Cloning and expression of human uridine phosphorylase

Biochem Biophys Res Commun. 1995 Nov 2;216(1):265-72. doi: 10.1006/bbrc.1995.2619.

Abstract

Using a mouse cDNA probe we have identified a human uridine phosphorylase cDNA clone from the cDNA library of a human colorectal tumor cell line, HCT116. The recombinant human uridine phosphorylase expressed in COS-7 cells demonstrated specific enzyme activity with uridine as the substrate; this activity was inhibited by the competitive inhibitor 2,2'-anhydro-5-ethyluridine. Northern blot analysis with the cDNA as a probe demonstrated high levels of mRNA expression in several tumor cell lines but very low level in normal cell, WI-38. The expression of uridine phosphorylase mRNA in HCT-116 cells was further enhanced by treating the cells with vitamin D3 and the inflammatory cytokines: tumor necrosis factor alpha, interleukin 1 alpha and interferon gamma.

Publication types

  • Comparative Study

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Calcitriol / pharmacology
  • Cell Line
  • Chlorocebus aethiops
  • Cloning, Molecular
  • Colorectal Neoplasms
  • Cytokines / pharmacology
  • DNA Probes
  • DNA, Complementary
  • Gene Expression* / drug effects
  • Humans
  • Interferon-gamma / pharmacology
  • Interleukin-1 / pharmacology
  • Kinetics
  • Mice
  • Molecular Sequence Data
  • RNA, Messenger / biosynthesis
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism
  • Reference Values
  • Sequence Homology, Amino Acid
  • Tumor Cells, Cultured
  • Tumor Necrosis Factor-alpha / pharmacology
  • Uridine Phosphorylase / biosynthesis*
  • Uridine Phosphorylase / chemistry
  • Uridine Phosphorylase / metabolism

Substances

  • Cytokines
  • DNA Probes
  • DNA, Complementary
  • Interleukin-1
  • RNA, Messenger
  • Recombinant Proteins
  • Tumor Necrosis Factor-alpha
  • Interferon-gamma
  • Uridine Phosphorylase
  • Calcitriol

Associated data

  • GENBANK/X90858