鬼臼毒素纳米脂质载体通过非Caspase依赖途径诱导VK2/E6E7细胞凋亡机制的研究

The Research of Apoptosis Mechanism Induced by Podophyllotoxin Nanostructured Lipid Carriers in VK2/E6E7 Cells through Caspaseindependent Pathway

目的:探讨鬼臼毒素纳米脂质载体(POD-NLC)诱导人永生化阴道上皮细胞(VK2/E6E7)凋亡的作用及机制。方法:取对数期生长VK2/E6E7细胞,分别加入POD-NLC和Caspase抑制剂Z-VAD-FMK共同培养24小时。实验分为4组:A组:空白对照组;B组:0.25μg/mL POD-NLC+Caspase抑制剂组;C组:1μg/mL POD-NLC+Caspase抑制剂组;D组:空白NLC+Caspase抑制剂组。采用流式细胞仪检测ROS水平,qPCR检测AIF mRNA表达,蛋白印迹检测各组细胞AIF、cyt-C蛋白表达。结果:B组、C组均可以上调ROS、AIF、cyt-C水平,与A组及D组相比均有统计学差异(P值均<0.05);B组与C组AIF mRNA及AIF蛋白表达无统计学差异(P值均>0.05);ROS、cyt-C各组间比较均有统计学意义(P值均<0.05)。结论:鬼臼毒素纳米脂质载体可以通过非Caspase依赖途径诱导永生化阴道上皮细胞凋亡。

Objective：To investigate the apoptosis-inducing effect and its mechanism of Podo- phyllotoxin Nanostrnetured Lipid Carriers （ POD-NLC ） in VK2/E6E7 cells. Methods： The VK2/ E6E7 cells were divided into 4 groups ： group A ： control ; group B ： 0. 25μg/mL POD-NLC with Caspase inhibitor; group C： 1 μg/mL POD-NLC with Caspase inhibitor; group D： control NLC with Caspase inhibitor. The ROS were measured by flow cytometry, expression of AIF mRNA was detected by qPCR, and expression of AIF and cyt-C was examined by Western blot. Results：The group B and C showed the increasing of expression of ROS, AIF and eyt-C, and showed the statis- tical significant compared with group A and D （P 〈 0. 05 ）. The expression of AIF mRNA, AIF in group B and group C has no statistical difference （P 〉 0. 05 ）. There were statistical difference on the ROS and cyt-C （ P 〈 0. 05 ） in three study groups. Conclusion： Podophyllotoxin nanostrnctured lipid carriers might induce VK2/E6E7 cells apoptosis through caspase-independent pathway.