The role of caveolin-1 and syndecan-4 in the internalization of PEGylated PAMAM dendrimer polyplexes into myoblast and hepatic cells.
The role of caveolin-1 and syndecan-4 in the internalization of PEGylated PAMAM dendrimer polyplexes into myoblast and hepatic cells.
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DOI：
10.1016/j.ejpb.2014.07.010
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发表时间：
2014-11
期刊：
影响因子：
--
通讯作者：
Qi R
中科院分区：
文献类型：
--
作者：
Shen W;van Dongen MA;Han Y;Yu M;Li Y;Liu G;Banaszak Holl MM;Qi R
为了提高聚乙二醇化聚酰胺胺树突状大分子在肝脏和肌肉中的基因传递效率，在C2C12和HepG2细胞中探讨了Syndecan-4受体和小窝蛋白-1蛋白在聚乙二醇化第五代(G5-PEG)和第七代(G7-PEG)树突状大分子和质粒DNA复合体内吞作用中的作用。通过将Syndecan-4特异性siRNA导入细胞，两种细胞的Syndecan-4表达水平均下调。用腺病毒载体表达小窝蛋白-1(Ad-CAV-1)感染细胞，上调小窝蛋白-1的表达。用流式细胞仪检测Syndecan-4和Caveolin-1对G5-PEG/DNA或G7-PEG/DNA复合体内吞作用的影响。我们的结果表明，在HepG2细胞中，Syndecan-4的下调和Caveolin-1的上调显著地促进了PEG-PAMAM树枝状大分子多聚体的内化；而在C2C12细胞中，Syndecan-4的下调减少了多聚体的内化，而Caveolin-1的上调对内化没有影响。G5-PEG/pGFP在两种细胞系上的基因表达结果显示，Syndecan-4和Caveolin-1的基因表达趋势与观察到的复合体内吞作用的趋势相同。本研究为临床转基因治疗中如何通过上调或下调Syndecan-4和Caveolin-1的表达来提高聚乙二醇-PAMAM树状大分子的体内基因输送效率提供了线索。
To improve gene delivery efficiency of PEGylated poly(amidoamine) dendrimers in livers and muscles, the roles of syndecan-4 receptor and caveolin-1 protein in the endocytosis of PEGylated generation 5 (G5-PEG) or 7 (G7-PEG) dendrimers and plasmid DNA polyplexes were explored in C2C12 and HepG2 cells. Expression levels of syndecan-4 for both cell lines were downregulated by transfection of the cells with syndecan-4 specific siRNA. Caveolin-1 was upregulated by infecting the cells with adenovirus vector expressed caveolin-1 (Ad-CAV-1). The impact of syndecan-4 and caveolin-1 on endocytosis of G5-PEG/DNA or G7-PEG/DNA polyplexes was then measured by flow cytometry. Our results demonstrate that downregulation of syndecan-4 and upregulation of caveolin-1 significantly improved internalization of PEG-PAMAM dendrimer polyplexes in HepG2 cells; however, in C2C12 cells, downregulation of syndecan-4 decreased the internalization of the polyplexes while upregulation of caveolin-1 had no effect on internalization. Gene expression results for G5-PEG/pGFP on the two cell lines exhibited the same trends for syndecan-4 and caveolin-1 as was observed for endocytosis of the polyplexes. This study gives a clue how to take strategies by up- or down-regulation of the expressions of Syndecan-4 and Caveolin-1 to improve in vivo gene delivery efficiency of the PEG-PAMAM dendrimers in clinical transgenic therapy.
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DOI：
10.1111/febs.12072
发表时间：
2013-02
期刊：
The FEBS journal
影响因子：
--
作者：
Jackson MF;Hoversten KE;Powers JM;Trobridge GD;Rodgers BD
通讯作者：
Rodgers BD
DOI：
10.1073/pnas.85.24.9562
发表时间：
1988-12-01
影响因子：
11.1
作者：
SANDERSON, RD;BERNFIELD, M
通讯作者：
BERNFIELD, M
影响因子：
3.5
作者：
Kopatz, I;Remy, JS;Behr, JP
通讯作者：
Behr, JP
影响因子：
4.9
作者：
Rattan R;Vaidyanathan S;Wu GS;Shakya A;Orr BG;Banaszak Holl MM
通讯作者：
Banaszak Holl MM
影响因子：
4.8
作者：
Paris, Sebastien;Burlacu, Alina;Durocher, Yves
通讯作者：
Durocher, Yves