Transfection of thymidine phosphorylase cDNA with lentiviral vector enhances the anticancer effect of 5'-deoxy-5-fluorouridine on colorectal cancer cell lines HT29 and LS174T

• Published in: Zhōnghuá zhŏngliú zázhì Vol. 37; no. 1; p. 18
• Main Authors: Ye, Dianjun, Wang, Qiwen, Zhang, Jimin, Liu, Qi
• Format: Journal Article
• Language: Chinese
• Published: China 01.01.2015
• Subjects: Antineoplastic Agents - pharmacology; Cell Line; Cell Line, Tumor; Colorectal Neoplasms - genetics; Colorectal Neoplasms - metabolism; DNA, Complementary; Floxuridine - pharmacology; Fluorouracil; Genetic Vectors; HT29 Cells; Humans; RNA, Messenger; Thymidine Phosphorylase - genetics; Thymidine Phosphorylase - metabolism; Transfection
• ISSN: 0253-3766
• DOI: 10.3760/cma.j.issn.0253-3766.2015.01.004

To explore the changes of anticancer efficiency of 5'-deoxy-5-fluorouridine (5'-DFUR) and 5-fluorouracil (5-Fu) in colorectal cancer cell line HT29 and LS174T cells after transfection of thymidine phosphorylase (TP) cDNA with a lentiviral vector. TP cDNA was transfected into human colorectal cancer cell lines HT29 and LS174T with the lentiviral vector pLenti6.3-MCS-IRES2-EGFP, and the transfection efficiency of the two cell lines passed 5 generations was analyzed by flow cytometry. The expression of TP protein and the relative quantitative expression of TP mRNA in these 2 cell lines were detected by Western blot and RT-PCR, respectively. The 50% inhibitory concentration (IC50) of 5'-DFUR and 5-Fu in both HT29 and LS174T parent cells and TP-transfected cells were assessed by MTS assay. Finally, the concentration of converted 5-Fu was detected by high performance liquid chromatography (HPLC) either in the medium containing a series of concentrations of 5'-DFUR, in which HT29/HT29-TP or LS174T/LS174T-TP cells we