Objective:The objective of the current study was to evaluate the chemosensitizing capacity of auranofin(AF),a gold(I)complex traditionally used in rheumatoid arthritis treatment,in potentiating the cytotoxic effects o...Objective:The objective of the current study was to evaluate the chemosensitizing capacity of auranofin(AF),a gold(I)complex traditionally used in rheumatoid arthritis treatment,in potentiating the cytotoxic effects of doxorubicin(DOX)in melanoma cell models,specifically drug-sensitive(B16F10)and multidrug-resistant(B16F10/ADR)variants.Methods:Experimental measurements,including in vitro cytotoxicity and apoptosis assays,surface plasmon resonance(SPR),immunoblotting assays,as well as theoretical calculations,such as molecular docking and molecular dynamics(MD)simulations,were used to systematically delineate the interaction dynamics between AF and thioredoxin reductase 1(TrxR1).The anti-tumor efficacy of co-treatment with AF and DOX was assessed by examining cell viability and apoptotic rates.Results:Co-treatment with AF and DOX significantly increased anti-tumor efficacy,as evidenced by reduced cell viability and increased apoptotic rates.This synergistic effect was attributed to inhibition of TrxR1 by AF,which compromised tumor cell antioxidant defenses and elevated intracellular reactive oxygen species(ROS),thereby enhancing apoptotic pathways.Notably,AF treatment mitigated the heightened TrxR activity in DOX-resistant cells,intensifying the pro-oxidant effects of DOX,leading to increased ROS production and cell death.The data also showed that AF binds with high affinity to the selenocysteine residue within the catalytic site of TrxR1,which partially overlapped with the binding site of the endogenous substrate,thioredoxin(Trx),but with greater avidity.This unique binding configuration impedes the reduction of Trx by TrxR1,triggering an apoptotic response in cancer cells.Conclusions:This study underscores the chemosensitizing potential of AF in overcoming multidrug resistance in cancer therapy through redox modulation.The molecular mechanism of action underlying AF on TrxR1 demonstrated the unique binding configuration that impedes the reduction of Trx by TrxR1 and instigates an apoptotic response in cancer cells.These findings pave the way for the clinical application of AF as a chemosensitizer,offering a novel approach to augment the efficacy of existing chemotherapy regimens.展开更多
Objective:To investigate the synergistic effects of auranofin and schisandrin A(SA)on cell proliferation inhibition and apoptosis induction in human hepatocellular carcinoma Hep3B cells.Methods:Cell viability was asse...Objective:To investigate the synergistic effects of auranofin and schisandrin A(SA)on cell proliferation inhibition and apoptosis induction in human hepatocellular carcinoma Hep3B cells.Methods:Cell viability was assessed using MTT to determine the synergistic effects of auranofin and SA.Three-dimensional(3D)culture models were used to evaluate the effects on spheroid structure and size.Apoptosis was analyzed by flow cytometry for sub-G1 populations,annexin V staining,and Western blotting for apoptotic markers.Reactive oxygen species(ROS)production was measured using DCF-DA staining.Results:Our results showed that combined treatment with auranofin and SA led to a significant reduction in cell viability compared with either compound alone,with isobologram analysis confirming their synergistic interactions.Under 3D culture conditions,auranofin and SA disrupted the compact structure of spheroids,leading to a loosened and disorganized morphology at the periphery,which appeared as an increase in spheroid size.Moreover,the induction of apoptosis by auranofin and SA was evidenced by elevated sub-G1 phase populations,increased annexin V-positive cells,and upregulation of apoptotic markers such as cleaved poly(ADPribose)polymerase 1 and cleaved caspase-3.Notably,auranofin combined with SA markedly enhanced ROS production,which was mitigated by the ROS scavenger N-acetylcysteine.Additionally,the phosphoinositide 3-kinase(PI3K)/Akt signaling pathway was downregulated in response to auranofin and SA treatment,and further apoptotic effects were observed following PI3K inhibition with LY294002.Conclusions:Auranofin combined with SA promotes apoptosis of hepatocellular carcinoma via ROS generation and inhibition of the PI3K/Akt pathway.展开更多
