Cucurbitacin D induces cell cycle arrest and apoptosis by inhibiting STAT3 and NF-kappa B signaling in doxorubicin-resistant human breast carcinoma (MCF7/ADR) cells

摘要

Breast cancer is the most common cancer for women and is a major cause of mortality in women. Doxorubicin is a generally used chemotherapy drug for breast cancer. However, multidrug resistance of breast cancer interferes with the chemotherapy. We examined whether cucurbitacin D affects doxorubicin resistance of MCF7/ADR breast cancer cells. Cell viability was measured by MTT assay. Levels of p-STAT3, p-NF-kappa B, I kappa B, and caspases were measured by Western blot analysis. Nuclear staining of Stat3 and NF-kappa B was measured by immunocytochemistry. STAT3 and NF-kappa B transcriptional activity was detected by STAT3 and NF-kappa B luciferase reporter gene assays. Analysis of cell cycle arrest was performed by flow cytometry. Induction of apoptosis by cucurbitacin D was measured by Annexin V-FITC/propidium iodide assay. More than 90 % of MCF7/ADR cells lived upon treatment with doxorubicin for 24 h. However, upon treatment with cucurbitacin D, cell death was more than 60 %. Co-administration of cucurbitacin D and doxorubicin induced apoptosis, and G2/M cell cycle arrest, and inhibited upregulated Stat3 by doxorubicin on MCF7/ADR cells. Additionally, cucurbitacin D led to an increase in the I kappa B alpha level in the cytosol and a decrease in the p-NF-kappa B level in the nucleus. Finally, cucurbitacin D inhibited translocation of Stat3 and NF-kappa B and decreased transcriptional activity in the nucleus. Cucurbitacin D decreases cell proliferation and induces apoptosis by inhibiting Stat3 and NF-kappa B signaling in doxorubicin-resistant breast cancer cells. Cucurbitacin D could be used as a useful compound to treat adriamycin-resistant patients.