Cloning and expression of human uridine phosphorylase.
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.
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002117:Calcitriol / Q000494:pharmacology; D002460:Cell Line; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D015179:Colorectal Neoplasms; D016207:Cytokines / Q000494:pharmacology; D015342:DNA Probes; D018076:DNA, Complementary; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology; D007375:Interleukin-1 / Q000494:pharmacology; D007700:Kinetics; D051379:Mice; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D012016:Reference Values; D017386:Sequence Homology, Amino Acid; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D014543:Uridine Phosphorylase / Q000096:biosynthesis* / Q000737:chemistry / Q000378:metabolism
D016207:Cytokines; D015342:DNA Probes; D018076:DNA, Complementary; D007375:Interleukin-1; D012333:RNA, Messenger; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha; D007371:Interferon-gamma; D014543:Uridine Phosphorylase; D002117:Calcitriol