Thioredoxin Reductase as a Potential Molecular Target for Anticancer Agents That Induce Oxidative Stress

Background: Thioredoxin reductase (TrxR) plays vital role in regulating cellular redox balance as well as redox-mediated signal transduction. Accumulating evidence supports that overactivation of TrxR is closely related to tumorigenesis and that targeting TrxR ablation reverses the growth of numerous malignant tumors, making TrxR a promising target for cancer chemotherapy. Thus, the discovery and development of molecules as promising anticancer agents that target TrxR is of great significance. Oridonin was shown to inhibit TrxR activity, but the detailed cellular mechanism is largely unknown. Objective: The study investigated the mechanism of action and underlying inhibitory properties of oridonin on TrxR in HeLa cells. Methods: A covalent docking was performed to reveal the possible interaction between oridonin and TrxR by Schrödinger Software Suite. TrxR activity was determined by 5,5’-dithiobis-2-nitrobenzoic acid reduction assay and endpoint insulin reduction assay. Sulforhodamine B and colony formation assay were employed to assess the viability and growth of cells. Reactive oxygen species level was measured by probe 2’, 7’-dichlorfluorescein diacetate, and dihydroethidium. Hoechst 33342 staining, caspase 3 activation, and fluorescein-5-isothiocyanate-conjugated Annexin V and propidium iodide double staining were used to evaluate apoptosis. Results: Here, we reported the oridonin as a potent inhibitor of TrxR. Inhibition of TrxR results in a decrease of thiols content and total glutathione elevates reactive oxygen species levels, and finally promotes oxidative stress-mediated apoptosis of cancer cells. Conclusion : Targeting TrxR by oridonin discloses a novel molecular mechanism underlying the biological action of oridonin and sheds light on developing oridonin as a potential tumor therapeutic agent.

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Oxidative stress response in regulatory and conventional T cells: a comparison between patients with chronic coronary syndrome and healthy subjects

Journal of Translational Medicine ◽

10.1186/s12967-021-02906-2 ◽

2021 ◽

Vol 19 (1) ◽

Author(s):

Anna K. Lundberg ◽

Rosanna W. S. Chung ◽

Louise Zeijlon ◽

Gustav Fernström ◽

Lena Jonasson

Keyword(s):

Oxidative Stress ◽

T Cells ◽

Mononuclear Cells ◽

Ex Vivo ◽

Thioredoxin Reductase ◽

Healthy Controls ◽

Antioxidant Genes ◽

Consistent Finding ◽

Coronary Syndrome ◽

Thioredoxin Reductase 1

Abstract Background Inflammation and oxidative stress form a vicious circle in atherosclerosis. Oxidative stress can have detrimental effects on T cells. A unique subset of CD4+ T cells, known as regulatory T (Treg) cells, has been associated with atheroprotective effects. Reduced numbers of Treg cells is a consistent finding in patients with chronic coronary syndrome (CCS). However, it is unclear to what extent these cells are sensitive to oxidative stress. In this pilot study, we tested the hypothesis that oxidative stress might be a potential contributor to the Treg cell deficit in CCS patients. Methods Thirty patients with CCS and 24 healthy controls were included. Treg (CD4+CD25+CD127−) and conventional T (CD4+CD25−, Tconv) cells were isolated and treated with increasing doses of H2O2. Intracellular ROS levels and cell death were measured after 2 and 18 h, respectively. The expression of antioxidant genes was measured in freshly isolated Treg and Tconv cells. Also, total antioxidant capacity (TAC) was measured in fresh peripheral blood mononuclear cells, and oxidized (ox) LDL/LDL ratios were determined in plasma. Results At all doses of H2O2, Treg cells accumulated more ROS and exhibited higher rates of death than their Tconv counterparts, p < 0.0001. Treg cells also expressed higher levels of antioxidant genes, including thioredoxin and thioredoxin reductase-1 (p < 0.0001), though without any differences between CCS patients and controls. Tconv cells from CCS patients were, on the other hand, more sensitive to oxidative stress ex vivo and expressed more thioredoxin reductase-1 than Tconv cells from controls, p < 0.05. Also, TAC levels were lower in patients, 0.97 vs 1.53 UAE/100 µg, p = 0.001, while oxLDL/LDL ratios were higher, 29 vs 22, p = 0.006. Conclusion Treg cells isolated from either CCS patients or healthy controls were all highly sensitive to oxidative stress ex vivo. There were signs of oxidant-antioxidant imbalance in CCS patients and we thus assume that oxidative stress may play a role in the reduction of Treg cells in vivo.

