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【論文摘要】Enhanced Proliferation of Endothelial Progenitor Cells Post Ultrasonic Microbubble Transfection is Plasmid DNA Size-dependent And Contributed by IL-6 Generation
摘要
Background/Synopsis: Ultrasonic microbubble transfection (UMT) has been shown to enhance in vitro and in vivo gene transfer. However, optimization of the method is necessary to facilitate the clinical application. There is convincing evidence that UMT enhances the efficiency of plasmid-mediated transfection in vitro and in vivo when using plasmids in the 3 - 5 kb size range. In addition, it has been recognized that the transfer of large DNA constructs into cells is important as maintaining the natural genomic environment of gene and providing tissue-specific regulation. Objectives/Purpose: We investigated whether Ultrasonic Microbubble Transfection (UMT) would enhance the transfection of large-sized luciferase plasmids (5.6, 9.2 and 33 kb) and biological impacts. Methods/Results: Endothelial progenitor cells (EPC) originated from porcine peripheral blood were cultured with medium containing constructed VEGF plasmid DNA (0.5 kb; CMV promoter) and air-gassed, hard shell microbubbles followed by UMT. Expression of VEGF was evaluated by ELISA 48 hours post gene delivery. In parallel, a series of parameters of ultrasound and functional evaluations were conducted to investigate the effects of UMT of VEGF gene into EPC. The results showed enhanced luciferase expression after UMT but the enhancement declined when the size of plasmid increased. UMT of pDNAs sized 5.6 and 9.2 kb into EPCs led to significant enhancement of proliferation. The secreted IL-6 from UMT of EPCs also increased in the 5.6 and 9.2 kb pDNA groups. Treatment of the transfected EPCs with anti-IL-6 antibody neutralized the proliferation. Conclusion: In conclusion, UMT of pDNAs sized 5.6 and 9.2 kb into EPCs increased the secretion of IL-6, which in turn enhanced cell proliferation.
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