【摘要】 目的探讨阳离子脂质体Lipofectamine2000(Lipo)在一定浓度下对细胞毒性的作用机制。方法应用一定毒性浓度的Lipo作用于胰腺癌Capan-2细胞,利用细胞直接记数法和流式细胞术检测其对Capan-2细胞生长、细胞凋亡和周期的影响。结果Lipo与siRNA的浓度均影响转染率的高低。在2ml转染体积中,Lipo在5μl的浓度下,使Capan-2细胞生长明显减慢,以第3天后更加明显(P<0.001)。随着转染细胞按常规培养的时间延长,早期凋亡细胞明显增多(P<0.05),活性细胞明显减少(P<0.01),而损伤细胞和死亡细胞明显增多(P<0.001),以48h后更明显;G0～G1期细胞明显增多(P<0.05),G2-M期细胞明显减少(P<0.01),S期细胞明显减少(P<0.01),晚期凋亡细胞减少(P<0.05)。结论在一定浓度下,Lipo可影响细胞的生长、早期凋亡和细胞周期分布,因此,利用Lipo在不同的细胞系进行基因转染过程中Lipo应进行浓度梯度筛选。更多还原

【Abstract】 Objective To investigate the toxicity of cationic liposome Lipofectamine 2000 (Lipo) in human pancreatic cancer Capan-2 cells.Methods Capan-2 cells were cultured in the presence of Lipo at toxic concentrations,and the cell growth,apoptosis and cell cycle changes were evaluated by cell counting and flow cytometry.Results The concentrations of both Lipo and siRNA affected the transfection efficiency.In a transfection volume of 2 ml,the presence of 5 μl Lipo resulted in slowed growth of Capan-2 cells,which was especially obvious after 3 days (P<0.001).Prolonged culture of the transfected cells caused significant increases in early apoptotic cells (P<0.05) and in the damaged or necrotic cells (P<0.001),and resulted in reduced viable cells (P<0.01); these changes became obvious after a 48-hour culture,which also increased the ratio of G0/G1 phase cells (P<0.05) and decreased those of G2/M phase cells (P<0.01),S phase cells (P<0.01),and the late apoptotic cells (P<0.05).Conclusion Toxic concentrations of Lipo can affect the growth,apoptosis and cell cycles of Capan-2 cells in vitro,and this urges careful concentration selection when using Lipo for gene transfer into different cells.更多还原