冬凌草甲素通过上调p53抑制三阴性乳腺癌细胞增殖并诱导凋亡的机制

Oronidin Inhibits Proliferation and Induces Apoptosis of Triple Negative Breast Cancer Cells by Upregulating p53

目的研究冬凌草甲素对三阴性乳腺癌细胞株MDA-MB-468增殖和凋亡的影响,并探索其潜在的作用机制。方法采用MTT法检测冬凌草甲素对乳腺癌细胞活力的影响,流式细胞分析细胞凋亡率,免疫印迹法检测细胞增殖及凋亡相关蛋白的表达。结果冬凌草甲素对三阴性乳腺癌细胞在各时间点存在不同程度的抑制作用,浓度10μM时,抑制率可达97%。随着冬凌草甲素作用浓度增加,光镜观察及DAPI染色均发现细胞数量显著减少。细胞周期结果提示,随着冬凌草甲素浓度提高,G2/M期细胞比例增加,早、晚期凋亡细胞比例增加。冬凌草甲素处理后,p53蛋白表达出现显著上调(P<0.05),同时PARP,Caspase-3以及Caspase-9的活化形式表达上调。结论冬凌草甲素能够有效抑制三阴性乳腺癌细胞MDA-MB-468增殖,阻滞细胞周期在G2/M期,促进细胞凋亡,并呈剂量依赖性,其作用机理与上调p53表达,活化PARP、Caspase-3和Caspase-9有关。

Objective To study the effect of oridonin on the proliferation and apoptosis of triple-negative breast cancer cell line MDA-MB-468 and explore the potential mechanism.Methods MTT assay was used to detect the viability of breast cancer cells after treatment with oridonin.Flow cytometry was used to analyze the apoptosis rate,and western blot was used to detect cell proliferation and apoptosis-related proteins.Results Oridonin inhibited the proliferation of triple-negative breast cancer cells at different time points.When the concentration reached 10μm,the inhibition rate was at 97%.Light microscopy and DAPI staining showed that the number of cells gradually decreased with the increment of oridonin.When the concentration of oridonin increased gradually,the proportion of the G2/M phase increased gradually,also the proportion of early and late apoptotic cells.After oridonin treatment,the expression of p53 protein was significantly up-regulated(P<0.05),and the expressions of PARP,caspase-3,and caspase-9 were up-regulated.Conclusion Oridonin can effectively inhibit MDA-MB-468 cells,arrest the cell cycle and induce apoptosis in a dose-dependent manner.The mechanism of action is related to the up-regulation of p53 expression and the activation of Caspase-3 and caspase-9.