鬼臼毒素纳米脂质载体通过线粒体途径诱导人阴道上皮细胞凋亡的机制研究

目的:研究鬼臼毒素纳米脂质载体(POD-NLC)对体外培养的永生化人阴道上皮细胞(VK2/E6E7细胞)线粒体途径相关凋亡基因表达的影响,探索POD-NLC诱导VK2/E6E7细胞凋亡的作用机制。方法:自制0.5%POD-NLC,用不同浓度的POD-NLC(0.25,1μg/mL)和空白NLC作用VK2/E6E7细胞24 h后,采用实时荧光定量核酸扩增法(Real-time Quantitative PCR,qPCR)检测bcl-2、bax、caspace-3、caspase-9等mRNA表达水平,Western Bolt检测bcl-2、bax蛋白的表达水平。结果:POD-NLC呈剂量依赖性的上调bax mRNA的表达水平(P值均<0.01),同时也增加caspase-3、caspase-9 mRNA的表达(P值均<0.05),并降低bcl-2 mRNA表达量(P值均<0.01);空白-NLC组对上述mRNA的影响均无统计学意义(P值均>0.05)。POD-NLC可剂量依赖性地上调bax蛋白表达量(P值均<0.01),同时下调bcl-2蛋白表达量(P值均<0.01),但各药物处理组之间差异无统计学意义(P值均>0.05)。结论:POD-NLC能通过线粒体途径诱导VK2/E6E7细胞发生凋亡。

The Investigation on VK2/E6E7 Cells Apoptosis Induced by Podophyllotoxin Nanostructured Lipid Carriers through Mitochnodria Pathway

Objective:To analyze the expression of mitochnodria pathway-related gene and explore the possible apoptosis mechanisms by podophyllotoxin nanostructured lipid carriers(POD-NLC)on VK2/E6E7 cells in vitro.Methods:VK2/E6E7 cells were treated with different concentrations(0.25,1 μg/mL) of POD-NLC and blank-NLC.Bcl-2,bax,and caspace-3,caspase-9 mRNA were assessed by Real-time Quantitative PCR(qPCR),and Western Blot was employed to determine bcl-2,bax protein.Results:The results of qPCR showed that the levels of bax mRNA increased in a dose-dependent manner and caspase-3,caspase-9 mRNA were increased by POD-NLC too.Meanwhile the expression of bcl-2 mRNA decreased.The difference wes statistically significant compared with control group.The expression of bax protein significantly increased with the the dosage of POD-NLC.The expression of bcl-2 protein was significantly decreased by POD-NLC.However,there was no statistical difference between study groups.Conclusion:POD-NLC might induce the apoptosis of VK2/E6E7 cells through mitochnodria pathway.