Activation of c-myc gene mediates the mitogenic effects of oxalate in LLC-PK1 cells, a line of renal epithelial cells

Kidney Int. 1996 Nov;50(5):1525-30. doi: 10.1038/ki.1996.467.

Abstract

Recent studies on LLC-PK1 cells demonstrated that oxalate, a simple dicarboxylic acid, acts as a mitogen for these renal epithelial cells. Exposure to oxalate initiates DNA synthesis, induces the expression of one of the early growth response genes c-myc and stimulates proliferation of quiescent cultures of LLC-PK1 cells. The present studies examined the possibility that expression of the c-myc protooncogene is obligatory for this mitogenic response. Specifically we determined whether pretreatment with c-myc antisense oligonucleotides would block the proliferative effects of oxalate in LLC-PK1 cells. Quiescent cultures of LLC-PK1 cells were exposed to oxalate in the presence and absence of c-myc antisense and the effects of oxalate on c-myc protein expression (Myc), DNA synthesis and cell growth were assessed. Exposure of cells to oxalate alone increased the expression of Myc within two hours. Pretreatment with c-myc antisense abolished this response. Further, pretreatment of cells with c-myc antisense but not nonsense oligonucleotides blocked the oxalate-induced initiation of DNA synthesis. Increases in cell number in response to oxalate (measured after 72 hr exposure) were also blocked by exposure to c-myc antisense. These findings suggest that c-myc gene expression is critical for the mitogenic effects of oxalate in LLC-PK1 cells.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Blotting, Northern
  • Cell Count
  • Culture Media, Serum-Free
  • Epithelial Cells
  • Epithelium / drug effects
  • Epithelium / metabolism
  • Genes, myc / physiology*
  • Kidney / cytology
  • Kidney / drug effects
  • Kidney / metabolism*
  • LLC-PK1 Cells
  • Mitogens / pharmacology*
  • Oligonucleotides, Antisense / pharmacology
  • Oxalates / pharmacology*
  • Oxalic Acid
  • Proto-Oncogene Proteins c-myc / biosynthesis*
  • Proto-Oncogene Proteins c-myc / genetics
  • Swine
  • Thymidine / metabolism

Substances

  • Culture Media, Serum-Free
  • Mitogens
  • Oligonucleotides, Antisense
  • Oxalates
  • Proto-Oncogene Proteins c-myc
  • Oxalic Acid
  • Thymidine