天花粉蛋白的抗肿瘤药效研究

AbstractTrichosanthin (TCS),a type-I ribosome-inactivating protein (RIP),is extracted from the root tuberof the Chinese medicinal plant Trichosanthes kirilowii.TCS is reported to be a potential therapeuticagent for possessing a variety of biological activities,including anti-tumor activities and anti-HIVinfection.TCS is highly toxic to melanoma cells and choriocarcinoma cells,but has only slight effectson fibroblasts and hepatoma cells,which limits its anti-tumor applications.Cell-penetrating peptides(CPPs)are short peptides,which are rich in basic amino acid,with a remarkable capacity for membranetranslocation.The present study uses a human derived cell-penetrating peptide fused with TCSexpression to carry it into tumor cells effectively.The present study elaborates the fusion expression and pharmacological properties of this novelanti-cancer drug derived from TCS.The fusion protein of recombinant TCS is highly expressed inprokaryotic E.coli and purified with nickel column affinity chromatography.The study analyzed thefusion protein with MTT,confocal laser scanning microscope,flow cytometry.Theresults suggest that TCS-HBP shows better inhibition for tumor cells in vitro and a broader spectrum oftumor cells compared to TCS.The results of Western Blot suggest that the TCS-HBP may induceapoptosis of Hela cells through mitochondrial pathway.The present study is membrane translocationoriented,which fused the human derived cell-penetrating peptide with Trichosanthin.We elaborate themembrane translocation and the apoptosis pathway of the fusion protein TCS-HBP,hoping that it couldprovide some useful reference for the delivery and higher bioactivity of TCS.Keywords:Trichosanthin,Cell-penetrating peptides,Apoptosis,Drug delivery之