迷迭香酸对乳腺癌MDA-MB-231细胞增殖、凋亡和迁移能力的影响

Effects of Rosmarinic Acid on Cell Proliferation,Apoptosis and Migration in Breast Cancer MDA-MB-231 Cells

研究迷迭香酸对乳腺癌MDA-MB-231细胞增殖、凋亡和迁移能力的影响。采用磺酰罗丹明B(SRB)法测定迷迭香酸对乳腺癌MDA-MB-231细胞增殖的影响,Hoechst 33258荧光染色法观察细胞凋亡形态,Annexin VFITC/PI检测细胞凋亡率;同时,细胞划痕实验检测迷迭香酸对MDA-MB-231细胞体外迁移能力的影响,实时荧光PCR(qPCR)法检测Bax、Bcl-2、Caspase-3、MMP-2和MMP-9基因的表达水平。研究结果显示迷迭香酸能抑制乳腺癌MDA-MB-231细胞的增殖,且呈时间剂量依赖性;迷迭香酸处理后的MDA-MB-231细胞出现明显的凋亡形态,且细胞凋亡率明显增加,Bax和caspase-3 mRNA表达水平增加,而Bcl-2 mRNA表达水平降低。另外,迷迭香酸作用后可剂量依赖性地降低MDA-MB-231细胞的体外迁移能力;同时降低MMP-2和MMP-9 mRNA的表达。因此,迷迭香酸能有效的抑制MDA-MB-231细胞的增殖,诱导细胞凋亡,降低细胞迁移能力。

The effects of rosmarinic acid on cell proliferation,apoptosis and migration in breast cancer MDA-MB-231 cells were investigated in this study.After treatment with rosmarinic acid,MDA-MB-231 cell proliferation inhibition rate was examined by SRB assay.Measurement of apoptosis was examined by Hoechst 33258 staining and Annexin V-FITC/PI double staining.Next,scratch wound assay was used to detect the migration activity of MDA-MB-231 cells.Meanwhile,the mRNA levels of Bax,Bcl-2,Caspase-3,MMP-2 and MMP-9 were examined by qRT-PCR.Our results showed that rosmarinic acid inhibited the proliferation of MDA-MB-231 cells in a time-concentration manner.After treatment with rosmarinic acid,MDA-MB-231 cells appeared obvious apoptotic characteristics,and the number of apoptotic cells was significantly increased.Further studies showed that the mRNA levels of Bax and caspase-3 were upregulated,while the mRNA level of Bcl-2 was downregulated in rosmarinic acid-treated cells.In addition,rosmarinic acid blocked the migration of MDA-MB-231 cells in a concentration-dependent manner,which was associated with the downregulation of MMP-9 and MMP-2.Taken together,these results suggested that rosmarinic acid might be a potential drug in human breast cancer treatment by inhibiting proliferation,inducing apoptosis and blocking cell migration.