丹酚酸B及其磷脂复合纳米粒抑制口腔鳞癌和白斑细胞株的作用

Salvianolic Acid B Phospholipid Complex Loaded Nano Particles Induces Apoptosis in OSCC HN13 and Oral Leukoplakia Leuk1 Cells

目的:研究丹酚酸B(Sal B)及其磷脂复合纳米粒(Sal B-PLC-NPs)对口腔鳞癌HN13和白斑Leuk1的抑制活性。方法:利用激光共聚焦显微镜、荧光分光光度计、MTS法、流式细胞仪分析Sal B-PLC-NPs与Sal B影响HN13、Leuk1摄取药物和细胞活性的差异。结果:200μg/mL Sal B-PLC-NPs作用2 h,显示胞内更多、更强的蓝绿色荧光,提示Sal B-PLC-NPs促进细胞摄取药物。Sal B-PLC-NPs对25、50、100、200μg/mL HN13及25、200μg/mL Leuk1作用2 h,胞内荧光更强,差异有统计学意义(P<0.05)。相比空白组,Sal B-PLC-NPs和Sal B均抑制细胞增殖;Sal B-PLC-NPs对HN13(200μg/mL,48 h)抑制增殖更强,对Leuk1(200μg/mL,96 h)抑制作用强于Sal B。Sal B-PLC-NPs对HN13(200μg/mL,24 h)周期阻滞更强,而Sal B对Leuk1(200μg/mL,48 h)阻滞比Sal B-PLC-NPs明显。Sal B诱导HN13(200μg/mL,24 h)及Leuk1(200μg/mL,48 h)凋亡率比Sal B-PLC-NPs更高。阻滞周期、诱导凋亡结果与细胞抑制曲线相符。结论:本实验证实,Sal B-PLC-NPs被细胞摄取量更大,进而提高了Sal B对HN13、Leuk1抑制增殖、阻滞周期和诱导凋亡的活性,呈时间-浓度依赖性,Sal B-PLC-NPs有望成为防治白斑癌变的新剂型。

Object:The purpose of the study was to compare anticancer effects between Salvianolic acid B phospholipid complex loaded nano particles(Sal B-PLC-NPs)and Salvianolic acid B(Sal B)in OSCC cell line HN13 and Oral Leukoplakia Leuk1 cells.Methods:Cells were treated with 200μg/m of Sal B-PLC-NPs and Sal B for 2 hours and photographed by laser confocal microscopy and quantified.Cells were placed in plates and Sal B-PLC-NPs and Sal B were added at final concentrations of 25,50,100,200μg/mL and incubated for 2 hours.At 200μg/mL concentration,incubation time was set at 5,10,20.30,60,120 min respectively.Cell viability was determined by flow cytometric procedures.MTS assay was used to analyze differences in inhibition of cell proliferation,retardation cell cycle and induction cell apoptosis.Results:Cells treated with Sal B-PLC-NPs intuitively showed stronger fluorescence than cells treated with Sal B,which proved Sal B-PLC-NPs promoted drug to be absorbed by cells.Sal B-PLC-NPs was more absorbable by HN13 at 25,50,100 and 200μg/mL concentrations.Compared with blank control group,Sal B-PLC-NPs and Sal B inhibition cell proliferation all had statistical difference.Sal B-PLC-NPs inhibition cell proliferation about HN13(200μg/mL,48 h)and Leuk1(200μg/mL,96 h)was more significant than Sal B.Sal B-PLC-NPs arrest cell cycle of HN13(200μg/mL,24 h)showed more obvious trend than Sal B.However,Sal B arrest cell cycle of Leuk1(200μg/mL,48 h)was more significant than Sal B-PLC-NPs.Sal B inducing apoptosis about HN13(200μg/mL,24 h)and Leuk1(200μg/mL,48 h)were more obvious than Sal B-PLC-NPs.The last two results were consistent with curves of cell growth inhibition.Conclusion:The assay confirmed that Sal B-PLC-NPs showed more absorption by cells and better bioactivity about inhibition of proliferation,arrest cycle and induction apoptosis of HN13 and Leuk1 in a time-and dose-dependent manner than Sal B.