LI-JIUAN SHENWu, Fe-Lin LinFe-Lin LinWuFE-LIN LIN WUYeh, Tzyy-HarnTzyy-HarnYehChen, Ying-LuenYing-LuenChenChiu, Yu-ChinYu-ChinChiuCheng, Ju-ChenJu-ChenChengWei, Ming-FengMing-FengWeiShen, Li-JiuanLi-JiuanShen2018-09-102018-09-102014http://www.scopus.com/inward/record.url?eid=2-s2.0-84905459957&partnerID=MN8TOARShttp://scholars.lib.ntu.edu.tw/handle/123456789/383363Recombinant arginine deiminase (rADI) has been used in clinical trials for arginine-auxotrophic cancers. However, the emergence of rADI resistance, due to the overexpression of argininosuccinate synthetase (AS), has introduced an obstacle in its clinical application. Here, we have proposed a strategy for the intracellular delivery of rADI, which depletes both extracellular and intracellular arginine, to restore the sensitivity of rADI-resistant cancer cells. In this study, the C terminus of heparin-binding hemagglutinin adhesion protein from Mycobacterium tuberculosis (HBHAc), which contains 23 amino acids, was used to deliver rADI into rADI-resistant human breast adenocarcinoma cells (MCF-7). Chemical conjugates (l- and d-HBHAc-SPDP-rADI) and a recombinant fusion protein (rHBHAc-ADI) were produced. l- and d-HBHAc-SPDP-rADI showed a significantly higher cellular uptake of rADI by MCF-7 cells compared to that of rADI alone. Cell viability was significantly decreased in a dose-dependent manner in response to l- and d-HBHAc-SPDP-rADI treatments. In addition, the ratio of intracellular concentration of citrulline to arginine in cells treated with l- and d-HBHAc-SPDP-rADI was significantly increased by 1.4- and 1.7-fold, respectively, compared with that obtained in cells treated with rADI alone (p < 0.001). Similar results were obtained with the recombinant fusion protein rHBHAc-ADI. Our study demonstrates that the increased cellular uptake of rADI by HBHAc modification can restore the sensitivity of rADI treatment in MCF-7 cells. rHBHAc-ADI may represent a novel class of antitumor enzyme with an intracellular mechanism that is independent of AS expression. ? 2014 American Chemical Society.cancer; HBHA; heparin-binding hemagglutinin adhesion; intracellular delivery; rADI; recombinant arginine deiminase; resistance[SDGs]SDG3amino acid; arginine; arginine deiminase; citrulline; hemagglutinin; heparin binding hemagglutinin adhesion peptide; heparin binding protein; hybrid protein; unclassified drug; arginine deiminase; argininosuccinate synthase; citrulline; fluorescein; heparin-binding hemagglutinin; hybrid protein; hydrolase; lectin; peptide; article; breast adenocarcinoma; cancer cell; cancer resistance; cell viability; concentration (parameters); conjugate; controlled study; human; human cell; MCF 7 cell line; Mycobacterium tuberculosis; priority journal; synthesis; cell survival; chemistry; cytoplasm; dose response; drug delivery system; drug effects; drug resistance; endocytosis; metabolism; Neoplasms; tumor cell line; Amino Acids; Argininosuccinate Synthase; Cell Line, Tumor; Cell Survival; Citrulline; Cytoplasm; Dose-Response Relationship, Drug; Drug Delivery Systems; Drug Resistance, Neoplasm; Endocytosis; Fluorescein; Humans; Hydrolases; Lectins; MCF-7 Cells; Mycobacterium tuberculosis; Neoplasms; Peptides; Recombinant Fusion Proteinsjournal article10.1021/mp5001372