基于开环反应构建新型还原响应型支化聚赖氨酸基因载体

陈艺铭; 祁宇; 胡文婷; 郑迪; 俞丙然; 徐福建

doi:10.11777/j.issn1000-3304.2019.18260

您当前的位置：

首页 >

文章列表页 >

基于开环反应构建新型还原响应型支化聚赖氨酸基因载体

研究论文 | 更新时间：2021-01-26

- 基于开环反应构建新型还原响应型支化聚赖氨酸基因载体
- Novel Reduction-responsive Branched Polylysine via Ring-opening Reactions for Gene Delivery
- 高分子学报 2019年50卷第6期页码：602-612
- 作者机构：
  
  生物医用材料北京实验室北京化工大学　北京 100029
- 作者简介：
  
  E-mail: yubr@mail.buct.edu.cn
  E-mail: xufj@mail.buct.edu.cn
- 基金信息：
  
  国家自然科学基金(基金号 51733001，31771045和21875014)和中央高校基本业务费(项目号 PYBZ1808，PYBZ1814)资助项目()
- DOI：10.11777/j.issn1000-3304.2019.18260
  中图分类号：
- 纸质出版日期：2019-6，
  
  网络出版日期：2019-1-31，
  
  收稿日期：2018-11-29，
  
  修回日期：2018-12-22，
扫描看全文
陈艺铭, 祁宇, 胡文婷, 郑迪, 俞丙然, 徐福建. 基于开环反应构建新型还原响应型支化聚赖氨酸基因载体[J]. 高分子学报, 2019,50(6):602-612.

Yi-ming Chen, Yu Qi, Wen-ting Hu, Di Zheng, Bing-ran Yu, Fu-jian Xu. Novel Reduction-responsive Branched Polylysine via Ring-opening Reactions for Gene Delivery[J]. Acta Polymerica Sinica, 2019,50(6):602-612.
陈艺铭, 祁宇, 胡文婷, 郑迪, 俞丙然, 徐福建. 基于开环反应构建新型还原响应型支化聚赖氨酸基因载体[J]. 高分子学报, 2019,50(6):602-612. DOI： 10.11777/j.issn1000-3304.2019.18260.

Yi-ming Chen, Yu Qi, Wen-ting Hu, Di Zheng, Bing-ran Yu, Fu-jian Xu. Novel Reduction-responsive Branched Polylysine via Ring-opening Reactions for Gene Delivery[J]. Acta Polymerica Sinica, 2019,50(6):602-612. DOI： 10.11777/j.issn1000-3304.2019.18260.

摘要

通过“一锅法”开环反应构建了一种含有双硫键的还原响应型可降解支化聚赖氨酸基因载体(SS-HP). 同时通过相似的方法合成了不含双硫键的支化聚赖氨酸基因载体(CC-HP)作为对照. 聚阳离子/pDNA复合物的水合粒径和电位通过动态光散射仪测定，其在还原性环境中的降解情况通过凝胶阻滞电泳、原子力显微镜测试评价. SS-HP和CC-HP的体外转染能力及细胞毒性通过荧光素酶报告基因和MTT法在脑胶质瘤C6和肝癌HepG2细胞中测定. 癌细胞中过量表达的谷胱甘肽能够使SS-HP在细胞内降解，加速pDNA的释放，使该载体有良好的基因转染能力. 同时，载体的可降解性以及由开环反应给载体引入的大量羟基，使载体有较低的细胞毒性. KillerRed蛋白是一种在可见光照射下能产生单线态氧的红色荧光蛋白. 使用SS-HP携载pKillerRed (pKR)质粒进行体外光动力抗癌评价. SS-HP/pKR转染C6细胞后能成功表达出KillerRed蛋白，光照后可以促进肿瘤细胞的凋亡.

Abstract

Gene therapy is widely concerned as an excellent treatment for cancer. One of the most important things in gene therapy is to construct gene delivery systems with good biodegradability

biocompatibility

and gene delivery capability. Biodegradable non-viral gene vectors based on different microenvironments between cancer cells and normal cells have been paid more attention. In this work

the reduction-responsive branched polylysine (SS-HP) with disulfide bonds was synthesized

via

a one-pot ring-opening reaction. To make a comparison

the branched polylysine without disulfide bonds (CC-HP) was synthesized by the same method. All the SS-HP/pDNA and CC-HP/pDNA complexes with various weight ratios were prepared by mixing polycation-based solution and pDNA solution completely

and stood for 30 min. The particle size and zeta potential of SS-HP/pDNA and CC-HP/pDNA were measured by dynamic light scattering (DLS). The degradability of polyplexes in reductive environment was visualized by agarose gel electrophoresis and atomic force microscopy (AFM). The

in vitro

transfection efficiencies and cell viability of SS-HP and CC-HP were evaluated in C6 and HepG2 cell lines using luciferase reporter gene

