CAJ | 학술논문

目的观察自制阳离子纳米泡作为基因转染载体体外转染肝癌细胞系HepG2细胞的可行性.方法将脂质体、聚乙烯亚胺(PEI)和全氟丙烷声振后制得微囊带有PEI的阳离子脂质体纳米泡(PNB),评价其基本特性,用其转染HepG2细胞,并辐照超声,荧光显微镜观察绿色荧光蛋白表达情况,流式细胞仪检测细胞转染率,CCK-8法评估其细胞活性.结果制备的PNB表面带有正电荷,能阻碍质粒DNA在电场作用下发生迁移,且能有效地转染HepG2细胞,转染效率达30％～45％,辐照超声后转染效率明显提高(P＜0.05).结论新型阳离子纳米泡在体外可作为基因转染载体,有效地携带质粒DNA并促进转染,辐照超声可使转染效率显著提高.
목적 관찰자제양리자납미포작위기인전염재체체외전염간암세포계HepG2세포적가행성.방법 장지질체、취을희아알(PEI)화전불병완성진후제득미낭대유PEI적양리자지질체납미포(PNB),평개기기본특성,용기전염HepG2세포,병복조초성,형광현미경관찰록색형광단백표체정황,류식세포의검측세포전염솔,CCK-8법평고기세포활성.결과 제비적PNB표면대유정전하,능조애질립DNA재전장작용하발생천이,차능유효지전염HepG2세포,전염효솔체30％～45％,복조초성후전염효솔명현제고(P＜0.05).결론 신형양리자납미포재체외가작위기인전염재체,유효지휴대질립DNA병촉진전염,복조초성가사전염효솔현저제고.
Objective To observe self-made cationic nanobubbles as non-viral gene carrier to transfer green fluorescent protein reporter gene into HepG2 cell in vitro.Methods Cationic nanobubbles(PNB) were prepared by sonicating liposomes、polyethylenimine and perfluoropropan.The surface potential and the size of nanobubbles were assessed by laser particle analyzer.HepG2 cells were incubated with DNA,nanobubbles with or without ultrasound exposure.The transfection efficiency was evaluated by flow cytometer and the cell viability by cell counting Kit-8.Results The mean diameter of PNB was (834.57 ± 6.4) nm and the surface charge was (4.15± 0.98)mV.The PNB-DNA complexes,which blocked by the Agarose gel electrophoresis,could effectively transfer HepG2 cells,and the ultrasound exposure could enhance the transfection efficiency further significantly (P ＜ 0.05).Conclusions The new PNB could effectively combine with pDNA to enhance gene delivery and ultrasound exposure could improve its efficiency further in HepG2 cell in vitro.