茶黄素对人鼻咽癌CNE2细胞增殖及凋亡的影响

目的:探究茶黄素对人鼻咽癌CNE2细胞增殖和凋亡的影响及其可能机制。方法:采用不同浓度茶黄素处理人鼻咽癌CNE2细胞;使用CCK-8法检测CNE2细胞在24、48、72 h的增殖抑制情况并计算细胞抑制率;细胞平板克隆形成实验观察CNE2细胞的克隆形成能力;细胞划痕实验检测CNE2细胞的迁移情况;Hoechst 33258染色和流式细胞术检测CNE2细胞凋亡情况;实时荧光定量PCR检测相关基因的表达水平。结果:CCK-8检测结果显示随着茶黄素药物浓度的升高及作用时间的延长,药物对CNE2细胞的增殖有明显的抑制作用,其作用呈时间及浓度依赖性;细胞平板克隆实验显示茶黄素能显著降低CNE2细胞的克隆形成能力;Hoechst33258染色显示茶黄素作用于CNE2细胞后出现核固缩、碎裂等凋亡现象;细胞流式术检测显示经茶黄素处理后CNE2细胞凋亡率升高;RT-PCR显示茶黄素能上调CNE2细胞Bax、Bcl-2、Caspase-3、P21 mRNA的表达。结论:茶黄素能抑制人鼻咽癌CNE2细胞增殖,并诱导其凋亡,其作用呈时间及浓度依赖性,其作用机制可能与调节细胞增殖和凋亡基因的表达有关。

Effect of Theaflavin on Proliferation and Apoptosis of Human Nasopharyngeal Carcinoma CNE2 Cells

Objective:To investigate the effects of theaflavin on proliferation and apoptosis of human nasopharyngeal carcinoma CNE2 cells and its possible mechanism.Methods:Human nasopharyngeal carcinoma CNE2 cells were treated with different concentrations of theaflavin.The proliferation inhibition of CNE2 cells at 24,48,72 h were inspected and the cell inhibition rate was calculated by CCK-8 assay.The clonal formation ability of CNE2 cells was observed by cell plate clonal formation experiments.The migration of CNE2 cells was detected by cell scratch test.The apoptosis of CNE2 cells was detected by Hoechst 33258 staining and flow cytometry.The gene expression levels of the related genes were detected by using real-time fluorescent quantitative PCR.Results:The results of CCK-8 showed that with the increase of the concentration of theaflavin and the prolongation of the action time,the proliferation of CNE2 cells was significantly inhibited,which was the time-concentration dependence.Cell plate clonal experiments showed that theaflavin significantly reduced the colony forming ability of CNE2 cells.The Hoechst 33258 staining showed the apoptosis such as nuclear pyknosis and fragmentation occurred after the treatment of CNE2 cells with theaflavin,and flow cytometry showed that the apoptosis rate of CNE2 cells increased after theaflavin treatment.RT-PCR showed that theaflavin up-regulated the mRNA expressions of the Bax,Bcl-2,Caspase-3,P21 in CNE2 cells.Conclusion:Theaflavin can inhibit the proliferation of human nasopharyngeal carcinoma CNE2 cells,and induce apoptosis with the time-concentration dependence,which may be related to the regulation of cell proliferation and expression of apoptotic genes.