总丹参酮通过线粒体途径诱导乳腺癌细胞MCF-7凋亡的实验研究  

Study on Apoptosis of Breast Cancer Cell MCF-7 Induced by Total Tanshinone Through Mitochondrial Pathway

作　　者：李虎业 孔德元 窦增花 郑科 LI Hu-ye;KONG De-yuan;DOU Zeng-hua;ZHENG Ke(The Fourth People's Hospital of Qinghai Province, Xining 810000, China;Surgical Laboratory, The Affiliated Hospital of Chongqing Medical University,Chongqing 400016,China)

机构地区：[1]青海省第四人民医院,青海西宁810000 [2]重庆医科大学附属第一医院外科实验室,重庆400016

出　　处：《中药材》2019年第1期184-188,共5页Journal of Chinese Medicinal Materials

基　　金：国家自然科学基金(81202090)

摘　　要：目的:探讨总丹参酮对乳腺癌细胞MCF-7凋亡的影响及作用机制。方法:体外培养乳腺癌细胞MCF-7,随机分为4组,加入不同浓度总丹参酮(0、0.5、1、2μg/mL)处理48 h。四甲基偶氮唑(MTT)法检测各组细胞的增殖抑制率,流式细胞术检测细胞的凋亡率,免疫印迹试验(Western blot)检测细胞浆中活化的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved Caspase-3)、活化的含半胱氨酸的天冬氨酸蛋白水解酶9(Cleaved Caspase-9)、活化的多聚ADP-核糖聚合酶(Cleaved PARP)、Smac、Cytochrome c、和线粒体Bcl-2相关X蛋白(Bax)、Bak蛋白的表达,采用试剂盒检测Caspase-3、Caspase-9的活性。结果:总丹参酮能显著抑制乳腺癌细胞MCF-7的活力,诱导细胞凋亡,显著上调Cleaved PARP、Cleaved Caspase-3、Cleaved Caspase-9、Smac、Cytochrome c蛋白表达及线粒体中Bax、Bak蛋白的表达(P<0.05),而细胞中Bax、Bak蛋白水平无明显变化。结论:总丹参酮能显著抑制乳腺癌细胞MCF-7活力,诱导凋亡,其作用机制与线粒体途径有关。Objective:To investigate the effect of total tanshinone(TTN)on the apoptosis of breast cancer cells MCF-7 and its mechanism.Methods:Breast cancer cells MCF-7 were cultured in vitro and randomly divided into four groups with added different concentrations of TTN(0,0.5,1,2 μg/mL)for 48 h.The proliferation inhibition rates of cells in each group were detected by MTT assay and the apoptosis rate was measured by flow cytometry.The protein levels of cleaved cysteinyl aspartate specific proteinase 3(Cleaved Caspase-3),cleaved cysteinyl aspartate specific proteinase 9(Cleaved Caspase-9),cleaved poly ADP-ribose polymerase(Cleaved PARP),Smac,Cytochrome c and Bcl-2 related X protein(Bax),Bak protein in mitochondria were detected by Western blot.The activities of Caspase-3 and Caspase-9 were detected by kit.Results:TTN could significantly inhibite the activity of MCF-7 cells,induce apoptosis and significantly increase the expression levels of Cleaved PARP,Cleaved Caspase-3,Cleaved Caspase-9,Smac,Cytochrome c and Bax and Bak in mitochondria(P<0.05),while the levels of Bax and Bak proteins in cells had no significant changes.Conclusion:TTN can significantly inhibit MCF-7 activity and induce apoptosis in breast cancer cells,and the mechanism is related to mitochondrial pathway.

关 键 词：总丹参酮 乳腺癌 凋亡 线粒体途径 

分 类 号：R285.5[医药卫生—中药学]