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

Ku JM, Kim SR, Hong SH, Choi HS, Seo HS, Shin YC, Ko SG - Mol. Cell. Biochem. (2015)

Bottom Line: Additionally, cucurbitacin D led to an increase in the IκBα level in the cytosol and a decrease in the p-NF-κB level in the nucleus.Finally, cucurbitacin D inhibited translocation of Stat3 and NF-κB and decreased transcriptional activity in the nucleus.Cucurbitacin D could be used as a useful compound to treat adriamycin-resistant patients.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Clinical Biology and Pharmacogenomics, Department of Preventive Medicine, College of Oriental Medicine, Kyung Hee University, Seoul, 130-701, Republic of Korea.

ABSTRACT
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-κB, IκB, and caspases were measured by Western blot analysis. Nuclear staining of Stat3 and NF-κB was measured by immunocytochemistry. STAT3 and NF-κB transcriptional activity was detected by STAT3 and NF-κB 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 IκBα level in the cytosol and a decrease in the p-NF-κB level in the nucleus. Finally, cucurbitacin D inhibited translocation of Stat3 and NF-κB and decreased transcriptional activity in the nucleus. Cucurbitacin D decreases cell proliferation and induces apoptosis by inhibiting Stat3 and NF-κB signaling in doxorubicin-resistant breast cancer cells. Cucurbitacin D could be used as a useful compound to treat adriamycin-resistant patients.

No MeSH data available.


Related in: MedlinePlus

Cucurbitacin D suppresses p-Stat3 expression in MCF7/ADR Cells. Constitutive activation of Stat3 was detected in MCF7/ADR cells. p-STAT3 was strongly expressed in MCF7/ADR cells. a Doxorubicin increases constitutive STAT3 phosphorylation in a time-dependent manner in MCF7 cells. b Cucurbitacin D inhibits p-Stat3 expression in MCF7/ADR cells. c Whole-cell lysates were analyzed by Western blot with anti-pStat3, anti-Stat3, and anti-tubulin antibodies
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Fig2: Cucurbitacin D suppresses p-Stat3 expression in MCF7/ADR Cells. Constitutive activation of Stat3 was detected in MCF7/ADR cells. p-STAT3 was strongly expressed in MCF7/ADR cells. a Doxorubicin increases constitutive STAT3 phosphorylation in a time-dependent manner in MCF7 cells. b Cucurbitacin D inhibits p-Stat3 expression in MCF7/ADR cells. c Whole-cell lysates were analyzed by Western blot with anti-pStat3, anti-Stat3, and anti-tubulin antibodies

Mentions: To determine the relationship between Stat3 and doxorubicin resistance in human breast cancer cells, we analyzed the expression of p-STAT3 and Stat3 in MCF7/ADR cells and MCF7 cells. We found that p-Stat3 expression was significantly overexpressed in MCF7/ADR cells than in the MCF7 cell line (Fig. 2a). Next, we determined whether doxorubicin increased p-Stat3 expression in MCF7 cells. We found that doxorubicin alone increased p-STAT3 level in a time-dependent manner (Fig. 2b).Fig. 2


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

Ku JM, Kim SR, Hong SH, Choi HS, Seo HS, Shin YC, Ko SG - Mol. Cell. Biochem. (2015)

Cucurbitacin D suppresses p-Stat3 expression in MCF7/ADR Cells. Constitutive activation of Stat3 was detected in MCF7/ADR cells. p-STAT3 was strongly expressed in MCF7/ADR cells. a Doxorubicin increases constitutive STAT3 phosphorylation in a time-dependent manner in MCF7 cells. b Cucurbitacin D inhibits p-Stat3 expression in MCF7/ADR cells. c Whole-cell lysates were analyzed by Western blot with anti-pStat3, anti-Stat3, and anti-tubulin antibodies
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4589559&req=5

Fig2: Cucurbitacin D suppresses p-Stat3 expression in MCF7/ADR Cells. Constitutive activation of Stat3 was detected in MCF7/ADR cells. p-STAT3 was strongly expressed in MCF7/ADR cells. a Doxorubicin increases constitutive STAT3 phosphorylation in a time-dependent manner in MCF7 cells. b Cucurbitacin D inhibits p-Stat3 expression in MCF7/ADR cells. c Whole-cell lysates were analyzed by Western blot with anti-pStat3, anti-Stat3, and anti-tubulin antibodies
Mentions: To determine the relationship between Stat3 and doxorubicin resistance in human breast cancer cells, we analyzed the expression of p-STAT3 and Stat3 in MCF7/ADR cells and MCF7 cells. We found that p-Stat3 expression was significantly overexpressed in MCF7/ADR cells than in the MCF7 cell line (Fig. 2a). Next, we determined whether doxorubicin increased p-Stat3 expression in MCF7 cells. We found that doxorubicin alone increased p-STAT3 level in a time-dependent manner (Fig. 2b).Fig. 2

Bottom Line: Additionally, cucurbitacin D led to an increase in the IκBα level in the cytosol and a decrease in the p-NF-κB level in the nucleus.Finally, cucurbitacin D inhibited translocation of Stat3 and NF-κB and decreased transcriptional activity in the nucleus.Cucurbitacin D could be used as a useful compound to treat adriamycin-resistant patients.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Clinical Biology and Pharmacogenomics, Department of Preventive Medicine, College of Oriental Medicine, Kyung Hee University, Seoul, 130-701, Republic of Korea.

ABSTRACT
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-κB, IκB, and caspases were measured by Western blot analysis. Nuclear staining of Stat3 and NF-κB was measured by immunocytochemistry. STAT3 and NF-κB transcriptional activity was detected by STAT3 and NF-κB 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 IκBα level in the cytosol and a decrease in the p-NF-κB level in the nucleus. Finally, cucurbitacin D inhibited translocation of Stat3 and NF-κB and decreased transcriptional activity in the nucleus. Cucurbitacin D decreases cell proliferation and induces apoptosis by inhibiting Stat3 and NF-κB signaling in doxorubicin-resistant breast cancer cells. Cucurbitacin D could be used as a useful compound to treat adriamycin-resistant patients.

No MeSH data available.


Related in: MedlinePlus