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功能性聚酰胺-胺树枝状大分子/凋亡素基因的多聚体在体外诱导人原发性胶质瘤细胞凋亡。

Polyplexes of Functional PAMAM Dendrimer/Apoptin Gene Induce Apoptosis of Human Primary Glioma Cells In Vitro.

作者信息

Bae Yoonhee, Thuy Le Thi, Lee Young Hwa, Ko Kyung Soo, Han Jin, Choi Joon Sig

机构信息

Department of Physiology, College of Medicine, Cardiovascular and Metabolic Disease Center, Inje University, Busan 47392, Korea.

Department of Biochemistry, College of Natural Sciences, Chungnam National University, Daejeon 305-764, Korea.

出版信息

Polymers (Basel). 2019 Feb 10;11(2):296. doi: 10.3390/polym11020296.

DOI:10.3390/polym11020296
PMID:30960280
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6419211/
Abstract

Highly efficient and safe gene delivery has become an important aspect of neuronal gene therapy. We evaluated the ability of polyamidoamine (PAMAM) dendrimer grafted with phenylalanine, histidine, and arginine (PAMAM-FHR), a nonviral gene delivery vector, to deliver a therapeutic, tumor cell-specific killer gene, apoptin, into the human primary glioma cell line GBL-14 and human dermal fibroblasts. We performed a transfection assay using plasmids of luciferase and enhanced green fluorescent protein (EGFP) and assessed cell viability. Both cell lines were treated with complexes of PAMAM-FHR and apoptin after which their intracellular uptake and localization were examined by fluorescence-activated cell sorting (FACS)analysis and confocal laser scanning microscopy. Confocal microscopy showed that the PAMAM-FHR escaped from the endo-lysosome into the cytosol. Cell cycle phase distribution analysis, annexin V staining, and a tetramethylrhodamine ethyl ester (TMRE) assay established that apoptin triggered apoptosis in the GBL-14 cell line but not in normal fibroblasts. These results indicated that the PAMAM-FHR/apoptin complex is an effective gene vehicle for cancer therapy in vitro.

摘要

高效且安全的基因传递已成为神经元基因治疗的一个重要方面。我们评估了接枝有苯丙氨酸、组氨酸和精氨酸的聚酰胺-胺（PAMAM）树枝状大分子（PAMAM-FHR），一种非病毒基因传递载体，将治疗性的、肿瘤细胞特异性杀伤基因凋亡素递送至人原发性胶质瘤细胞系GBL-14和人皮肤成纤维细胞的能力。我们使用荧光素酶和增强型绿色荧光蛋白（EGFP）质粒进行了转染试验，并评估了细胞活力。用PAMAM-FHR与凋亡素的复合物处理这两种细胞系，之后通过荧光激活细胞分选（FACS）分析和共聚焦激光扫描显微镜检查它们的细胞内摄取和定位。共聚焦显微镜显示PAMAM-FHR从内溶酶体逃逸到细胞质中。细胞周期阶段分布分析、膜联蛋白V染色和四甲基罗丹明乙酯（TMRE）检测确定凋亡素在GBL-14细胞系中触发了凋亡，但在正常成纤维细胞中未触发。这些结果表明PAMAM-FHR/凋亡素复合物是一种用于体外癌症治疗的有效基因载体。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e441/6419211/6aef90357966/polymers-11-00296-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e441/6419211/5ddc5e6e7994/polymers-11-00296-sch001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e441/6419211/3b0a3a9218ae/polymers-11-00296-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e441/6419211/ed09ae5f858e/polymers-11-00296-g002a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e441/6419211/5cf1824a50a4/polymers-11-00296-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e441/6419211/4d13a4a12bcd/polymers-11-00296-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e441/6419211/1305974806ff/polymers-11-00296-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e441/6419211/8ad80f4c447c/polymers-11-00296-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e441/6419211/5a194ff332db/polymers-11-00296-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e441/6419211/096075297b4d/polymers-11-00296-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e441/6419211/6aef90357966/polymers-11-00296-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e441/6419211/5ddc5e6e7994/polymers-11-00296-sch001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e441/6419211/3b0a3a9218ae/polymers-11-00296-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e441/6419211/ed09ae5f858e/polymers-11-00296-g002a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e441/6419211/5cf1824a50a4/polymers-11-00296-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e441/6419211/4d13a4a12bcd/polymers-11-00296-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e441/6419211/1305974806ff/polymers-11-00296-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e441/6419211/8ad80f4c447c/polymers-11-00296-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e441/6419211/5a194ff332db/polymers-11-00296-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e441/6419211/096075297b4d/polymers-11-00296-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e441/6419211/6aef90357966/polymers-11-00296-g009.jpg

相似文献

[1]
Polyplexes of Functional PAMAM Dendrimer/Apoptin Gene Induce Apoptosis of Human Primary Glioma Cells In Vitro.

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[3]
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[4]
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[5]
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[6]
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[5]
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[6]
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[7]
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[8]
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[9]
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[10]
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本文引用的文献

[1]
Efficient Binding, Protection, and Self-Release of dsRNA in Soil by Linear and Star Cationic Polymers.

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Polymers (Basel). 2018-4-1

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Apoptin Gene Delivery by the Functionalized Polyamidoamine Dendrimer Derivatives Induces Cell Death of U87-MG Glioblastoma Cells.

J Pharm Sci. 2017-6

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Dipeptide-functionalized polyamidoamine dendrimer-mediated apoptin gene delivery facilitates apoptosis of human primary glioma cells.

Int J Pharm. 2016-12-30

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Int J Pharm. 2015-8-15

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