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Metalloproteomics for molecular target identification of protein-binding anticancer metallodrugs

Metallomics ◽

10.1039/d0mt00196a ◽

2020 ◽

Vol 12 (11) ◽

pp. 1627-1636

Author(s):

Tasha R. Steel ◽

Christian G. Hartinger

Keyword(s):

Protein Binding ◽

Anticancer Agents ◽

Target Identification ◽

Molecular Target ◽

Protein Targets ◽

Anticancer Metallodrugs

The development of the metallodrug pull-down as a metalloproteomic technique has enabled the identification of the protein targets of metal-based anticancer agents.

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Recent Advances in the Development of Thioredoxin Reductase Inhibitors as Anticancer Agents

Current Drug Targets ◽

10.2174/138945012803530224 ◽

2012 ◽

Vol 13 (11) ◽

pp. 1432-1444 ◽

Cited By ~ 30

Author(s):

Yang Liu ◽

Yijing Li ◽

Shenghui Yu ◽

Guisen Zhao

Keyword(s):

Anticancer Agents ◽

Thioredoxin Reductase ◽

Reductase Inhibitors ◽

Recent Advances

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Unifying mechanism for anticancer agents involving electron transfer and oxidative stress: Clinical implications

Medical Hypotheses ◽

10.1016/j.mehy.2006.08.046 ◽

2007 ◽

Vol 69 (3) ◽

pp. 510-516 ◽

Cited By ~ 40

Author(s):

Peter Kovacic

Keyword(s):

Oxidative Stress ◽

Electron Transfer ◽

Anticancer Agents ◽

Clinical Implications ◽

And Oxidative Stress

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Promotion of HeLa cells apoptosis by cynaropicrin involving inhibition of thioredoxin reductase and induction of oxidative stress

Free Radical Biology and Medicine ◽

10.1016/j.freeradbiomed.2019.03.014 ◽

2019 ◽

Vol 135 ◽

pp. 216-226 ◽

Cited By ~ 14

Author(s):

Tianyu Liu ◽

Junmin Zhang ◽

Xiao Han ◽

Jianqiang Xu ◽

Yueting Wu ◽

...

Keyword(s):

Oxidative Stress ◽

Hela Cells ◽

Thioredoxin Reductase

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Role of thioredoxin reductase 1 in dysplastic transformation of human breast epithelial cells triggered by chronic oxidative stress

Scientific Reports ◽

10.1038/srep36860 ◽

2016 ◽

Vol 6 (1) ◽

Cited By ~ 19

Author(s):

Chaoran Dong ◽

Lei Zhang ◽

Ruoxuan Sun ◽

Jianying Liu ◽

Hanwei Yin ◽

...

Keyword(s):

Oxidative Stress ◽

Epithelial Cells ◽

Thioredoxin Reductase ◽

Human Breast ◽

Breast Epithelial Cells ◽

Chronic Oxidative Stress ◽

Thioredoxin Reductase 1 ◽

Breast Epithelial ◽

Human Breast Epithelial Cells

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Oxidative Stress Response in Regulatory and Conventional T Cells: A Comparison Between Patients with Chronic Coronary Syndrome and Healthy Subjects

10.21203/rs.3.rs-110580/v1 ◽

2020 ◽

Author(s):

Anna K Lundberg ◽

Rosanna WS Chung ◽

Louise Zeijlon ◽

Gustav Fernström ◽

Lena Jonasson

Keyword(s):

Oxidative Stress ◽

T Cells ◽

Mononuclear Cells ◽

Ex Vivo ◽

Thioredoxin Reductase ◽

Healthy Controls ◽

Antioxidant Genes ◽

Consistent Finding ◽

Coronary Syndrome ◽

Thioredoxin Reductase 1

Abstract BackgroundInflammation and oxidative stress form a vicious circle in atherosclerosis. Oxidative stress can have detrimental effects on T cells. A unique subset of CD4+ T cells, known as regulatory T (Treg) cells, has been associated with atheroprotective effects. Reduced numbers of Treg cells is a consistent finding in patients with chronic coronary syndrome (CCS). However, it is unclear to what extent these cells are sensitive to oxidative stress. In the present study, we tested the hypothesis that oxidative stress might be a potential contributor to the Treg cell deficit in CCS patients. MethodsThirty patients with CCS and 24 healthy controls were included. Treg (CD4+CD25+CD127-) and conventional T (CD4+CD25-, Tconv) cells were isolated and treated with increasing doses of H2O2. Intracellular ROS levels and cell death were measured after 2 and 18 h, respectively. The expression of antioxidant genes was measured in freshly isolated Treg and Tconv cells. Alxo, total antioxidant capacity (TAC) was measured in fresh peripheral blood mononuclear cells. ResultsAt all doses of H2O2, Treg cells accumulated more ROS and exhibited higher rates of death than their Tconv counterparts, p < 0.0001. Treg cells also expressed higher levels of antioxidant genes, including thioredoxin and thioredoxin reductase-1 (p < 0.0001), though without any differences between CCS patients and controls. Tconv cells from CCS patients were, on the other hand, more sensitive to oxidative stress ex vivo and expressed more thioredoxin reductase-1 than Tconv cells from controls, p < 0.05. Also, TAC levels were lower in patients, 0.97 vs 1.53 UAE/100 µg, p = 0.001. ConclusionTreg cells isolated from either CCS patients or healthy controls were all highly sensitive to oxidative stress ex vivo. There were however signs of oxidant-antioxidant imbalance in CCS patients and we thus assume that oxidative stress plays a role in the reduction of Treg cells in vivo.

