[关键词]
[摘要]
目的 研究黄芩素对鼻咽癌CNE2细胞增殖和凋亡的影响,并探讨其阻滞细胞周期和诱导凋亡的机制。方法 通过CCK-8法和克隆形成实验检测黄芩素对CNE2细胞增殖的影响,流式细胞仪检测细胞凋亡率和细胞周期变化,荧光显微镜观察细胞形态改变,试剂盒检测Caspase-3蛋白活性变化,实时荧光定量PCR(qRT-PCR)检测P53、P21、CDK2、Caspase-3 mRNA表达情况。结果 黄芩素对鼻咽癌CNE2细胞增殖具有显著的抑制作用,且呈剂量和时间依赖性,与对照组比较,不同浓度的黄芩素作用于CNE2细胞48 h后,细胞出现典型的凋亡特征,并出现周期阻滞,Caspase-3活性增强,P53及其下游基因P21、Caspase-3 mRNA表达水平增高,CDK2 mRNA表达水平下降。结论 黄芩素可抑制鼻咽癌CNE2细胞增殖,可能是通过上调P53表达诱导细胞凋亡并阻滞细胞周期于S期。
[Key word]
[Abstract]
Objective To investigate the anti-proliferation effect of baicalein in nasopharyngeal carcinoma CNE2 cells and to clarify its mechanisms of cell cycle arrest and apoptosis. Methods The proliferative activity was detected by CCK-8 assay, and the changes of cell morphology was observed under fluorescence microscope. The rate of apoptosis and the cycle arrest were detected by flow cytometry, and caspase-3 activity was detected by caspase-3 activity kit. Expression of related genes (P53, P21, CDK2, and Caspase-3) at the RNA level were detected by using qRT-PCR. Results Baicalein had a significant inhibitory proliferation effect on CNE2 cells, which was the dose and time-dependent. After 48 h treatment with different concentrations of baicalein, the cells showed obvious apoptotic characteristics, with the increase of apoptotic cells, the activity of caspase-3 was increased, and S phase cells increase. Compared with the control group, the mRNA expression of P53, P21 and Caspase-3 increased, and mRNA expression of CDK2 decreased. Conclusion Baicalein could inhibit the proliferation of nasopharyngeal carcinoma CNE2 cells, and may induce apoptosis and cell cycle arrest in S phase by up regulating the expression of P53.
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[基金项目]
国家自然科学基金资助项目(81260405);广西自然科学基金项目(2015GXNSFAA139215);广西生物靶向诊治研究重点实验室开放课题(GXSWBX201502)