隐丹参酮诱导乳腺癌MDA-MB-231细胞凋亡的研究

Study of cryptotanshinone inducing apoptosis of breast cancer MDA-MB-231 cells

目的探讨隐丹参酮对乳腺癌MDA-MB-231细胞凋亡的影响及其机制。方法用不同浓度的隐丹参酮处理MDA-MB-231细胞24 h。采用MTT、流式细胞术检测不同浓度隐丹参酮对MDA-MB-231细胞活性、凋亡的影响。采用Western blot检测MDA-MB-231细胞中Bcl-2、Bax、Caspase-3蛋白表达水平。结果与0μmol/L对照组相比,10μmol/L、20μmol/L、40μmol/L、80μmol/L隐丹参酮组MDA-MB-231细胞存活率显著降低,且呈剂量依赖(P<0.05)。流式细胞仪检测显示隐丹参酮在20μmol/L浓度时MDA-MB-231细胞凋亡率为(6.34±0.52)%,与对照组(6.09±0.76)%相比无统计学差异(P>0.05)。在40μmol/L、80μmol/L浓度时MDA-MB-231细胞凋亡率分别是(18.74±0.65)%、(28.04±3.08)%,与对照组相比有统计学差异(P<0.05),且两浓度组之间相比差异亦有统计学意义(P<0.05)。Western blot结果显示,与对照组相比,隐丹参酮在40μmol/L、80μmol/L浓度时,MDA-MB-231细胞内抗凋亡蛋白Bcl-2表达显著下调(P<0.05),同时,促凋亡蛋白Bax、Caspase-3表达显著增加(P<0.05)。结论隐丹参酮可诱导三阴乳腺癌MDA-MB-231细胞凋亡,其机制可能与激活Bax、Caspase-3和抑制Bcl-2等凋亡调控基因有关。

Objective To investigate the effect of cryptotanshinone on the apoptosis of breast cancer MDA-MB-231 cells and its mechanism. Methods MDA-MB-231 cells were treated with different concentrations of cryptotanshinone for 24h. The effects of different concentrations of cryptotanshinone on the activity and apoptosis of MDA-MB-231 cells were detected by MTY and flow cytometry. Western blot was used to detect the protein expressions of Bcl-2, Bax and Caspase-3 in MDA-MB-231 cells. Results Compared with that of the 0μmol/L control group, the survival rates of MDA-MB-231 cells in 10, 20, 40, 80 μmol/L cryptotanshinone groups were significantly decreased （P〈0.05）. And the survival rate was dose-dependent.Flow cytometry showed that the apoptosis rate of MDA-MB-231 cells was（6.34＋ 0.52）% at cryptotanshinone concentration of 20 μmol/L, which was not significantly different from that of the control group （6.09±0.76）% （P〉0.05）. The apoptosis rates of MDA-MB-231 cells were （18.74±0.65）% and （28.04±3.08）% respectively at 40 μmol/L and 80μmol/L, which were different from those of the control group （P〈0.05）, and the difference between the two concentration groups was statistically significant（P〈0.05）. Western blot results showed that antiapoptotic protein Bcl-2 expression in MDA-MB-231 ceils was significantly down-regulated at cryptotanshinone con centrations of 40 μmol/L and 80 μmol/L, compared with that of the control group（P〈0.05）. Meanwhile, the expression of pro-apoptotic proteins Bax and Caspase-3 increased significantly（P〈0.05）. Conclusion Cryptotanshinone can induce the apoptosis of triple negative MDA-MB-231 cells, which may be related to the activation of apoptosis regulatory genes including Bax,Caspase-3 and Bcl-2.