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Phenformin Induces Cell Cycle Change, Apoptosis, and Mesenchymal-Epithelial Transition and Regulates the AMPK/mTOR/p70s6k and MAPK/ERK Pathways in Breast Cancer Cells.

Liu Z, Ren L, Liu C, Xia T, Zha X, Wang S - PLoS ONE (2015)

Bottom Line: Emerging evidence has demonstrated the potent anti-tumor effect of biguanides, among which phenformin was reported to potentially be a more active anti-cancer agent than metformin.Furthermore, our results suggest that phenformin inhibits breast cancer cell metastasis after intracardiac injection into nude mice.Taken together, our study further confirms the potential benefit of phenformin in breast cancer treatment and provides novel mechanistic insight into its anti-cancer activity in breast cancer.

View Article: PubMed Central - PubMed

Affiliation: Department of Breast Surgery, the First Affiliated Hospital of Nanjing Medical University, Nanjing, China.

ABSTRACT
Breast cancer remains a world-wide challenge, and additional anti-cancer therapies are still urgently needed. Emerging evidence has demonstrated the potent anti-tumor effect of biguanides, among which phenformin was reported to potentially be a more active anti-cancer agent than metformin. However, little attention has been given to the role of phenformin in breast cancer. In this study, we reveal the role of phenformin in cell death of the MCF7, ZR-75-1, MDA-MB-231 and SUM1315 breast cancer cell lines. The respective IC50 values of phenformin in MCF7, ZR-75-1, MDA-MB-231 and SUM1315 cells were 1.184±0.045 mM, 0.665±0.007 mM, 2.347±0.010 mM and 1.885±0.015 mM (mean± standard error). Phenformin induced cell cycle change and apoptosis in breast cancer cells via the AMPK/mTOR/p70s6k and MAPK/ERK pathways. Interestingly, phenformin induced MET (mesenchymal-epithelial transition) and decreased the migration rate in breast cancer cell lines. Furthermore, our results suggest that phenformin inhibits breast cancer cell metastasis after intracardiac injection into nude mice. Taken together, our study further confirms the potential benefit of phenformin in breast cancer treatment and provides novel mechanistic insight into its anti-cancer activity in breast cancer.

No MeSH data available.


Related in: MedlinePlus

Phenformin downregulates cyclin D1 expression and inactivates the MAPK/ERK pathway.(A) MCF7, ZR-75-1, MDA-MB-231 and SUM1315 cells were treated with or without phenformin for 24 hours. Cell extracts were analyzed by western blotting to detect the expression of cyclin D1, p-ERK, ERK and GAPDH. (B) Western blotting ratio analysis of p-ERK to ERK, (C) Cyclin D1 to GAPDH. The data are presented as the mean±SEM of three replicates per group. Asterisks indicate significant differences at p<0.05 by Student’s t test. Phenformin-treated cells were labeled as P and control cells were labeled as C.
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pone.0131207.g003: Phenformin downregulates cyclin D1 expression and inactivates the MAPK/ERK pathway.(A) MCF7, ZR-75-1, MDA-MB-231 and SUM1315 cells were treated with or without phenformin for 24 hours. Cell extracts were analyzed by western blotting to detect the expression of cyclin D1, p-ERK, ERK and GAPDH. (B) Western blotting ratio analysis of p-ERK to ERK, (C) Cyclin D1 to GAPDH. The data are presented as the mean±SEM of three replicates per group. Asterisks indicate significant differences at p<0.05 by Student’s t test. Phenformin-treated cells were labeled as P and control cells were labeled as C.

Mentions: To examine whether the growth inhibitory effect of phenformin on breast cancer cells was partly due to cell cycle change, we performed cell cycle analysis using PI staining. MCF7, ZR-75-1, MDA-MB-231 and SUM1315 cells were exposed to 1.184 mM, 0.665 mM, 2.347 mM, and 1.885 mM phenformin, (the IC50 of each cell line) respectively for 24 hours. The percentage of phenformin-treated MCF7, ZR-75-1, MDA-MB-231 and SUM1315 cells in G1 significantly increased to 67.7% (p = 0.01), 35.9% (p = 0.0088), 29.5% (p = 0.0058) and 54.4% (p = 0.0168), respectively (Fig 2). Consistent with the CCK8 assay results, the lowest percentage of cells in G1 was observed in MDA-MB-231 cells. To further confirm the increased cells in G1 induced by phenformin, we performed western blotting assays to detect cyclin D1 expression. Notably, significantly lower levels of cyclin D1 protein were observed in all four breast cancer cell lines treated with phenformin (Fig 3), which suggested that phenformin might regulate the cell cycle of breast cancer cells by modulating cyclin D1. Interestingly, the regulation of ERK phosphorylation by phenformin diverged between the ERα-positive and ERα-negative breast cancer cell lines. The p-ERK levels in MCF7 and ZR-75-1 cells decreased significantly upon phenformin treatment. In contrast, p-ERK levels did not change in MDA-MB-231 or SUM1315 cells after treatment with phenformin (Fig 3).


