Construction Of The Breast Cancer Specific Peptide-mediated HSV-TK/GCV Suicide Gene System And Study On Its Targeted Effection In Vitro

[Abstract] Objective:To investigate the special killing effect of HSV-TK/GCV system on human breast cancer cells in vitro mediated by the breast cancer specific peptide. Methods:PI-TK gene was inserted into the plasmid pET-28a (+)-PI-TK and the recombinant plasmid pET-28a (+)-PI-TK was constructed, After the induction by IPTG,we got the fusion protein which purified by His-Tag,The fusion protein has been identified by the SDS-PAGE condensate gel electrophoresis and Western blotting,The different doses of fusion protein were cultured with MDA-MB-231 cells,the cells morphology was observed by fluorescencete microscopey,CCK-8 method was used to determine the killing effect of 10mg/L GCV on them. Results:Recombinant plasmid pET-28a (+)-PI-TK was constructed successfully,The purified PI-TK fusion protein was confirmed by the SDS-PAGE and Western blotting. CCK-8 assay showed that:Only PI-TK fusion protein does not affect the form and proliferation of the MDA-MB-231 cells, but PI-TK fusion protein with GCV can inhibit targetedly the proliferation of MDA-MB-231 cell in a dose-dependent way, Inhibition ratio of 200μg/ml PI-TK with 10 mg/L GCV come to 68.9±7.57%, IC50 is 152.64μg/ml, without influence to MDA-MB-435 cells (P <0.05). Conclusion:The breast cancer specific peptide-mediated HSV-TK Anti-Tumor system can targetedly kill MDA-MB-231 cells.