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Novel phloretin-based combinations targeting glucose metabolism in hepatocellular carcinoma through GLUT2/PEPCK axis of action: in silico molecular modelling and in vivo studies

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Abstract

Hepatocellular carcinoma (HCC) is commonly associated with disturbances in glucose metabolism and enhanced glycolysis. However, a controversial role for gluconeogenesis was reported to be tumor-promoting and tumor-suppressive. We investigated novel anti-HCC treatments through either the simultaneous inhibition of glycolysis and gluconeogenesis by “phloretin” and “sodium meta-arsenite”, respectively (Combination 1); or the concurrent inhibition of glycolysis and induction of gluconeogenesis by phloretin and dexamethasone, respectively, (combination 2). A total of 110 Swiss albino mice were divided into eleven groups, HCC was induced by N, N-dimethyl-4-aminoazobenzene. We have measured the expression of the glucose transporter 2 (GLUT2), Phosphoenolpyruvate carboxykinases (PEPCK), Caspase-3, Beclin 1, Cyclin D1, and cytokeratin 18 genes; blood glucose and ATP levels; alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities. Furthermore, in silico molecular docking was performed to investigate the potential drug-receptor interactions. Histologically, the phloretin-based combinations resulted in a significant regression of malignant tissue compared to various treatments. GLUT2 and PEPCK mRNA analysis indicated successful off/on modulation of glycolysis and gluconeogenesis. Docking confirmed the potent binding between phloretin, sodium meta-arsenite, and dexamethasone with GLUT2, PEPCK, and Retinoid X Receptor Alpha, respectively. Molecularly, Combination 2 resulted in the highest reduction in cyclin D1, cytokeratin 18, and Beclin 1 expression contemporaneously with the upregulation in Caspase-3 levels. Biochemically, both combinations caused a significant reduction in ATP levels, ALT, and AST activity compared to the other groups. In conclusion, we propose two novel phloretin-based combinations that can be used in treating HCC through the regulation of glucose metabolism and ATP production.

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Availability of supporting data

The data presented in this study are available on request from the corresponding author.

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Authors and Affiliations

  1. Department of Clinical Trial Research Unit and Drug Discovery, Egyptian Liver Research Institute and Hospital (ELRIAH), Mansoura, Egypt

    Alaa Elmetwalli

  2. Microbiology Division, Higher Technological Institute of Applied Health Sciences, Egyptian Liver Research Institute and Hospital (ELRIAH), Mansoura, Egypt

    Alaa Elmetwalli

  3. Laboratory Specialist, Alexandria, Egypt

    Neamat H. Kamosh & Rania El Safty

  4. Department of Applied Medical Chemistry, Medical Research Institute, Alexandria University, Alexandria, Egypt

    Amany I. Youssef & Tarek El-Sewedy

  5. Department of Histochemistry and Cell Biology, Medical Research Institute, Alexandria University, Alexandria, Egypt

    Mohammed M. Salama

  6. Experimental Animal Unit, Medical Research Institute, Alexandria University, Alexandria, Egypt

    Khaled M. Abd El-Razek

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Contributions

Conceptualization, N.H.K, A.I.Y., A.E., T.E.S.,; Formal analysis, M.M.S, K.M.A.E; Investigation, N.H.K, A.I.Y.,; Project administration, A.I.Y., A.E., T.E.S.,; Software, N.H.K, A.I.Y.; Validation A.I.Y., A.E., T.E.S.; Visualization, A.I.Y., A.E., T.E.S.; Writing—original draft, A.E.; Writing—review and editing, A.E, T.E.S; all authors have read and agreed to the published version of the manuscript. The authors declare that all data were generated in-house and that no paper mill was used.

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Correspondence to Alaa Elmetwalli.

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The study was conducted according to the guidelines of ARRIVE and approved by the Ethical Committee at the Medical Research Institute, Alexandria University.

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Elmetwalli, A., Kamosh, N.H., El Safty, R. et al. Novel phloretin-based combinations targeting glucose metabolism in hepatocellular carcinoma through GLUT2/PEPCK axis of action: in silico molecular modelling and in vivo studies. Med Oncol 41, 12 (2024). https://doi.org/10.1007/s12032-023-02236-x

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