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Regulation of DNA repair gene expression in human cancer cell lines

Claire J McGurk1, Michele Cummings, Beate Köberle

  • 1Prostate Cancer Research Centre, Institute of Urology, UCL, 3rd Floor Research Laboratories, London, W1W 7EJ, United Kingdom.

Journal of Cellular Biochemistry
|November 30, 2005
PubMed

Insights

Testicular germ cell tumors are curable due to low nucleotide excision repair (NER) protein levels. This study found that low expression of ERCC1, XPF, and XPA DNA repair proteins in these tumors is controlled at the translational level.

Area of Science:

  • Molecular Biology
  • Cancer Research
  • Genetics

Background:

  • Most advanced cancers are incurable, but testicular germ cell tumors (TGCTs) are highly curable with cisplatin chemotherapy.
  • The nucleotide excision repair (NER) pathway removes DNA adducts formed by cisplatin, and its low activity in TGCTs correlates with curability.
  • Key NER proteins ERCC1, XPF, and XPA are expressed at low levels in TGCT cell lines.

Purpose of the Study:

  • To investigate the mechanism underlying the reduced expression of ERCC1, XPF, and XPA proteins in TGCTs.
  • To determine if transcriptional or post-transcriptional mechanisms explain the low DNA repair protein levels in testis tumor cells.

Main Methods:

  • Real-time PCR was used to quantify mRNA levels of ERCC1, XPF, and XPA in 14 human cancer cell lines.
  • Analysis included identification and quantification of ERCC1 splice variants and transcription start points (TSPs).
  • Pulse-chase experiments assessed the protein half-life of ERCC1 in TGCT and prostate cancer cell lines.

Main Results:

  • Significantly lower protein levels of ERCC1, XPF, and XPA were observed in TGCT cell lines compared to others.
  • Transcriptional efficiency and mRNA stability did not fully explain the reduced protein levels.
  • Alternative splicing of ERCC1 did not account for the testis-specific low protein expression.
  • ERCC1 protein half-life was similar in TGCT and prostate cancer cell lines, ruling out post-translational degradation differences.

Conclusions:

  • The low constitutive levels of ERCC1, XPF, and XPA proteins in TGCTs are not explained by transcriptional regulation or mRNA stability.
  • Post-transcriptional regulation via alternative splicing or protein degradation does not account for the observed low expression.
  • Evidence suggests that the control of these crucial DNA repair proteins in TGCTs occurs at the translational level.

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