Enhancement of thymidine phosphorylase expression in the human colorectal cancer cell lines by transfected TP cDNA with lentiviral vector
WANG Qi-Wen
LIU Jian
YE Dian-Jun
ZHANG Ji-min
Abstract：Objective To build the human colorectal cancer cell lines with the high expression of thymidine phos -phorylase (TP) by transfected TP cDNA with lentiviral vector .Methods Human colorectal cancer cell lines SW480, LOVO, HT29 and LS174T were transfected with human lentiviral vector TP cDNA by pLenti 6.3_MCS_IRES2 -EGFP. The transfection efficiency was analyzed by flow cytometer , the mRNA expression of TP was detected by RT -PCR and the TP level was detected by Western blot .Results The stabilized transfection efficiency was about 95% past 5 generations. Comparing with parental SW480, LOVO, HT29 and LS174T, the mRNA expression of SW480 -TP, LOVO -TP, HT29 -TP and LS174T -TP were significantly up -regulated by (694.56 ±171.53), (282.46 ±86.85), (8.45 ±0.15) and (2 615.02 ±253.97) folds, respectively (P <0.05).The Western blot showed that the TP levels of SW 480 -TP, LOVO -TP, HT29 -TP and LS174T -TP were obvious up -regulated.Conclusion Stabilized transfections of 4 cell lines with a high TP expression is successfully constructed by lentiviral vector , with significantly up -regulation of TP in mRNA and protein levels.
Keywords：thymidine phosphorylasegene transfectionlentiviral vectorSW480LOVOHT29LS174T
Publication Date：2015-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：5( 30-34 )