文献类型：期刊文献

中文题名：pH敏感型聚合物胶束对I型志贺毒素A亚基的递送及细胞毒性

英文题名：Delivery and Cytotoxicity of Type I Shiga Toxin A Subunit by pH-Sensitive Polymer Micelles

作者：孙敏佳[1,2];刘晔宏[1];薛依桐[2];徐俊[1];王彤[1];徐首红[1];张俊琪[2]

机构：[1]华东理工大学化学与分子工程学院,结构可控先进功能材料及其制备教育部重点实验室,上海200237;[2]复旦大学基础医学院教育部/卫建委医学分子病毒学重点实验室&病原生物系,上海200032

年份：2022

卷号：48

期号：2

起止页码：213

中文期刊名：华东理工大学学报（自然科学版）

外文期刊名：Journal of East China University of Science and Technology

收录：Scopus;北大核心:【北大核心2020】；CSCD:【CSCD_E2021_2022】；

基金：国家自然科学基金(31400124,21776071,22078087);国家科技重大专项(2018ZX10101003-005-010)。

语种：中文

中文关键词：I型志贺毒素;pH敏感型聚合物胶束;蛋白类药物;胞内递送;肿瘤治疗

外文关键词：Shiga toxin type I;pH sensitive polymer micelles;peptide drugs;targeted delivery;tumor therapy

摘要：探究了pH敏感型聚合物胶束递送和释放I型志贺毒素A亚基至肿瘤细胞Hela的效率和功能。首先将在大肠杆菌中分别重组表达I型志贺毒素A亚基Stx1A和减毒突变A亚基Mu-Stx1A,然后将对pH敏感的聚合物胶束PEG_(8)-PDPA_(100)-PEG_(8)(其中PEG为聚乙二醇,PDPA为聚异丙基甲基烯酸酯)分别运载至宫颈癌细胞Hela中。体外活性实验证明重组蛋白Stx1A具有明显的抑制蛋白合成作用,而减毒变异型Mu-Stx1A不具备这种作用。用聚合物胶束将Stx1A及Mu-Stx1A转运至Hela细胞中,发现随着蛋白浓度的增加细胞转染效率增大。包载了Stx1A的胶束进入细胞后,释放活性Stx1A导致细胞病变和凋亡,该现象随着Stx1A浓度的增加表现更显著。实验证明聚合物胶束可以成功包载、运输和稳定释放具有活性的Stx1A分子至肿瘤细胞内,发挥毒性功能诱导细胞程序性死亡。该聚合物胶束可以在蛋白类药物运输中发挥有效作用,为后续I型志贺毒素A亚基在肿瘤治疗中的应用性研究提供重要的理论基础。
This paper aims to explore the efficiency and function of pH sensitive polymer micelles for targeted delivery and release of Shiga toxin 1 subunit A to HeLa cells.Recombinantly expressed the Shiga toxin 1 A subunit(Stx1A)and the attenuated variant A subunit(Mu-Stx1A)in Escherichia coli,and used pH-sensitive polymer micelles formed by self-assembly of PEG_(8)-PDPA_(100)-PEG_(8) to deliver them to the cervical cancer cells Hela.In vitro activity test showed that the recombinant protein Stx1A could significantly inhibit protein synthesis,but the attenuated mutant Mu-Stx1A could not.Stx1A and Mu-Stx1A were successfully transported into HeLa cells by the polymeric micelles,and the transfection efficiency increased with the protein concentration.After Stx1A micelles entered the cells,it successfully released the active subunit Stx1A,leading to cytopathy and apoptosis,which became more evident with the concentration of Stx1A.Experiments have proved that the polymer micelles can successfully encapsulate,transport and stably release Stx1A molecules into tumor cells,exerting toxic functions to induce programmed cell death.This work indicates that polymer micelles can play an effective role in protein transport,which provides a theoretical basis for the subsequent research and application of Shiga toxin 1 subunit A in tumor therapy.