Chen Jingyao, Zhou Jie, Li Fei, Zhu Yanfeng, Zhang Weiwei, Yu Xiaoping
Department of Public Health, Chengdu Medical College, Chengdu 610500, China.
Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2017 Mar 28;42(3):264-270. doi: 10.11817/j.issn.1672-7347.2017.03.005.
To explore the effect of delphinidin on breast cancer and the underlying mechanisms. Methods: Human epidermal growth factor receptor-2 (HER-2) positive breast cancer cells MDA-MB-453 were treated by delphinidin. Proliferation of MDA-MB-453 cells was detected by CCK-8 after 48 h. TdT-mediated dUTP nick end labeling (TUNEL) assay and Western blot were used to explore apoptotic status for MDA-MB-453 cells. Fluorescence dot assay, immunofluorescence, and Western blot were used to identify autophagy in breast cancer cells. Results: Delphinidin suppressed proliferation of MDA-MB-453 cells. Delphinidin increased the number of TUNEL positive cells. Delphinidin downregulated the expression of caspase-3 and caspase-9, while upregulated the expression of cleaved caspase-3 and cleaved caspase-9 in a dose-dependent manner. Delphinidin enhanced the number of GFP-LC3 punctate dots, LC3 immunofluorescence dots and the expression of LC3-II and ATG5. Delphinidin inhibited the expression of proteins in mTOR signaling pathway, including AKT, mTOR, eIF4E and p70s6k. Conclusion: Delphinidin induced apoptosis and autophagy by inhibition of AKT/mTOR pathway in HER positive breast cancer cells.
探讨飞燕草素对乳腺癌的影响及其潜在机制。方法:用飞燕草素处理人表皮生长因子受体2(HER-2)阳性乳腺癌细胞MDA-MB-453。48小时后通过CCK-8检测MDA-MB-453细胞的增殖情况。采用TdT介导的dUTP缺口末端标记(TUNEL)法和蛋白质免疫印迹法探讨MDA-MB-453细胞的凋亡状态。采用荧光斑点试验、免疫荧光法和蛋白质免疫印迹法鉴定乳腺癌细胞中的自噬情况。结果:飞燕草素抑制MDA-MB-453细胞的增殖。飞燕草素增加了TUNEL阳性细胞的数量。飞燕草素以剂量依赖的方式下调半胱天冬酶-3和半胱天冬酶-9的表达,同时上调裂解的半胱天冬酶-3和裂解的半胱天冬酶-9的表达。飞燕草素增加了绿色荧光蛋白-微管相关蛋白1轻链3(GFP-LC3)点状荧光点、LC3免疫荧光点的数量以及LC3-II和自噬相关蛋白5(ATG5)的表达。飞燕草素抑制了哺乳动物雷帕霉素靶蛋白(mTOR)信号通路中蛋白的表达,包括蛋白激酶B(AKT)、mTOR、真核翻译起始因子4E(eIF4E)和核糖体蛋白S6激酶(p70s6k)。结论:飞燕草素通过抑制HER阳性乳腺癌细胞中的AKT/mTOR通路诱导细胞凋亡和自噬。