β-anhydroicaritin Induces Apoptosis of MCF-7 Cells through ROS-Mediated Mitochondrial Apoptosis Pathway
SHU Hui1, ZHAO Yan1, LIU Fangfei1, HAN Jiahong1, CAI Enbo1*, QI Wenqian2*
The aim of this study is to investigate the anti-tumor effect of β-anhydroicaritin on human nonsmall cell lung cancer A549, breast cancer MCF-7, prostate cancer PC-3M, cervical squamous cell carcinoma SiHa and pancreatic cancer PANC-1 cells, and to screen out the cell lines with the best inhibitory activity to explore the mechanism of anti-tumor effect. MTT assay was used to detect the effects of different concentrations of β-anhydroicaritin on the proliferation of five kinds of tumor cells, and the breast cancer MCF-7 cells with more prominent inhibitory effect of β-anhydroicaritin were selected for subsequent experiments based on IC50; DAPI staining and flow cytometry were used to detect cell apoptosis; DCFH-DA method was used to detect the level of reactive oxygen species in cells; the opening of mitochondrial permeability transition pore was detected by using Calcein AM probe; JC-1 probe was used to detect mitochondrial membrane potential; molecular docking technology forecasted β-anhydroicaritin element combination ability with proteins SIRT3 associated with reactive oxygenspecies; Western blot was used to detect the expression levels of SIRT3, Cytochorme C, Cleaved caspase-3,Cleaved caspase-9, PARP, and Cleaved PARP in the cells. The results showed that β-anhydroicaritin could stimulate the level of reactive oxygen species in tumor cells by regulating the expression level of SIRT3 protein, which leads to mitochondrial dysfunction in MCF-7 cells, and then stimulated the abnormal opening of mitochondrial permeability transition pore, greatly reduced the membrane potential, and finally induced apoptosis of MCF-7 cells. These effects may be mediated by ROS (reactive oxygen species)-mediated mitochondrial apoptosis.
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