Increased expression of T-fimbrin gene after DNA damage in CHO cells and inactivation of T-fimbrin by CpG methylation in human colorectal cancer cells

Int J Cancer. 2002 Jan 10;97(2):211-6. doi: 10.1002/ijc.1587.

Abstract

When DNA damage is induced by unprogrammed extrinsic events, activating-cell-cycle checkpoints delay cell-cycle progression in the G1 or G2 phases and allow repair of a damaged template. In this study, we evaluated changes in gene expression upon radiation-induced G2 cell-cycle arrest using Chinese hamster ovary (CHO) cells. T-fimbrin, an actin-binding protein, was overexpressed in CHO cells in which G2 arrest had been induced by X-radiation. Northern blot analysis revealed that T-fimbrin gene expression was induced not only by X-radiation but also by a topoisomerase II inhibitor, etoposide. Transfection of CHO cells with a vector encoding T-fimbrin antisense RNA demonstrated that reduced T-fimbrin expression induced alterations in cell-cycle control; radiation-induced G2 arrest was short and decreased in cells transfected with antisense T-fimbrin. Additionally, T-fimbrin gene expression was suppressed in a human colorectal cancer cell line, SW948, because of promoter-specific DNA methylation. These results suggest that downregulation of T-fimbrin may be involved in cancer development through G2/M cell-cycle control in mammalian cells.

MeSH terms

  • Animals
  • Blotting, Northern
  • CHO Cells / metabolism
  • Cell Cycle / radiation effects*
  • Colorectal Neoplasms / genetics*
  • Colorectal Neoplasms / metabolism
  • CpG Islands / genetics*
  • Cricetinae
  • DNA Damage
  • DNA Methylation*
  • DNA Primers / chemistry
  • DNA, Complementary / genetics
  • G2 Phase / radiation effects
  • Gene Expression
  • Gene Expression Profiling / methods
  • Membrane Glycoproteins / genetics*
  • Microfilament Proteins*
  • Promoter Regions, Genetic
  • RNA, Messenger / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Transfection
  • Tumor Cells, Cultured

Substances

  • DNA Primers
  • DNA, Complementary
  • Membrane Glycoproteins
  • Microfilament Proteins
  • RNA, Messenger
  • plastin