Journal: Journal of the Egyptian National Cancer Institute

Article Title: Exploring potential additive effects of 5-fluorouracil, thymoquinone, and coenzyme Q10 triple therapy on colon cancer cells in relation to glycolysis and redox status modulation.

doi: 10.1186/s43046-025-00261-7

Figure Lengend Snippet: Fig. 8 Schematic representation of the proposed mechanism of action for the 5-FU, TQ and CQ10 triple therapy in colon cancer cells. The triple therapy demonstrates synergistic anticancer effects by modulating multiple pathways. (1) TQ enhances 5-FU chemosensitivity and (2) both drugs induce apoptosis by upregulating tumour suppressors (p21, p27, BAX, Cytochrome C, Caspase-3) and inhibiting oncogenes (CCND1, CCND3, BCL2, Survivin, PI3K/AKT/mTOR). Moreover, (3) both drugs promote aerobic phosphorylation and reduce chemoresistance by inhibiting LDHA and PDHK1, while increasing PDH activity, (4) thereby elevating oxidative stress and ROS-induced cell death. CQ10, despite its antioxidant properties, acts as a pro-oxidant in this context, as (5) increased ROS levels generated by 5-FU and TQ overwhelm CQ10 antioxidant capacity, (6) leading to the generation of additional free radicals by electron leakage. Moreover, CQ10 further contributes to the anticancer effects by (7) promoting HIF1α degradation, (8) attenuating the PI3K/AKT/mTOR pathway, inhibiting hypoxia, and enhancing ROS-induced apoptosis. (Arrows: Indicate activation or increased expression; T-bars: Indicate inhibition; 5-FU: 5-Fluorouracil; AKT: protein kinase B; BAX: BCL2-associated X protein; BCL2: B-cell lymphoma 2; CCND1: cyclin D1; CCND3: cyclin D3; CQ10: coenzyme Q10; HIF1α: hypoxia-inducible factor-1α; LDHA: lactate dehydrogenase A; mTOR; mammalian target of rapamycin; p21: Cyclin-dependent kinase inhibitor 1A; p27: Cyclin-dependent kinase inhibitor 1B; PDH: pyruvate dehydrogenase; PDHK1: pyruvate dehydrogenase kinase 1; ROS, reactive oxygen species; PI3K: phosphatidylinositol-3-kinase; and TQ: thymoquinone; Figure was created by BioRender.com)

Article Snippet: While GAPDH, CCND3, Cytochrome C, and PI3K-p85α were detected by mouse monoclonal antibodies, rabbit monoclonal antibodies were used for CCND1, p21, p27, BCL2, BAX, cleaved Caspase-3, PTEN, AKT1, mTOR, LDHA, PDH, PDHK1, and HIF1α (Cell Signaling Technology Inc.; MA, USA).

Techniques: Phospho-proteomics, Activity Assay, Generated, Activation Assay, Expressing, Inhibition

Journal: International Journal of Molecular Sciences

Article Title: All Three AKT Isoforms Can Upregulate Oxygen Metabolism and Lactate Production in Human Hepatocellular Carcinoma Cell Lines

doi: 10.3390/ijms25042168

Figure Lengend Snippet: The impact of single AKT isoforms on the inhibitory phosphorylation of PDH at Ser232 in HCC cells. Expression of AKT isoforms, levels of pyruvate dehydrogenase (PDH) phosphorylated at the inhibitory residue Ser232 (p-PDH) as well as total PDH expression were analyzed by Western blot ( A ) after the expression of constitutively activated AKT isoforms in Huh7 cells and ( B ) following the knockdown of individual AKT isoforms in Hep3B cells. The experiments were repeated three times and the representative results are shown.

Article Snippet: Thereafter, the proteins were blotted onto a nitrocellulose membrane and incubated with specific primary antibodies against AKT1 (#2938S, Cell Signaling Technology Inc., Danvers, MA, USA), AKT2 (#5239S, Cell Signaling Technology Inc., Danvers, MA, USA), AKT3 (#8018S, Cell Signaling Technology Inc., Danvers, MA, USA), PDH (#2784S, Cell Signaling Technology Inc., Danvers, MA, USA), p-PDH Ser232 (#15289S, Cell Signaling Technology Inc., Danvers, MA, USA) and GAPDH (#G0622, Santa Cruz Biotechnology Inc., Dallas, TX, USA).

Techniques: Phospho-proteomics, Expressing, Residue, Western Blot, Knockdown

Journal: International Journal of Molecular Sciences

Article Title: All Three AKT Isoforms Can Upregulate Oxygen Metabolism and Lactate Production in Human Hepatocellular Carcinoma Cell Lines

doi: 10.3390/ijms25042168

Figure Lengend Snippet: Model of the regulation of oxygen consumption and lactate production by the three AKT isoforms. AKT1, AKT2 and AKT3 increase oxygen consumption in HCC cells by a molecular mechanism leading to dephosphorylation of PDH at the inhibitory regulatory Ser232. The upregulation (blue arrows) of metabolic changes compared with control cells are shown.

Article Snippet: Thereafter, the proteins were blotted onto a nitrocellulose membrane and incubated with specific primary antibodies against AKT1 (#2938S, Cell Signaling Technology Inc., Danvers, MA, USA), AKT2 (#5239S, Cell Signaling Technology Inc., Danvers, MA, USA), AKT3 (#8018S, Cell Signaling Technology Inc., Danvers, MA, USA), PDH (#2784S, Cell Signaling Technology Inc., Danvers, MA, USA), p-PDH Ser232 (#15289S, Cell Signaling Technology Inc., Danvers, MA, USA) and GAPDH (#G0622, Santa Cruz Biotechnology Inc., Dallas, TX, USA).

Techniques: De-Phosphorylation Assay, Control