Resveratrol inhibits cancer cell proliferation by impairing oxidative phosphorylation and inducing oxidative stress
Graphical abstract
Introduction
The beneficial role of several phytochemicals for multiple illnesses (cardiovascular events, obesity, diabetes and cancer) has been widely documented (Szkudelska and Szkudelski, 2010; Jiang et al., 2017). Particularly, resveratrol (trans-3,5,4′-tryhydroxystilbene, RSV) a natural polyphenol found in large quantities in grapes, berries and peanuts has shown multiple positive effects on normal cells (Mukherjee et al., 2010; Baarine et al., 2011). At 0.1–50 μM doses, RSV (a) activates signaling and transcription factors involved in cell-cycle regulation, apoptosis, angiogenesis, antioxidant mechanism; and down-regulates cyclooxygenase 2 and NF-kB in immune cells (Gao et al., 2001; Švajger and Jeras, 2012); (b) shows ROS scavenger capacity (increasing NOS expression and SIRT1 activation) improving mitochondrial function in heart (Turan et al., 2012) whereas in neurons, RSV shows a protective role against Alzheimer's and Parkinson's diseases in aging models (Richard et al., 2011); and (c) inhibits initiation, promotion and progression of cancer development with apparent low toxicity for normal cells (Kueck et al., 2007; Gwak et al., 2015).
In cancer cells, RSV (15–50 μM for 24–72 h) arrests the cellular cycle, induces apoptosis and promotes a massive reactive oxygen species (ROS) production (García-Zepeda et al., 2013). RSV also decreases the mRNA content (30–85%) of several glycolytic transporters (GLUT1) and enzymes (HKII, PFK-1 and PGM) and glycolytic flux (20–60%) in human hepatocellular carcinomas (HCC-LM3 and Bel-7402), diffuse large B-cell lymphoma (OCI-Ly1 and OCI-Ly18), breast cancer (MCF-7) and brain carcinomas (H460, HCC827 and H165) (Faber et al., 2006; Gomez et al., 2013; Jung et al., 2013; Dai et al., 2015). Although the mechanisms associated with the effect of RSV on glycolysis have not been clearly elucidated, it has been demonstrated that the activation of PI-3K/Akt/mTOR signaling pathway decreases the activities of the glucose transporter and several glycolytic enzymes in cancer cells (Faber et al., 2006; Kueck et al., 2007; Iqbal and Bamezai, 2012; Gwak et al., 2015). In addition, the inhibition of the PI-3K pathway by RSV also suppresses the accumulation of the key transcriptional regulator HIF-1α and of its glycolytic targets, leading to a lower glycolytic flux (Jung et al., 2013).
On the other hand, RSV (10–50 μM) increases the total oxygen consumption (30–80%) after 24–48 h as well as the contents of respiratory chain and mitochondrial biogenesis proteins in colon cancer cells (SW620, CaCo2, HCT116, HCEC, ICT, RPA) and mouse Lewis lung carcinoma (Blanquer-Rosselló et al., 2017; Saunier et al., 2017). As the mitochondrial membrane potential only increased by 10%, the data suggested that the RSV-induced oxygen uptake increase was not coupled to ATP synthesis. However, since the oxidative phosphorylation (OxPhos) flux was not assessed in these cancer cell lines, it is difficult to ascertain whether there are in fact RSV effects on mitochondrial function. This apparent mitochondrial function improvement at low RSV doses (20 μM, 24 h) has not been observed in non-cancer (epithelial) cells (Sheu et al., 2013). In addition, the effect of RSV on OxPhos, at doses in which cancer cell death occurs, has not been systematically assessed.
Therefore, the present study analyzed whether RSV only affects glycolysis or whether it also affects OxPhos flux, the predominant energy supplier in cancer cells under normoxia (Rodríguez-Enríquez et al., 2010; Hernández-Reséndiz et al., 2015; Pacheco-Velázquez et al., 2018). To this end, an integral analysis of the glycolytic and OxPhos enzymes contents, activities and fluxes was carried out in HeLa cells. ROS levels and antioxidant metabolites and enzymes as well as mitophagy processes were also examined. Our results clearly indicated that RSV, at doses near the growth IC50 values, acts as pro-oxidant drug inhibiting both OxPhos and glycolysis leading to ROS accumulation which in turn activated cellular death through mitophagy.
