Abstract
The effects of insulin on the doxorubicin (Dox) sensitivity of breast cancer cell line MCF-7 and its Dox-resistant counterpart MCF-7/Dox were studied and glucose metabolism, content of essential minerals, and the expression of several microRNAs in these cells upon exposure to insulin and Dox were compared. Cell viability colorimetric assay, colorimetric enzymatic technique, flow cytometry, immunocytochemical techniques, inductively-coupled plasma atomic emission spectroscopy, and quantitative polymerase chain reaction were used in the study. We found that insulin in high concentration significantly suppressed Dox toxicity, especially in parental MCF-7 cell line. The increase in proliferative activity triggered by insulin in MCF-7 but not MCF-7/Dox cells occurred in the setting of the increased level of specific binding sites for insulin and increased glucose uptake. Insulin treatment of MCF-7 cells in low and high concentrations resulted in the increase of Mg, Ca, and Zn content while in DOX-resistant cells, only Mg content increased upon exposure to insulin. High concentration of insulin increased the expression of kinase Akt1, P-glycoprotein 1 (P-gp1) and DNA excision repair protein ERCC-1 in MCF-7 cells, while in MCF-7/Dox cells, Akt1 expression decreased, and cytoplasmic expression of P-gp1 increased. In addition, insulin treatment affected expression of miR-122-5p, miR-133a-3p, miR-200b-3p, and miR-320a-3p. The decreased manifestation of biological effects of insulin in Dox-resistant cells could be partly explained by the different patterns of energy metabolism in MCF-7 cells and their Dox-resistant counterpart.
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Abbreviations
- BC:
-
Breast cancer
- Dox:
-
Doxorubicin
- ER:
-
Estrogen receptor
- ERCC-1:
-
DNA excision repair protein
- FITC:
-
Fluorescein isothiocyanate
- IGF-1:
-
Insulin-like growth factor 1
- miR:
-
microRNA
- P-gp1:
-
P-glycoprotein 1
- RT-qPCR:
-
Reverse transcription quantitative real-time polymerase chain reaction
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Acknowledgements
The authors would like to thank Dr. N.N. Khranovskaya for assisting in flow cytometry work.
Funding
The work was supported by scientific grant No. 0117U002034 “Molecular and Biological Factors of Malignant Cells Heterogeneity and Variability of Clinical Course of Hormone Dependent Tumors” to Dr. Vasyl Chekhun from the target research program of the National Academy of Sciences of Ukraine “Molecular Genetics and Biochemical Mechanisms of Regulation of Cellular and Systemic Interactions in Physiological and Pathological Conditions.”
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O.L., M.Z., T.K., and N.V.: investigation and validation; T.B., N.L., Y.L., and I.A.: methodology and investigation; A.P.: data curation and visualization; A.P. and M.Z.: writing—original draft; V.C. and A.B.: conceptualization; writing—reviewing and editing; and supervision; All authors have read and approved the article. The authors declare that all data were generated in-house and that no paper mill was used.
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Lykhova, O., Zavelevich, M., Philchenkov, A. et al. Does insulin make breast cancer cells resistant to doxorubicin toxicity?. Naunyn-Schmiedeberg's Arch Pharmacol 396, 3111–3122 (2023). https://doi.org/10.1007/s00210-023-02516-3
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DOI: https://doi.org/10.1007/s00210-023-02516-3