2. Academic Validation
  3. Metformin and thymoquinone co-treatment enhance 5-fluorouracil cytotoxicity by suppressing the PI3K/mTOR/HIF1α pathway and increasing oxidative stress in colon cancer cells

Metformin and thymoquinone co-treatment enhance 5-fluorouracil cytotoxicity by suppressing the PI3K/mTOR/HIF1α pathway and increasing oxidative stress in colon cancer cells

  • Biofactors. 2023 Mar 16. doi: 10.1002/biof.1947.
Wesam F Farrash 1,  Akhmed Aslam 1,  Riyad Almaimani 2,  Faisal Minshawi 1,  Hussain Almasmoum 1,  Aiman Alsaegh 1,  Mohammad S Iqbal 1,  Aisha Tabassum 1,  Mohamed E Elzubier 2,  Mahmoud Z El-Readi 2 3,  Amani A Mahbub 1,  Shakir Idris 1,  Bassem Refaat 1
Affiliations

Affiliations

  • 1 Laboratory Medicine Department, Faculty of Applied Medical Sciences, Umm Al-Qura University, Makkah, Saudi Arabia.
  • 2 Department of Biochemistry, Faculty of Medicine, Umm Al-Qura University, Makkah, Saudi Arabia.
  • 3 Biochemistry Department, Faculty of Pharmacy, Al-Azhar University, Assuit, Egypt.
PMID: 36929658 DOI: 10.1002/biof.1947
Abstract

This study investigated the chemotherapeutic effects of 5-fluorouracil (5-FU), metformin (Met), and/or thymoquinone (TQ) single/dual/triple therapies in the HT29, SW480 and SW620 colon Cancer (CRC) cell lines. Cell cycle/Apoptosis were measured by flow cytometry. The gene and protein expression of Apoptosis (PCNA/Survivin/Bax/Cytochrome-C/Caspase-3) and cell cycle (CCND1/CCND3/p21/p27) molecules, the PI3K/mTOR/HIF1α oncogenic pathway, and glycolysis regulatory enzymes were measured by quantitative-PCR and Western blot. Markers of oxidative stress were also measured by colorimetric assays. Although all treatments induced anti-cancer effects related to cell cycle arrest and Apoptosis, the triple therapy showed the highest pro-apoptotic actions that coincided with the lowest expression of CCND1/CCND3/PCNA/Survivin and the maximal increases in p21/p27/Bax/Cytochrome-C/Caspase-3 in all cell lines. The triple therapy also revealed the best suppression of the PI3K/mTOR/HIF1α pathway by increasing its endogenous inhibitors (PTEN/AMPKα) in all cell lines. Moreover, the lowest expression of Lactate Dehydrogenase and pyruvate dehydrogenase kinase-1 with the highest expression of pyruvate dehydrogenase were seen with the triple therapy, which also showed the highest increases in oxidative stress markers (ROS/RNS/MDA/protein carbonyl groups) alongside the lowest antioxidant levels (GSH/CAT) in all cell lines. In conclusion, this is the first study to reveal enhanced anti-cancer effects for metformin/thymoquinone in CRC that were superior to all monotherapies and the other dual therapies. However, the triple therapy approach showed the best tumoricidal actions related to cell cycle arrest and Apoptosis in all cell lines, possibly by enhancing oxidative glycolysis and augmenting oxidative stress through stronger modulation of the PI3K/mTOR/HIF1α oncogenic network.

Keywords

apoptosis; cell cycle; chemoresistance; glycolysis; oxidative stress.

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