pages8
JournalCancer Research
Volume64
Issue number17
DOIs
Publication statusPublished - Sep 1 2004

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Thymidylate Synthase
Colorectal Neoplasms
Cyclin-Dependent Kinases
S Phase
Interferons
Leucovorin
Apoptosis
Fluorouracil
Cell Line
Cyclin E
HT29 Cells
Cyclins
Microarray Analysis
Oligonucleotide Array Sequence Analysis
Mitosis
Thymidine
DNA Damage
ZD 9331
Cell Cycle
Colon

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

P21Cip1 is a critical mediator of the cytotoxic action of thymidylate synthase inhibitors in colorectal carcinoma cells. / Geller, James I.; Szekely-Szucs, Kinga; Peták, I.; Doyle, Belinda; Houghton, Janet A.

In: Cancer Research, Vol. 64, No. 17, 01.09.2004, p. 6296-6303.

Research output: Contribution to journalArticle

Geller, James I. ; Szekely-Szucs, Kinga ; Peták, I. ; Doyle, Belinda ; Houghton, Janet A. / P21Cip1 is a critical mediator of the cytotoxic action of thymidylate synthase inhibitors in colorectal carcinoma cells. In: Cancer Research. 2004 ; Vol. 64, No. 17. pp. 6296-6303.
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AU - Szekely-Szucs, Kinga

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AU - Doyle, Belinda

AU - Houghton, Janet A.

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