title: TAT Modification of Alpha-Helical Anticancer Peptides to Improve Specificity and Efficacy.
abstract: HPRP-A1 is an amphipathic α-helical anticancer peptide (ACP) derived from the N-terminus of ribosomal protein L1 (RpL1) of Helicobacter pylori. In our previously study, HPRP-A1 has been reported that induced HeLa cell apoptosis in a caspase-dependent approach and involved both by the death receptor 'extrinsic' pathway and the mitochondria 'intrinsic' pathway. Here we report the construction of a new hybrid peptide, HPRP-A1-TAT, comprising the cell-permeating peptide TAT linked to the C-terminus of HPRP-A1. This peptide exhibits higher anticancer activity against HeLa cells with lower toxicity against human RBC than HPRP-A1. Two FITC-labeled peptides, FITC-HPRP-A1 and FITC-HPRP-A1-TAT, were used to investigate and compare the cellular uptake mechanism using fluorescence spectra and flow cytometry. Compared with HPRP-A1, HPRP-A1-TAT quickly crossed cell, entered the cytoplasm via endocytosis, and disrupted the cell membrane integrity. HPRP-A1-TAT exhibited stronger anticancer activity than HPRP-A1 at the same concentration by increasing early apoptosis of HeLa cells and inducing caspase activity. Notably, after 24 h, the cellular concentration of HPRP-A1-TAT was higher than that of HPRP-A1. This result suggests that TAT protects HPRP-A1 against degradation, likely due to its high number of positively charged amino acids or the further release of peptides into cancer cells from endocytotic vesicles. We believe that this TAT modification approach may provide an effective new strategy for improving the therapeutic index and anticancer activity of ACPs for clinical use.
authors.affiliation: Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education, Jilin University, Changchun, China; School of Life Sciences, Jilin University, Changchun, China.
authors.forename: Xueyu
authors.initials: X
authors.lastname: Hao
authors.affiliation: Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education, Jilin University, Changchun, China; School of Life Sciences, Jilin University, Changchun, China.
authors.forename: Qiuyan
authors.initials: Q
authors.lastname: Yan
authors.affiliation: Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education, Jilin University, Changchun, China; School of Life Sciences, Jilin University, Changchun, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Zhao
authors.affiliation: Changchun ProteLight Pharmaceutical & Biotechnology Co., Ltd., Changchun, China.
authors.forename: Wenren
authors.initials: W
authors.lastname: Wang
authors.affiliation: Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education, Jilin University, Changchun, China; School of Life Sciences, Jilin University, Changchun, China; National Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun, China.
authors.forename: Yibing
authors.initials: Y
authors.lastname: Huang
authors.affiliation: Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education, Jilin University, Changchun, China; School of Life Sciences, Jilin University, Changchun, China; National Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun, China.
authors.forename: Yuxin
authors.initials: Y
authors.lastname: Chen
mesh_terms: D000970:Antineoplastic Agents / Q000138:chemical synthesis* / Q000737:chemistry / Q000493:pharmacokinetics* / Q000494:pharmacology; D017209:Apoptosis; D020169:Caspases / Q000378:metabolism; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006367:HeLa Cells; D056945:Hep G2 Cells; D006801:Humans; D009928:Organ Specificity; D010446:Peptide Fragments / Q000138:chemical synthesis* / Q000737:chemistry / Q000493:pharmacokinetics* / Q000494:pharmacology; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000737:chemistry*
chemical_list: D000970:Antineoplastic Agents; D010446:Peptide Fragments; D054322:tat Gene Products, Human Immunodeficiency Virus; C490722:tat peptide (49-57), Human immunodeficiency virus 1; D020169:Caspases