Dihydroartemisinin enhances doxorubicin-induced apoptosis of triple negative breast cancer cells by negatively regulating the STAT3/HIF-1α pathway

doi:10.12122/j.issn.1673-4254.2025.02.06

Abstract

Abstract:

Objective To investigate the effects of dihydroartemisinin (DHA) combined with doxorubicin (DOX) on proliferation and apoptosis of triple-negative breast cancer cells and explore the underlying molecular mechanism. Methods MDA-MB-231 cells were treated with 50, 100 or 150 μmol/L DHA, 0.5 μmol/L DOX, or with 50 μmol/L DHA combined with 0.5 μmol/L DOX. The changes in proliferation and survival of the treated cells were examined with MTT assay and colony-forming assay, and cell apoptosis was analyzed with flow cytometry. Western blotting was performed to detect the changes in protein expression levels of PCNA, cleaved PARP, Bcl-2, Bax, STAT3, p-STAT3, HIF-1α and survivin. Results The IC₅₀ of DHA was 131.37±29.87 μmol/L in MDA-MB-231 cells. The cells with the combined treatment with DHA and DOX showed significant suppression of cell proliferation. Treatment with DHA alone induced apoptosis of MDA-MB-231 cells in a dose-dependent manner, but the combined treatment produced a much stronger apoptosis-inducing effect than both DHA and DOX alone. DHA at 150 μmol/L significantly inhibited clone formation of MDA-MB-231 cells, markedly reduced cellular expression levels of PCNA, p-STAT3, HIF-1α and survivin proteins, and obviously increased the expression level of cleaved PARP protein and the Bax/Bcl-2 ratio, and the combined treatment further reduced the expression level of p-STAT3 protein and increased the Bax/Bcl-2 ratio. Conclusion DHA combined with DOX produces significantly enhanced effects for inhibiting cell proliferation and inducing apoptosis in MDA-MB-231 cells possibly as result of DHA-mediated negative regulation of the STAT3/HIF-1α pathway.

Key words: dihydroartemisinin, doxorubicin, STAT3, HIF-1α, triple-negative breast cancer, apoptosis

Di CHEN, Ying LÜ, Yixin GUO, Yirong ZHANG, Ruixuan WANG, Xiaoruo ZHOU, Yuxin CHEN, Xiaohui WU. Dihydroartemisinin enhances doxorubicin-induced apoptosis of triple negative breast cancer cells by negatively regulating the STAT3/HIF-1α pathway[J]. Journal of Southern Medical University, 2025, 45(2): 254-260.

Figures/Tables 8

Fig.1 Effect of dihydroartemisinin (DHA) on proliferation of MDA-MB-231 cells (Mean±SD, n=3).

Fig.2 Effect of DHA on colony formation ability of MDA-MB-231 cells (Mean±SD, n=3). A: Colony-forming assay of the cells. B: Quantitative analysis of the results. **P<0.01 vs 0 μmol/L DHA.

Fig.3 Effect of DHA on apoptosis of MDA-MB-231 cells (Mean±SD, n=3). A: Flow cytometric analysis of cell apoptosis. B: Quantitative analysis of cell apoptosis. **P<0.01, ***P<0.001 vs 0 μmol/L DHA.

Fig.4 Effect of DHA on apoptosis-related protein expressions in MDA-MB-231 cells (Mean±SD, n=3). A: Protein bands in Western blotting. B: Protein expression levels. **P<0.01, ***P<0.001 vs 0 μmol/L DHA.

Fig.5 Effect of DHA on STAT3/HIF-1α signaling pathway protein expressions in MDA-MB-231 cells (Mean±SD, n=3). A: Protein bands in Western blotting. B: Protein expression levels. **P<0.01, ***P<0.001 vs 0 μmol/L DHA.

Fig.6 Effect of DHA combined with DOX on proliferation of MDA-MB-231 cells (Mean±SD, n=3). **P<0.01, ***P<0.001 vs DHA or DOX.

Fig.7 Effect of DHA combined with DOX on apoptosis of MDA-MB-231 cells (Mean±SD, n=3). A: Flow cytometric analysis of cell apoptosis. B: Quantitative analysis of cell apoptosis. ***P<0.01 vs DOX or DHA.

Fig.8 Effect of DHA and DOX on STAT3/HIF-1α signaling pathway protein expressions in MDA-MB-231 cells (Mean±SD, n=3). A: Protein bands in Western blotting. B: Protein expression levels. *P<0.05,**P<0.01, ***P<0.001 vs Control or DOX.

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