β-anhydroicaritin Induces Apoptosis of MCF-7 Cells through ROS-Mediated Mitochondrial Apoptosis Pathway
The aim of this study is to investigate the anti-tumor effect of β-anhydroicaritin on human non-small 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 ex-plore the mechanism of anti-tumor effect.MTT assay was used to detect the effects of different concentrations ofβ-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 technol-ogy forecasted β-anhydroicaritin element combination ability with proteins SIRT3 associated with reactive oxy-gen species;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 perme-ability 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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