柯里拉京体内外诱导口腔鳞癌CAL-27细胞凋亡及机制探讨

Corilagin-induced apoptosis of oral squamous carcinoma CAL-27 cells in vitro and in vivo and its mechanism

目的:观察柯里拉京(corilagin)对人舌鳞癌CAL-27细胞增殖和凋亡的作用,探讨诱导其凋亡的分子机制。方法:体外实验采用不同浓度的柯里拉京预处理CAL-27细胞,通过CCK-8法及平板克隆形成实验检测细胞增殖能力;流式细胞术检测细胞凋亡;qRT-PCR和Western印迹法检测对细胞中Bax、Bcl-2、Caspase-3、Cleaved Caspase-3mRNA及蛋白表达水平的影响。通过CAL-27细胞构建裸鼠移植瘤模型,检测柯里拉京的体内抗肿瘤效果。采用GraphPad Prism 8.0软件包对数据进行统计学分析。结果:体外实验结果显示,柯里拉京以浓度依赖性方式抑制CAL-27细胞增殖,诱导细胞凋亡;在mRNA和蛋白水平上上调Bax、Caspase-3和Cleaved Caspase-3,下调Bcl-2,差异均有统计学意义(P<0.05)。体内实验表明,与对照组相比,柯里拉京组可显著减小裸鼠瘤体体积(P<0.05)。结论:柯里拉京在体内和体外均能显著抑制人舌鳞癌CAL-27细胞增殖并促进其凋亡,其作用机制可能与调控Bax/Bcl-2/Caspase-3信号通路有关。

PURPOSE:To investigate the effect of corilagin on proliferation and apoptosis of human oral squamous cell carcinoma CAL-27 cells,and to explore the molecular mechanism of inducing cell apoptosis.METHODS:In vitro experiments,Cal-27 cells were treated with different concentrations of corilagin,cell-counting kit-8(CCK-8)assay and colony formation assay were performed to evaluate cell proliferation;flow cytometric analysis was used to evaluate cell apoptosis;qRT-PCR and Western blot assays were performed to evaluate the effect of corilagin on the expression levels of Bax,Bcl-2,Caspase-3,Cleaved Caspase-3 in CAL-27 cells.In vivo experiments,tumor-bearing nude mice was constructed with CAL-27 cells to evaluate the antitumor effect of corilagin.GraphPad Prism 8.0 software package was used for statistical analysis of the data.RESULTS:In vitro experiments showed that corilagin in a dose-dependent manner inhibited proliferation,induced apoptosis,up-regulated Bax,caspase-3,cleaved caspase-3 and down-regulated Bcl-2 at the mRNA and protein levels of CAL-27 cells,and the differences were statistically significant(P<0.05).In vivo experiments showed that compared with the control group,corilagin could significantly reduce the volume of tumor in nude mice(P<0.05).CONCLUSIONS:Corilagin can significantly inhibit CAL-27 cell growth and promote its apoptosis both in vitro and in vivo,which may be related to the mediation of Bax/Bcl-2/Caspase-3 signaling pathway.