摘要
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。
Objective：To analyze the expression of mitochnodria pathway-related gene and ex- plore 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 concen- trations （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 easpase-3, caspase-9 mRNA were increased by POD- NLC too. Meanwhile the expression of bcl-2 mRNA decreased. The difference wes statistically sig- nificant 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-NLCmight induce the apoptosis of VK2/E6E7 cells through mitochnodria pathway.
出处
《皮肤性病诊疗学杂志》
2013年第2期85-88,共4页
Journal of Diagnosis and Therapy on Dermato-venereology
基金
国家自然科学基金项目(编号:81171627)
广东省教育厅科技创新重点项目(编号:cxzd1119)
