Rac-Mediated Macropinocytosis Is a Critical Route for Naked Plasmid DNA Transfer in Mice
Rac-Mediated Macropinocytosis Is a Critical Route for Naked Plasmid DNA Transfer in Mice
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DOI：
10.1021/mp900042p
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发表时间：
2009-07-01
影响因子：
4.9
通讯作者：
Nishida, Koyo
中科院分区：
文献类型：
--
作者：
Fumoto, Shintaro;Nishi, Junya;Nishida, Koyo
我们最近发现了体内裸质粒DNA（pDNA）转移到小鼠胃浆膜表面细胞中的潜力。由于pDNA是巨大的分子，无载体和物理力的基因转移机制具有极大的生物学意义。裸pDNA进入胃间皮细胞的内吞途径不是网格蛋白或小窝介导的内吞，而是巨胞饮。裸pDNA转移需要肌动蛋白聚合和基于肌球蛋白的收缩。上游激酶Rho家族GTP酶、Syk、Src家族激酶和Pl-3 K参与裸pDNA转移。此外，细胞内信号通路不是通过Rho通路介导的，而是通过Rac通路介导的。Rac、PAK和WAVE 2的下游分子与裸pDNA转移协同作用。总之，证明了Rac信号传导途径调节裸pDNA的巨胞饮作用。本研究的结果将有助于阐明体内细胞功能和提高体内转染效率。
We have recently discovered the potential for in vivo naked plasmid DNA (pDNA) transfer into gastric serosal surface cells in mice. As pDNA are huge molecules, the mechanism of gene transfer without carriers and physical forces is of great biological interest. The endocytic route for naked pDNA transfer into gastric mesothelial cells was not clathrin- or caveolaemediated endocytosis, but macropinocytosis. Naked pDNA transfer required both actin polymerization and myosin-based contraction. Upstream kinases of Rho family GTPases, Syk, Src family kinases and Pl-3K were involved in naked pDNA transfer. Furthermore, the intracellular signaling pathway was not mediated via the Rho pathway, but by the Rac pathway. Downstream molecules of Rac, PAK and WAVE2 co-operated with naked pDNA transfer. Overall, it was demonstrated that the Rac signaling pathway regulated the macropinocytosis of naked pDNA. The information in this study would be helpful to clarify in vivo cell functions and to improve in vivo transfection efficiency.