green fluorescence protein gene

and MTT assay. The concentration of reductive glutathione (GSH) is higher in some cancer cells than that in normal cells. SS-HP showed high gene transfection efficiency

in vitro

due to the breakdown of reduction-responsive disulfide bonds. Moreover

SS-HP exhibited low cytotoxicity due to the good biodegradability of SS-HP and plenty of hydroxy groups induced by ring-opening reactions. KillerRed protein is a red fluorescence protein which could produce reactive oxygen species (ROS) upon the induction of visible light. From

in vitro

antitumor assays

the plasmid pKillerRed (pKR) delivered by SS-HP was expressed in C6 cells. KillerRed protein expressed in C6 cells could contribute to the cell apoptosis

via

photodynamic therapy (PDT). This study provides a novel approach for designing the next-generation gene delivery systems.

关键词

还原响应生物可降解支化聚合物基因载体

Keywords

Reduction-responsiveBiodegradableBranched polymerGene delivery

references

Fan W, Yung B, Huang P, Chen X . Chem Rev , 2017 . 117 ( 22 ): 13566 - 13638 . DOI:10.1021/acs.chemrev.7b00258http://doi.org/10.1021/acs.chemrev.7b00258 .

Mahalingam S M, Kularatne S A, Myers C H, Gagare P, Norshi M, Liu X, Singhal S, Low P S . J Med Chem , 2018 . 61 ( 21 ): 9637 - 9646 . DOI:10.1021/acs.jmedchem.8b01115http://doi.org/10.1021/acs.jmedchem.8b01115 .

Chen J, Tian H, Dong X, Guo Z, Jiao Z, Li F, Kano A, Maruyama A, Chen X . Macromol Biosci , 2013 . 13 ( 10 ): 1438 - 1446 . DOI:10.1002/mabi.201300211http://doi.org/10.1002/mabi.201300211 .

Guan X, Li Y, Jiao Z, Lin L, Chen J, Guo Z, Tian H, Chen X . ACS Appl Mater Interfaces , 2015 . 7 ( 5 ): 3207 - 3215 . DOI:10.1021/am5078123http://doi.org/10.1021/am5078123 .

Lin Lin(林琳), Guo Zhaopei(郭兆培), Chen Jie(陈杰), Tian Huayu(田华雨), Chen Xuesi(陈学思) . 高分子学报 , Acta Polymerica Sinica , 2017 . ( 2 ): 321 - 328.

Wang Y, Xiao H, Fang J, Yu X, Su Z, Cheng D, Shuai X . Chem Commun , 2016 . 52 ( 6 ): 1194 - 1197 . DOI:10.1039/C5CC09181Khttp://doi.org/10.1039/C5CC09181K .

Liu X, Xiang J J, Zhu D C, Jiang L M, Zhou Z X, Tang J B, Liu X R, Huang Y Z, Shen Y Q . Adv Mater , 2016 . 28 ( 9 ): 1743 - 1752 . DOI:10.1002/adma.201504288http://doi.org/10.1002/adma.201504288 .

Putnam D . Nat Mater , 2006 . 5 ( 6 ): 439 - 451 . DOI:10.1038/nmat1645http://doi.org/10.1038/nmat1645 .

Fu Huili(付慧莉), Cheng Sixue(程巳雪), Zhuo Renxi(卓仁禧) . 高分子学报 , Acta Polymerica Sinica , 2009 . ( 2 ): 97 - 103 . DOI:10.3321/j.issn:1000-3304.2009.02.001http://doi.org/10.3321/j.issn:1000-3304.2009.02.001 .

Kim H, Kim W J . Small , 2014 . 10 ( 1 ): 117 - 126 . DOI:10.1002/smll.201202636http://doi.org/10.1002/smll.201202636 .

Yamano S, Dai J, Hanatani S, Haku K, Yamanaka T, Ishioka M, Takayama T, Yuvienco C, Khapli S, Moursi A M, Montclare J K . Biomaterials , 2014 . 35 ( 5 ): 1705 - 1715 . DOI:10.1016/j.biomaterials.2013.11.012http://doi.org/10.1016/j.biomaterials.2013.11.012 .

Qian X, Long L, Shi Z, Liu C, Qiu M, Sheng J, Pu P, Yuan X, Ren Y, Kang C . Biomaterials , 2014 . 35 ( 7 ): 2322 - 2335 . DOI:10.1016/j.biomaterials.2013.11.039http://doi.org/10.1016/j.biomaterials.2013.11.039 .

Li Y, Bing X, Tao B, Liu W . Biomaterials , 2015 . 55 12 - 23 . DOI:10.1016/j.biomaterials.2015.03.034http://doi.org/10.1016/j.biomaterials.2015.03.034 .

Ma D, Lin Q M, Zhang L M, Liang Y Y, Xue W . Biomaterials , 2014 . 35 ( 14 ): 4357 - 4367 . DOI:10.1016/j.biomaterials.2014.01.070http://doi.org/10.1016/j.biomaterials.2014.01.070 .

