文献类型：期刊文献

英文题名：Enhanced anti-tumor activity of trichosanthin after combination with a human-derived cell-penetrating peptide, and a possible mechanism of activity

作者：Lu, Ye-Zhou[1];Li, Peng Fei[1];Li, Yan-Zhong[1];Luo, Fan[1];Guo, Chao[1];Lin, Bin[1];Cao, Xue-Wei[1];Zhao, Jian[1];Wang, Fu-Jun[2,3]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China;[2]Shanghai Univ Tradit Chinese Med, Inst Chinese Mat Med, 1200 Cailun Rd, Shanghai 201203, Peoples R China;[3]Zhejiang Reachall Pharmaceut Co Ltd, Dongyang 322100, Zhejiang, Peoples R China

年份：2016

卷号：112

起止页码：183

外文期刊名：FITOTERAPIA

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000380865400023)】；

基金：This work was sponsored by the National Natural Science Foundation of China (grant no. 81571795).

语种：英文

外文关键词：Trichosanthin; Cell-penetrating peptide; Tumor therapy; Apoptosis

摘要：Trichosanthin (TCS), a type I ribosome-inactivating protein (RIP-I) and renowned Chinese traditional medicine, displays a broad spectrum of biological and pharmacological properties. Particularly, its anti-tumor activity has received a great deal of attention. However, the cellular mechanism for TCS uptake varies with different tumor cell lines, leading to discrepancies in its reported ability to penetrate cells. In this study, HBD, a human derived cell-penetrating peptide (CPP), was used to improve the delivery of TCS into several types of tumor cells, including HeLa cells. Recombinant TCS (rTCS) with or without the fused HBD peptide was expressed in Escherichia coli cells and successfully purified by Ni-NTA affinity chromatography. The cellular uptake efficiency of FITC-labelled-rTCS-HBD was observed in HeLa cells and compared with the uptake efficiency of non-HBD conjugated rTCS under the same conditions using laser confocal microscopy. Moreover, the IC50 value of rTCS-HBD in the tested tumor cells was much lower than that of rTCS, indicating that HBD could efficiently deliver the rTCS into tumor cells. When compared with rTCS, rTCS-HBD induced higher rates of apoptosis in HeLa cells as analyzed by flow cytometry. Furthermore, the apoptotic events observed in HeLa cells incubated with HBD-fused rTCS included activation of Caspase-9, decrease in the Bcl-2/Bax ratio, and cleavage of PARP. These results strongly suggest the participation of mitochondria in apoptosis. This report illustrates one possible method for achieving the efficient transport of TCS into cells using a CPP as a vector, and increases the likelihood that TCS can be used in the clinic. (C) 2016 Elsevier B.V. All rights reserved.