冬凌草甲素固体脂质纳米粒干预食管癌细胞的增殖

Oridonin solid lipid nanoparticles inhibit proliferation of esophageal cancer cells

背景:越来越多的研究显示由中药制成的固体脂质纳米粒有抑制癌细胞增殖、诱导细胞凋亡的作用。目的:探讨冬凌草甲素固体脂质纳米粒对食管癌细胞增殖和凋亡的影响。方法:①0,2.5,5,10,20μmol/L冬凌草甲素固体脂质纳米粒作用于人食管癌Eca-109细胞24,48,72 h后,CCK8检测细胞抑制率,计算IC50;②0,14μmol/L冬凌草甲素固体脂质纳米粒作用人食管癌Eca-109细胞48 h后,流式细胞仪检测细胞凋亡率;③Western blot检测Cleaved caspase3、β-catenin、C-myc、Cyclin D1蛋白表达;④Wnt/β-catenin信号通路激活剂氯化锂及氯化锂+冬凌草甲素固体脂质纳米粒作用于人食管癌Eca-109细胞48 h后,再检测上述相关指标,对照组细胞不做任何处理。结果与结论:①不同浓度的冬凌草甲素固体脂质纳米粒作用于食管癌细胞24,48,72 h后的细胞抑制率均显著高于0 h时的细胞抑制率,且随着时间延长及浓度增加细胞抑制率增加(P<0.01);②14μmol/L组细胞凋亡率及Cleaved caspase3蛋白表达均显著高于0μmol/L组,β-catenin、C-myc、Cyclin D1蛋白表达均显低于0μmol/L组(P<0.01);③激活剂组及激活剂+冬凌草甲素固体脂质纳米粒组细胞抑制率、凋亡率及Cleaved caspase3蛋白表达均显著高于对照组,β-catenin、C-myc、Cyclin D1蛋白表达均显著低于对照组(P<0.01);④激活剂+冬凌草甲素固体脂质纳米粒组细胞抑制率、凋亡率及Cleaved caspase3蛋白表达显著低于激活剂组,β-catenin、C-myc、Cyclin D1蛋白表达显著高于激活剂组(P<0.01);⑤结果表明,冬凌草甲素固体脂质纳米粒可抑制人食管癌Eca-109细胞增殖并促进细胞凋亡,其机制与Wnt/β-catenin信号通路的调控有关。

BACKGROUND： Increasing studies have shown that solid lipid nanoparticles made from traditional Chinese medicinecan inhibit cancer cell proliferation and induce cell apoptosis.OBJECTIVE： To investigate the mechanisms of oridonin solid lipid nanoparticles （ORI-SLN） by the regulation ofWnt/β-catenin signaling pathway on the proliferation and apoptosis of esophageal cancer cells.METHODS： After 0, 2.5, 5, 10, 20 μmol/L ORI-SLN treated human esophageal cancer cell lines Eca-109 for 24, 48,72 hours, the cell inhibition rate was detected by cell counting kit-8, and the half maximal inhibitory concentration（IC50） was calculated. After 0, 14 μmol/L ORI-SLN treated Eca-109 cells for 48 hours, the cell apoptosis wasdetected by flow cytometry. The expression of Cleaved caspase3, β-catenin, C-myc, Cyclin D1 proteins wasdetected by western blot assay. Wnt/β-catenin signaling pathway activator LiCl and LiCl＋ORI-SLN were used totreat Eca-109 cells for 48 hours, and then the relevant indicators were reexamined. Eca-109 cells without any treatmentwere used as controls.RESULTS AND CONCLUSION： The cell inhibition rate of Eca-109 cells treated with different concentrations of ORI-SLNfor 24, 48 and 72 hours was significantly higher than that at 0 hour, and the cell inhibition rate was found to increase withthe prolongation of time and the increase of the concentration （P 〈 0.01）. 14 μmol/L ORI-SLN was confirmed to resultin the higher cell apoptosis and Cleaved caspase3 expression compared with the 0 μmol/L group, while theexpression of β-catenin, C-myc, Cyclin D1 proteins were significantly lower than the 0 μmol/L group （P 〈 0.01）. Cellinhibition rate, apoptosis rate and Cleaved caspase3 protein expression in the activator group andORI-SLN＋activator group were significantly higher than those in the control group, and the expression of β-catenin,C-myc, Cyclin D1 protein was significantly lower than those in the control group （P 〈 0.01）. The cell inhibition rate,apoptosis rate and expression of Cleaved caspase3 in ORI-SLN＋activator group was significantly lower than thosein the activator group, and the β-catenin, C-myc, Cyclin D1 protein expression was significantly higher than that inthe activator group （P 〈 0.01）. To conclude, ORI-SLNs can inhibit the proliferation and apoptosis of humanesophageal carcinoma cell line Eca-109, and its mechanism is related to the regulation of Wnt/β-catenin signalingpathway.