蒲公英根水提物诱导MDA-MB-231细胞凋亡及其作用机制研究

Induction of apoptosis and mechanism of MDA-MB-231 cells by water extract of dandelion root

目的探究蒲公英根水提物对人三阴性乳腺癌细胞MDA-MB-231增殖和凋亡的影响及其作用机制。方法采用Alamar Blue法检测蒲公英根水提物的细胞毒性,以及对MDA-MB-231细胞增殖的抑制作用;通过形态学观察、DAPI染色、流式细胞术,检测蒲公英根水提物对MDA-MB-231细胞凋亡的影响;Western blot分析蒲公英根水提物诱导MDA-MB-231细胞凋亡的作用机制。结果蒲公英各部位提取物无细胞毒性;且7.5、15 g·L^(-1)的蒲公英根水提物明显抑制MDA-MB-231细胞的增殖,细胞出现明显的凋亡特征,流式细胞术进一步证实了蒲公英根水提物能够诱导MDA-MB-231细胞凋亡;蒲公英根水提物呈浓度依赖性降低Bcl-2的蛋白表达,增加Bax、p53、活性c-PARP、caspase-8、caspase-9和caspase-3表达量; caspase-8抑制剂(Z-IETD-FMK)和caspas-9抑制剂(Z-LEHD-FMK)能够逆转蒲公英水提物对MDA-MB-231细胞的增殖抑制活性。结论蒲公英根水提物对人三阴性乳腺癌细胞MDA-MB-231具有明显的抑制作用,对治疗三阴性乳腺癌具有潜在的临床应用价值。

Aim To discuss the effects of water extract of dandelion root on the proliferation and apoptosis of human triple negative breast cancer cell line MDA-MB-231 and its mechanism.Methods In this study,the cytotoxicity of dandelion root extract and the inhibition of MDA-MB-231 cell proliferation were detected by Alamar Blue method.Effects of water extracts of dandelion root on apoptosis of MDA-MB-231 cells were assessed by morphological observation,DAPI staining and flow cytometry;The mechanism of dandelion root water extract induced apoptosis of MDA-MB-231 cells was analysed by Western blot.Results Extracts from various parts of dandelion were not cytotoxic;furthermore,7.5 and 15 g·L^-1 water extract of dandelion significantly inhibited the proliferation of MDA-MB-231 cells,and the cells showed obvious apoptotic characteristics.Flow cytometry further confirmed the aqueous extract of dandelion root could induce the apoptosis of MDA-MB-231 cells.In terms of molecular mechanism,the water extract of dandelion root decreased the expression of Bcl-2 protein in a concentration-dependent manner,while the expressions of Bax,p53,active c-PARP,caspase-8,caspase-9 and caspase-3 increased.Caspase-8 inhibitor(Z-IETD-FMK)and caspase-9 inhibitor(Z-LEHD-FMK)were able to reverse the proliferation inhibitory activity of dandelion water extract on MDA-MB-231 cells.Conclusions The water extract of dandelion has a significant inhibitory effect on human triple negative breast cancer MDA-MB-231 cells,pointing to its potential clinical application value for the treatment of triple negative breast cancer.