Phenformin Induces Cell Cycle Change, Apoptosis, and Mesenchymal-Epithelial Transition and Regulates the AMPK/mTOR/p70s6k and MAPK/ERK Pathways in Breast Cancer Cells.

Liu Z, Ren L, Liu C, Xia T, Zha X, Wang S - PLoS ONE (2015)

Phenformin downregulates cyclin D1 expression and inactivates the MAPK/ERK pathway.(A) MCF7, ZR-75-1, MDA-MB-231 and SUM1315 cells were treated with or without phenformin for 24 hours. Cell extracts were analyzed by western blotting to detect the expression of cyclin D1, p-ERK, ERK and GAPDH. (B) Western blotting ratio analysis of p-ERK to ERK, (C) Cyclin D1 to GAPDH. The data are presented as the mean±SEM of three replicates per group. Asterisks indicate significant differences at p<0.05 by Student’s t test. Phenformin-treated cells were labeled as P and control cells were labeled as C.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4482683&req=5

pone.0131207.g003: Phenformin downregulates cyclin D1 expression and inactivates the MAPK/ERK pathway.(A) MCF7, ZR-75-1, MDA-MB-231 and SUM1315 cells were treated with or without phenformin for 24 hours. Cell extracts were analyzed by western blotting to detect the expression of cyclin D1, p-ERK, ERK and GAPDH. (B) Western blotting ratio analysis of p-ERK to ERK, (C) Cyclin D1 to GAPDH. The data are presented as the mean±SEM of three replicates per group. Asterisks indicate significant differences at p<0.05 by Student’s t test. Phenformin-treated cells were labeled as P and control cells were labeled as C.
Mentions: To examine whether the growth inhibitory effect of phenformin on breast cancer cells was partly due to cell cycle change, we performed cell cycle analysis using PI staining. MCF7, ZR-75-1, MDA-MB-231 and SUM1315 cells were exposed to 1.184 mM, 0.665 mM, 2.347 mM, and 1.885 mM phenformin, (the IC50 of each cell line) respectively for 24 hours. The percentage of phenformin-treated MCF7, ZR-75-1, MDA-MB-231 and SUM1315 cells in G1 significantly increased to 67.7% (p = 0.01), 35.9% (p = 0.0088), 29.5% (p = 0.0058) and 54.4% (p = 0.0168), respectively (Fig 2). Consistent with the CCK8 assay results, the lowest percentage of cells in G1 was observed in MDA-MB-231 cells. To further confirm the increased cells in G1 induced by phenformin, we performed western blotting assays to detect cyclin D1 expression. Notably, significantly lower levels of cyclin D1 protein were observed in all four breast cancer cell lines treated with phenformin (Fig 3), which suggested that phenformin might regulate the cell cycle of breast cancer cells by modulating cyclin D1. Interestingly, the regulation of ERK phosphorylation by phenformin diverged between the ERα-positive and ERα-negative breast cancer cell lines. The p-ERK levels in MCF7 and ZR-75-1 cells decreased significantly upon phenformin treatment. In contrast, p-ERK levels did not change in MDA-MB-231 or SUM1315 cells after treatment with phenformin (Fig 3).

Bottom Line: Emerging evidence has demonstrated the potent anti-tumor effect of biguanides, among which phenformin was reported to potentially be a more active anti-cancer agent than metformin.Furthermore, our results suggest that phenformin inhibits breast cancer cell metastasis after intracardiac injection into nude mice.Taken together, our study further confirms the potential benefit of phenformin in breast cancer treatment and provides novel mechanistic insight into its anti-cancer activity in breast cancer.

View Article: PubMed Central - PubMed

Affiliation: Department of Breast Surgery, the First Affiliated Hospital of Nanjing Medical University, Nanjing, China.

ABSTRACT
Breast cancer remains a world-wide challenge, and additional anti-cancer therapies are still urgently needed. Emerging evidence has demonstrated the potent anti-tumor effect of biguanides, among which phenformin was reported to potentially be a more active anti-cancer agent than metformin. However, little attention has been given to the role of phenformin in breast cancer. In this study, we reveal the role of phenformin in cell death of the MCF7, ZR-75-1, MDA-MB-231 and SUM1315 breast cancer cell lines. The respective IC50 values of phenformin in MCF7, ZR-75-1, MDA-MB-231 and SUM1315 cells were 1.184±0.045 mM, 0.665±0.007 mM, 2.347±0.010 mM and 1.885±0.015 mM (mean± standard error). Phenformin induced cell cycle change and apoptosis in breast cancer cells via the AMPK/mTOR/p70s6k and MAPK/ERK pathways. Interestingly, phenformin induced MET (mesenchymal-epithelial transition) and decreased the migration rate in breast cancer cell lines. Furthermore, our results suggest that phenformin inhibits breast cancer cell metastasis after intracardiac injection into nude mice. Taken together, our study further confirms the potential benefit of phenformin in breast cancer treatment and provides novel mechanistic insight into its anti-cancer activity in breast cancer.

No MeSH data available.


Related in: MedlinePlus