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Acetylated Thioredoxin Reductase 1 Resists Oxidative Inactivation

Frontiers in Chemistry ◽

10.3389/fchem.2021.747236 ◽

2021 ◽

Vol 9 ◽

Author(s):

David. E. Wright ◽

Nikolaus Panaseiko ◽

Patrick O’Donoghue

Keyword(s):

Oxidative Stress ◽

Thioredoxin Reductase ◽

Oxygen Species ◽

Site Specific ◽

Oxidative Inactivation ◽

Thioredoxin Reductase 1 ◽

Lysine Residues ◽

Canonical Amino Acid ◽

Genetic Code Expansion ◽

Low Activity

Thioredoxin Reductase 1 (TrxR1) is an enzyme that protects human cells against reactive oxygen species generated during oxidative stress or in response to chemotherapies. Acetylation of TrxR1 is associated with oxidative stress, but the function of TrxR1 acetylation in oxidizing conditions is unknown. Using genetic code expansion, we produced recombinant and site-specifically acetylated variants of TrxR1 that also contain the non-canonical amino acid, selenocysteine, which is essential for TrxR1 activity. We previously showed site-specific acetylation at three different lysine residues increases TrxR1 activity by reducing the levels of linked dimers and low activity TrxR1 tetramers. Here we use enzymological studies to show that acetylated TrxR1 is resistant to both oxidative inactivation and peroxide-induced multimer formation. To compare the effect of programmed acetylation at specific lysine residues to non-specific acetylation, we produced acetylated TrxR1 using aspirin as a model non-enzymatic acetyl donor. Mass spectrometry confirmed aspirin-induced acetylation at multiple lysine residues in TrxR1. In contrast to unmodified TrxR1, the non-specifically acetylated enzyme showed no loss of activity under increasing and strongly oxidating conditions. Our data suggest that both site-specific and general acetylation of TrxR1 regulate the enzyme’s ability to resist oxidative damage.

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Inhibition of Glutathione and Thioredoxin Metabolism Enhances Sensitivity to Perifosine in Head and Neck Cancer Cells

Journal of Oncology ◽

10.1155/2009/519563 ◽

2009 ◽

Vol 2009 ◽

pp. 1-10 ◽

Cited By ~ 25

Author(s):

Andrean L. Simons ◽

Arlene D. Parsons ◽

Katherine A. Foster ◽

Kevin P. Orcutt ◽

Melissa A. Fath ◽

...

Keyword(s):

Oxidative Stress ◽

Head And Neck Cancer ◽

Head And Neck ◽

Neck Cancer ◽

Reductase Activity ◽

Thioredoxin Reductase ◽

Buthionine Sulfoximine ◽

Human Head ◽

Akt Inhibitor ◽

Simultaneous Treatment

The hypothesis that the Akt inhibitor, perifosine (PER), combined with inhibitors of glutathione (GSH) and thioredoxin (Trx) metabolism will induce cytotoxicity via metabolic oxidative stress in human head and neck cancer (HNSCC) cells was tested. PER induced increases in glutathione disulfide (%GSSG) in FaDu, Cal-27, and SCC-25 HNSCCs as well as causing significant clonogenic cell killing in FaDu and Cal-27, which was suppressed by simultaneous treatment with N-acetylcysteine (NAC). An inhibitor of GSH synthesis, buthionine sulfoximine (BSO), sensitized Cal-27 and SCC-25 cells to PER-induced clonogenic killing as well as decreased total GSH and increased %GSSG. Additionally, inhibition of thioredoxin reductase activity (TrxRed) with auranofin (AUR) was able to induce PER sensitization in SCC-25 cells that were initially refractory to PER. These results support the conclusion that PER induces oxidative stress and clonogenic killing in HNSCC cells that is enhanced with inhibitors of GSH and Trx metabolism.

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