Section snippets
Chemicals
GR and LDH were purchased from Roche (Mannheim, Germany). Acetyl-CoA, ADP, ascorbate, antimycin, CHP, CCCP, DNCB, DTNB, 2-deoxyglucose, GSH, GSSG, glucose, glutamate, horse heart cytochrome, malate, NADH, NADP+, NADPH, NH4Cl, NBT, 2OG, oxaloacetate, oligomycin, rotenone, rhodamine 6G, t-BHP, trifluoroacetic acid, TMPD, triton X-100, xanthine and xanthine oxidase were from Sigma Chemical (St. Louis, MO, USA).
RSV solubility
Low RSV (Sigma Chemical, St. Louis, MO, USA) solubility in water has been documented
RSV effect on cancer cell growth
Exposure of cancer cells in bi-dimensional culture to 100–750 μM RSV for 48 h significantly decreased cell growth (Fig. 1A). Although a large amount of dead floating cells was observed, viability of attached cells was still high (> 80%); lower RSV concentrations were innocuous. Metastatic cancer cells showed slightly greater susceptibility to RSV than low-metastatic cancer cells (Table 1). Shorter exposure to RSV (12 or 24 h) at similar doses did not affect cancer cell proliferation and
Discussion
Anti-cancer effects of RSV (50–200 μM/4–48 h) have been documented (Kueck et al., 2007; Gwak et al., 2015). Its effects seem to be associated to its ability for (1) inducing apoptosis (Benitez et al., 2007); (2) inhibiting cell cycle (Benitez et al., 2007); and (3) promoting severe oxidative stress (Ji et al., 2018). At the metabolic level, RSV may affect both energy pathways, glycolysis (Faber et al., 2006; Gomez et al., 2013; Jung et al., 2013; Dai et al., 2015) as well as OxPhos, as
Conclusion
Our results suggested that the molecular mechanism by which RSV induces a growth arrest of HeLa cells is related principally to its pro-oxidant effect, which in turn induces ROS overload that surpasses the antioxidant machinery capacity and brings about mitochondrial function impairment. As treatment of cancer cells with antioxidants drugs has yielded negative outcomes and has been found to rather favor cancer progression (Tong et al., 2015; Assi, 2017), it seems that the use of pro-oxidant
Competing interests
The authors declare that they have no competing interests.
Funding statement
The present work was partially supported by grants from CONACyT-México to SRE (No. 283144) and RMS (239 930, 281 428).
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2024, Cancer LettersResveratrol inhibits HeLa cell proliferation by regulating mitochondrial function
2022, Ecotoxicology and Environmental SafetyCitation Excerpt :Thus, multiple studies focus on cellular level studies to explore the underlying mechanisms, and proliferation inhibition effects of RES on HeLa and other tumor cells are elaborated (Chang et al., 2015; Ho et al., 2020; Repossi et al., 2020). The related therapeutic mechanisms involve oxidative stress (Rodriguez-Enriquez et al., 2019), autophagy (Luyten et al., 2017), cellular pathway regulation (Liu et al., 2020), mitochondrial pathophysiology (Singh et al., 2021), DNA binding (Platella et al., 2020), and energy metabolism (Huang et al., 2019). Recently, Shaito et al. (2020) propose that RES has a "low-concentration antioxidant, high-concentration pro-oxidant" effect, emphasizing the importance of dose selection.
Metformin inhibits the tumor-promoting effect of low-dose resveratrol, and enhances the anti-tumor activity of high-dose resveratrol by increasing its reducibility in triple negative breast cancer
2022, Free Radical Biology and MedicineCitation Excerpt :Currently, there are two opposing viewpoints on the anti-tumor mechanism of resveratrol. One viewpoint is that resveratrol can promote the production of reactive oxygen species and induce apoptosis through pro-oxidation effects [27]. Another idea is that resveratrol inhibits tumor cell growth through its anti-oxidant effects [18,19].