Zhang Qian(张倩), Yuan Zhi(袁直) . 高分子学报 , Acta Polymerica Sinica , 2018 . ( 7 ): 776 - 785.

Wang D, Zhao T, Zhu X, Yan D, Wang W . Chem Soc Rev , 2015 . 44 ( 12 ): 4023 - 4071 . DOI:10.1039/C4CS00229Fhttp://doi.org/10.1039/C4CS00229F .

Jin H, Huang W, Zhu X, Zhou Y, Yan D . Chem Soc Rev , 2012 . 41 ( 18 ): 5986 - 5997 . DOI:10.1039/c2cs35130ghttp://doi.org/10.1039/c2cs35130g .

Huang Y, Wang D, Zhu X, Yan D, Chen R . Polym Chem , 2015 . 6 ( 15 ): 2794 - 2812 . DOI:10.1039/C5PY00144Ghttp://doi.org/10.1039/C5PY00144G .

Wang K, Peng H, Thurecht K J, Puttick S, Whittaker A K . Polym Chem , 2016 . 7 ( 5 ): 1059 - 1069 . DOI:10.1039/C5PY01707Fhttp://doi.org/10.1039/C5PY01707F .

Huang Y, Ding X, Qi Y, Yu B, Xu F J . Biomaterials , 2016 . 106 134 - 143 . DOI:10.1016/j.biomaterials.2016.08.025http://doi.org/10.1016/j.biomaterials.2016.08.025 .

Fletcher N L, Houston Z H, Simpson J D, Veedu R N, Thurecht K J . Chem Commun , 2018 . 54 ( 82 ): 11538 - 11541 . DOI:10.1039/C8CC05831Hhttp://doi.org/10.1039/C8CC05831H .

Zhang C, Moonshi S S, Wang W, Ta H T, Han Y, Han F Y, Peng H, Kral P, Rolfe B E, Gooding J J, Gaus K, Whittaker A K . ACS Nano , 2018 . 12 ( 9 ): 9162 - 9176 . DOI:10.1021/acsnano.8b03726http://doi.org/10.1021/acsnano.8b03726 .

Zhang S, Liu Y, Derakhshanfar S, He W, Huang Q, Dong S, Rao J, Luo G X, Zhong W, Liao W, Shi M, Xing M . Adv Healthc Mater , 2018 . 1800118 .

Xu H, Cao W, Zhang X . Acc Chem Res , 2013 . 46 ( 7 ): 1647 - 1658 . DOI:10.1021/ar4000339http://doi.org/10.1021/ar4000339 .

Li Yunkun(李芸焜), Li Yachao(李亚超), Xu Xianghui(徐翔晖), Gu Zhongwei(顾忠伟) . 高分子学报 , Acta Polymerica Sinica , 2017 . ( 2 ): 375 - 385.

Liu H, Wang H, Yang W, Cheng Y . J Am Chem Soc , 2012 . 134 17680 - 17687 . DOI:10.1021/ja307290jhttp://doi.org/10.1021/ja307290j .

Duan S, Yu B, Gao C, Yuan W, Ma J, Xu F J . ACS Appl Mater Interfaces , 2016 . 8 ( 43 ): 29334 - 29342 . DOI:10.1021/acsami.6b11029http://doi.org/10.1021/acsami.6b11029 .

Qi Y, Song H, Xiao H, Cheng G, Yu B, Xu F J . Small , 2018 . 14 ( 42 ): 1803061 DOI:10.1002/smll.v14.42http://doi.org/10.1002/smll.v14.42 .

Liao Z, Li Y, Lu H, Sung H . Biomaterials , 2014 . 35 ( 1 ): 500 - 508 . DOI:10.1016/j.biomaterials.2013.09.075http://doi.org/10.1016/j.biomaterials.2013.09.075 .

Tseng S, Liao Z, Kao S, Zeng Y, Huang K, Li H, Yang C, Deng Y, Huang C, Yang S, Yang P, Kempson I . Nat Commun , 2015 . 6 6456 DOI:10.1038/ncomms7456http://doi.org/10.1038/ncomms7456 .

Ping Y, Liu C D, Tang G P, Li J S, Li J, Yang W T, Xu F J . Adv Funct Mater , 2010 . 20 ( 18 ): 3106 - 3116 . DOI:10.1002/adfm.201000177http://doi.org/10.1002/adfm.201000177 .

更多指标>

浏览量

下载量

CSCD

文章被引用时，请邮件提醒。

提交

工具集

关联资源

含悬垂双键的可降解支化聚合物的合成与表征

正离子聚合制备具有双峰分布丁基橡胶及其结构性能研究

具生物活性的快速光交联生物可降解水凝胶的设计合成

双烯化合物类单体合成支化聚合物的支化结构的研究

单电子转移活性自由基聚合制备支化聚丙烯酸甲酯的研究