Effect of Digoxin on apoptosis of MDA-MB-231 cells in triple negative breast cancer
LYU Yuchen1 WU Miaomiao2 FANG Kun2 CHEN Wenxu1 SHEN Aizong2 LIU Tongzhu2 WANG Ning1
1.College of Pharmacy, Anhui University of Chinese Medicine, Anhui Province, Hefei 230012, China;
2.Biomedical Engineering Agency, the First Affiliated Hospital of USTC Anhui Provincial Hospital, Anhui Province, Hefei 230001, China
Abstract：Objective To investigate the effect of Digoxin on apoptosis of MDA-MB-231 cells of triple negative breast cancer in vitro, and to reveal its possible mechanism. Methods Breast cancer MDA-MB-231 cells were cultured in vitro and divided into control group and drug administration group (25, 50, 100, 200 nmol/L Digoxin). CCK-8 method was used to detect the effects of Digoxin at different concentrations on the activity of MDA-MB-231 cells at different action time (24, 48, 72 h). The effects of Digoxin at different concentrations on apoptosis and mitochondrial membrane potential were quantitatively investigated by flow cytometry. Expression of Bax, Bcl-2, P62 and LC3B in MDA-MB-231 cells was detected by Western blot. The effects of Digoxin at different concentrations on autophagosomes in MDA-MB-231 cells were observed by transmission electron microscopy. Results CCK-8 results indicated that, compared with the control group, the activity of MDA-MB-231 cells decreased in the drug administration group at different time, with statistically significant differences (P ＜ 0.05 or P ＜ 0.01). Annexin V/PI double staining results indicated that compared with the control group, the apoptosis rate of the treated group increased with a statistically significant difference (P ＜ 0.05 or P ＜ 0.01). The results of JC-1 staining indicated that compared with the control group, 50, 100 and 200 nmol/L of Digoxin reduced the mitochondrial membrane potential of MDA-MB-231 cells, and the proportion of JC-1 decreased, with statistically significant differences (P ＜ 0.05 or P ＜ 0.01). The results of fluoroscopy indicated that, compared with the control group, the cell morphology of the drug administration group underwent apoptotic changes, and the formation of autophagosomes was observed in the drug administration group. Western blot results indicated that, compared with the control group, the administration group up-regulated the expression of pro-apoptotic factor Bax, down-regulated the expression of anti-apoptotic factor Bcl-2, and up-regulated the expression of autophagy marker protein LC3B, with statistically significant differences (P ＜ 0.05 or P ＜ 0.01); compared with the control group, 50, 100 and 200 nmol/L of Digoxin decreased the expression of P62, with statistically significant differences (P < 0.05 or P < 0.01). Conclusion Digoxin inhibits the activity of MDA-MB-231 cells, promotes apoptosis, induces autophagy, and may have a potential effect on triple negative breast cancer.
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2019, Vol. 16 
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