Synthetic radiation-inducible promoter mediated CDglyTK gene in treatment of Tca8113 cells
Yu, D.-S.; Huang, H.-Z.; Xie, Q.; Wang, A.-X.; Hu, X.-W.; Liu, X.-q.
Hua Xi Kou Qiang Yi Xue Za Zhi 24(3): 269-272
2006
ISSN/ISBN: 1000-1182 PMID: 16845968 Document Number: 600387
To observe the therapeutic effect of CDglyTK gene mediated by synthetic radiation-inducible promoter in the treatment of Tca8113 cells. CDglyTK gene in pCEA-CDglyTK was subcloned into pcDNA3.1 (+) to construct plasmid pcDNA3.1 (+)-CDglyTK, and then the synthetic radiation-inducible promoter in pMD18 -T -E was inserted into pcDNA3.1 (+) -CDglyTK to construct plasmid pcDNA3.1 (+ )/E -CDglyTK. The recombinant plasmid was transfected into Tca8113 cells by lipofectamine, and then exposed to 3 Gy irradiation. Cytotoxicity was evaluated by MTT. The expression of CDglyTK gene was detected by RT-PCR. The apoptosis and proliferation were examined by flow cytomtery. The plasmid pcDNA3.1 (+)/E-CDglyTK was constructed successfully. The comparative survival rate of Tca8113 cells was markedly decreased by induction irradiation. Up-regulation of CDglyTK expression was found in Tca8113 cells exposed to irradiation. The apoptosis index (AI) of Tca8113 cells exposed to irradiation was higher than that of Tca8113 cells without irradiation, the other way round, the proliferation index (PI) of Tca8113 cells exposed to irradiation was lower than that of Tca8113 cells without irradiation. The synthetic radiation-inducible promoter can be served as a molecular switch to improve the expression of CDglyTK gene in Tca8113 cells, and low dose induction radiation can significantly improve the therapeutic efficiency.