title: Unravelling the potential of a new uracil phosphoribosyltransferase (UPRT) from Arabidopsis thaliana in sensitizing HeLa cells towards 5-fluorouracil.
abstract: In silico studies with uracil phosphoribosyltransferase from Arabidopsis thaliana (AtUPRT) revealed its lower binding energies for uracil and 5-fluorouracil (5-FU) as compared to those of bacterial UPRT indicating the prospective of AtUPRT in gene therapy implications. Hence, AtUPRT was cloned and stably expressed in cervical cancer cells (HeLa) to investigate the effect of prodrug 5-FU on these transfected cancer cells. The treatment of AtUPRT-expressing HeLa (HeLa-UPP) cells with 5-FU for 72h resulted in significant decrease in cell viability. Moreover, 5-FU was observed to induce apoptosis and perturb mitochondrial membrane potential in HeLa-UPP cells. While cell cycle analysis revealed significant S-phase arrest as a result of 5-FU treatment in HeLa-UPP cells, quantitative gene expression analysis demonstrated simultaneous upregulation of important cell cycle related genes, cyclin D1 and p21. The survival fractions of non-transfected, vector-transfected and AtUPRT-transfected HeLa cells, following 5-FU treatment, were calculated to be 0.425, 0.366 and 0.227, respectively.
authors.affiliation: Department of Biosciences & Bioengineering, Indian Institute of Technology Guwahati, Guwahati-39, Assam, India.
authors.forename: Sharmila
authors.initials: S
authors.lastname: Narayanan
authors.affiliation: Centre for Nanotechnology, Indian Institute of Technology Guwahati, Guwahati-39, Assam, India.
authors.forename: Pallab
authors.initials: P
authors.lastname: Sanpui
authors.affiliation: Department of Biosciences & Bioengineering, Indian Institute of Technology Guwahati, Guwahati-39, Assam, India.
authors.forename: Lingaraj
authors.initials: L
authors.lastname: Sahoo
authors.affiliation: Department of Biosciences & Bioengineering, Indian Institute of Technology Guwahati, Guwahati-39, Assam, India; Centre for Nanotechnology, Indian Institute of Technology Guwahati, Guwahati-39, Assam, India. Electronic address: sghosh@iitg.ernet.in.
authors.forename: Siddhartha Sankar
authors.initials: SS
authors.lastname: Ghosh
mesh_terms: D017360:Arabidopsis / Q000201:enzymology / Q000235:genetics*; D029681:Arabidopsis Proteins / Q000096:biosynthesis* / Q000235:genetics; D019008:Drug Resistance, Neoplasm / Q000187:drug effects* / Q000235:genetics*; D005472:Fluorouracil / Q000494:pharmacology*; D006367:HeLa Cells; D006801:Humans; D010430:Pentosyltransferases / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D029681:Arabidopsis Proteins; D010430:Pentosyltransferases; C023269:uracil phosphoribosyltransferase; D005472:Fluorouracil
keywords: 5-fluorouracil; Apoptosis; Arabidopsis thaliana; Cell cycle; Gene therapy; Uracil phosphoribosyltransferase