Long-Term Glucose Restriction with or without β-Hydroxybutyrate Enrichment Distinctively Alters Epithelial-Mesenchymal Transition-Related Signalings in Ovarian Cancer Cells

Abstract

The beneficial impacts of the ketogenic diet and metabolic reprograming were recently reported for ovarian cancer patients. In this study, the effects of glucose restriction with or without beta-hydroxybutyrate (bHB) enrichment were studied in drug-resistant CD133high A2780CP and CD133low SK-OV-3 ovarian cancer cells to scrutinize the impact of experimental ketosis on ATP production, epithelial to mesenchymal transition (EMT), and related signaling pathways including Wnt, Hippo, and Hedgehog. Cells were adapted and maintained for a month with restricted levels of glucose (250 mg/l) with or without the therapeutic concentration of bHB (5 mM). Quantitative PCR, Western blot analysis, flow cytometry, chemiluminescence, and wound healing assay were used in this study. Glucose restriction and bHB enrichment reduced the stemness marker and diminished In Vitro migration in both cell lines. Glucose restriction significantly reduced ATP levels in both cells, but bHB enrichment was partially compensated for the ATP levels solely in SK-OV-3 cells. Glucose restriction mainly inhibited the Wnt pathway in the CD133high A2780CP cells, but the Hedgehog pathway was the main target in CD133low SK-OV-3 cells. In Conclusion, Prior targeted evaluations of key genes' expression would help to predict the distinctive impacts of metabolic fuels and to optimize the efficacy of ketogenic diets.

Acknowledgments

A part of the article has been extracted from the thesis written by Hossein Ghahremani in the School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran (registration number: (180).

Disclosure Statement

The authors declare that there is no conflict of interest regarding the publication of this article.

Additional information

Funding

The study was approved by the local Committee for Ethics with reference numbers IR.SBMU. MSP.REC.1396.538 and IR.SBMU.SM.REC.1395.571. Part of this study was supported by a research (grant no. 9803-66009803) from the deputy of research, Shahid Beheshti University of Medical Sciences.

Notes on contributors

Hossein Ghahremani

Siamak Salami, Majid Sirati-Sabet, and Hossein Ghahremani designed the study. Siamak Salami and Majid Sirati-Sabet supervised the whole project. Hossein Ghahremani, Saeedeh Nabati, Hanieh Tahmori, Tahmineh Peirouvi, Majid Sirati-Sabet and Siamak Salami were substantially involved in the experiments. Siamak Salami, Majid Sirati-Sabet, Hossein Ghahremani, and Tahmineh Peirouvi were contributed to technical and statistical analyses and interpretation of the results. Siamak Salami, Majid Sirati-Sabet and Hossein Ghahremani drafted the manuscript and all authors checked and approved the final manuscript.

Saeedeh Nabati

Siamak Salami, Majid Sirati-Sabet, and Hossein Ghahremani designed the study. Siamak Salami and Majid Sirati-Sabet supervised the whole project. Hossein Ghahremani, Saeedeh Nabati, Hanieh Tahmori, Tahmineh Peirouvi, Majid Sirati-Sabet and Siamak Salami were substantially involved in the experiments. Siamak Salami, Majid Sirati-Sabet, Hossein Ghahremani, and Tahmineh Peirouvi were contributed to technical and statistical analyses and interpretation of the results. Siamak Salami, Majid Sirati-Sabet and Hossein Ghahremani drafted the manuscript and all authors checked and approved the final manuscript.

Hanieh Tahmori

Siamak Salami, Majid Sirati-Sabet, and Hossein Ghahremani designed the study. Siamak Salami and Majid Sirati-Sabet supervised the whole project. Hossein Ghahremani, Saeedeh Nabati, Hanieh Tahmori, Tahmineh Peirouvi, Majid Sirati-Sabet and Siamak Salami were substantially involved in the experiments. Siamak Salami, Majid Sirati-Sabet, Hossein Ghahremani, and Tahmineh Peirouvi were contributed to technical and statistical analyses and interpretation of the results. Siamak Salami, Majid Sirati-Sabet and Hossein Ghahremani drafted the manuscript and all authors checked and approved the final manuscript.

Tahmineh Peirouvi

Siamak Salami, Majid Sirati-Sabet, and Hossein Ghahremani designed the study. Siamak Salami and Majid Sirati-Sabet supervised the whole project. Hossein Ghahremani, Saeedeh Nabati, Hanieh Tahmori, Tahmineh Peirouvi, Majid Sirati-Sabet and Siamak Salami were substantially involved in the experiments. Siamak Salami, Majid Sirati-Sabet, Hossein Ghahremani, and Tahmineh Peirouvi were contributed to technical and statistical analyses and interpretation of the results. Siamak Salami, Majid Sirati-Sabet and Hossein Ghahremani drafted the manuscript and all authors checked and approved the final manuscript.

Majid Sirati-Sabet

Siamak Salami, Majid Sirati-Sabet, and Hossein Ghahremani designed the study. Siamak Salami and Majid Sirati-Sabet supervised the whole project. Hossein Ghahremani, Saeedeh Nabati, Hanieh Tahmori, Tahmineh Peirouvi, Majid Sirati-Sabet and Siamak Salami were substantially involved in the experiments. Siamak Salami, Majid Sirati-Sabet, Hossein Ghahremani, and Tahmineh Peirouvi were contributed to technical and statistical analyses and interpretation of the results. Siamak Salami, Majid Sirati-Sabet and Hossein Ghahremani drafted the manuscript and all authors checked and approved the final manuscript.

Siamak Salami

Siamak Salami, Majid Sirati-Sabet, and Hossein Ghahremani designed the study. Siamak Salami and Majid Sirati-Sabet supervised the whole project. Hossein Ghahremani, Saeedeh Nabati, Hanieh Tahmori, Tahmineh Peirouvi, Majid Sirati-Sabet and Siamak Salami were substantially involved in the experiments. Siamak Salami, Majid Sirati-Sabet, Hossein Ghahremani, and Tahmineh Peirouvi were contributed to technical and statistical analyses and interpretation of the results. Siamak Salami, Majid Sirati-Sabet and Hossein Ghahremani drafted the manuscript and all authors checked and approved the final manuscript.