Modulation of CDK2-AP1 (p12DOC-1) expression in human colorectal cancer

Ziquan Yuan, Anu G. Gaba, Tara Sotsky Kent, Anna Bennett, Agnes Miller, Thomas K. Weber

Research output: Contribution to journalArticle

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Abstract

We have previously demonstrated an association between microsatellite instability and decreased CDK2-AP1 (p12DOC-1) expression in human colorectal cancer (CRC) cell lines. In those same studies, induction of CDK2-AP1 expression promoted both cell cycle arrest and apoptosis. The goals of our present study were to better understand the mechanisms leading to reduced CDK2-AP1 expression in microsatellite unstable (MSI) CRC and to study further the effect of CDK2-AP1 modulation on cell proliferation and apoptosis utilizing RNA interference (RNAi) techniques. We used direct sequencing to screen for mutations of the poly (T)8 microsatellite-like region in the 3′ end of the CDK2-AP1 gene in 24 CRC cell lines. We then utilized an in vitro human mismatch repair (MMR) recombinant system to assess for correction of the mutation and changes in CDK2-AP1 expression secondary to hMLH1 transfection. We also investigated the effect of CDK2-AP1 modulation in four settings: (1) native CDK2-AP1 absence, (2) endogenous CDK2-AP1 expression, (3) RNAi-induced CDK2-AP1 inhibition and (4) induced CDK2-AP1 over expression. The mutation - del T poly (T)8 - at the 3′ end of the CDK2-AP1 gene was found in 3/12 (25%) of MSI CRC cell lines, but in none of the microsatellite stable samples (0/12). Interestingly, when wild-type MMR protein - MLH1 - was induced in an in vitro human recombinant system, the del T poly (T)8 mutation was reversed and CDK2-AP1 expression increased. RNAi-mediated CDK2-AP1 inhibition was associated with decreased apoptosis and increased cell proliferation in CDK2-AP1-non deficient CRC cell lines. We conclude that mutations in the microsatellite-like sequence of the CDK2-AP1 gene in MSI CRC are associated with decreased CDK2-AP1 expression. In addition, modulation of CDK2-AP1 expression in human CRC alters cell proliferation and apoptosis.

Original languageEnglish (US)
Pages (from-to)3657-3668
Number of pages12
JournalOncogene
Volume24
Issue number22
DOIs
StatePublished - May 19 2005

Fingerprint

Colorectal Neoplasms
Microsatellite Repeats
Poly T
RNA Interference
Mutation
Apoptosis
Cell Line
DNA Mismatch Repair
Cell Proliferation
Genes
Microsatellite Instability
Cell Cycle Checkpoints
Transfection

Keywords

  • CDK2-AP1
  • Colorectal cancer
  • Microsatellite instability
  • siRNA

ASJC Scopus subject areas

  • Molecular Biology
  • Cancer Research
  • Genetics

Cite this

Yuan, Z., Gaba, A. G., Kent, T. S., Bennett, A., Miller, A., & Weber, T. K. (2005). Modulation of CDK2-AP1 (p12DOC-1) expression in human colorectal cancer. Oncogene, 24(22), 3657-3668. https://doi.org/10.1038/sj.onc.1208378

Modulation of CDK2-AP1 (p12DOC-1) expression in human colorectal cancer. / Yuan, Ziquan; Gaba, Anu G.; Kent, Tara Sotsky; Bennett, Anna; Miller, Agnes; Weber, Thomas K.

In: Oncogene, Vol. 24, No. 22, 19.05.2005, p. 3657-3668.

Research output: Contribution to journalArticle

Yuan, Z, Gaba, AG, Kent, TS, Bennett, A, Miller, A & Weber, TK 2005, 'Modulation of CDK2-AP1 (p12DOC-1) expression in human colorectal cancer', Oncogene, vol. 24, no. 22, pp. 3657-3668. https://doi.org/10.1038/sj.onc.1208378
Yuan Z, Gaba AG, Kent TS, Bennett A, Miller A, Weber TK. Modulation of CDK2-AP1 (p12DOC-1) expression in human colorectal cancer. Oncogene. 2005 May 19;24(22):3657-3668. https://doi.org/10.1038/sj.onc.1208378
Yuan, Ziquan ; Gaba, Anu G. ; Kent, Tara Sotsky ; Bennett, Anna ; Miller, Agnes ; Weber, Thomas K. / Modulation of CDK2-AP1 (p12DOC-1) expression in human colorectal cancer. In: Oncogene. 2005 ; Vol. 24, No. 22. pp. 3657-3668.
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