Neurodegenerative disorders including Alzheimer＇s disease are characterized by chronic in- flammation in the central nervous system. The two main glial types involved in inflammatory reactions are microglia and astro...Neurodegenerative disorders including Alzheimer＇s disease are characterized by chronic in- flammation in the central nervous system. The two main glial types involved in inflammatory reactions are microglia and astrocytes. While these cells normally protect neurons by providing nutrients and growth factors, disease specific stimuli can induce glial secretion of neurotoxins. It has been hypothesized that reducing glia-mediated inflammation could diminish neuronal loss. This hypothesis is supported by observations that chronic use of non-steroidal anti-inflamma- tory drugs （NSAIDs） is linked with lower incidences of neurodegenerative disease. It is possible that the NSAIDs are not potent enough to appreciably reduce chronic neuroinflammation after disease processes are fully established. Gold thiol compounds, including auranofin, comprise an- other class of medications effective at reducing peripheral inflammation. We have demonstrated that auranofin inhibits human microglia- and astrocyte-mediated neurotoxicity. Other drugs which are currently used to treat peripheral inflammatory conditions could be helpful in neu- rodegenerative disease. Three different classes of anti-inflammatory compounds, which have a potential to inhibit neuroinflammation are highlighted below.展开更多
Today colorectal cancer(CRC)is one of the leading causes of cancer death worldwide.This disease is poorly chemo-sensitive toward the existing medical treatments so that new and more effective therapeutic agents are ur...Today colorectal cancer(CRC)is one of the leading causes of cancer death worldwide.This disease is poorly chemo-sensitive toward the existing medical treatments so that new and more effective therapeutic agents are urgently needed and intensely sought.Platinum drugs,oxaliplatin in particular,were reported to produce some significant benefit in CRC treatment,triggering the general interest of medicinal chemists and oncologists for metal-based compounds as candidate anti-CRC drugs.Within this frame,gold compounds and,specifically,the established antiarthritic drug auranofin with its analogs,form a novel group of promising anticancer agents.Owing to its innovative mechanism of action and its favorable pharmacological profile,auranofin together with its derivatives are proposed here as novel experimental agents for CRC treatment,capable of overcoming resistance to platinum drugs.Some encouraging results in this direction have already been obtained.A few recent studies demonstrate that the action of auranofin may be further potentiated through the preparation of suitable pharmaceutical formulations capable of protecting the gold pharmacophore from unselective reactivity or through the design of highly synergic drug combinations.The perspectives of the research in this field are outlined.展开更多
AIM To study the sensitivity and antioxidant enzyme response in two clinical isolates of Entamoeba histolytica(E. histolytica) during treatment with antiamoebic drugs,auranofin and metronidazole. METHODS E. histolytic...AIM To study the sensitivity and antioxidant enzyme response in two clinical isolates of Entamoeba histolytica(E. histolytica) during treatment with antiamoebic drugs,auranofin and metronidazole. METHODS E. histolytica were isolated from stool samples and maintained in Robinson's biphasic culture medium. Clinial isolates were maintained in xenic culture medium,and harvested for determination of minimum inhibitory concentrations to the two antiamoebic drugs,Metronidazole and Auranofin using microtiter plate tests. The percent survival of the two isolates were determined using the trypan blue cell count. Isolate 980 was treated with 70 μmol/L and 2 μmol/L while isolate 989 was treated with 20 μmol/L and 0.5 μmol/L of metronidazole and auranofin respectively for 24 h. Fifty thousand cells of each isolate were harvested after 24 h of treatment for analysis of the mRNA expressions of the antioxidant enzymes,thioredoxin reductase,peroxiredoxin and FeSOD using the specific primers. Cell lysate was used for determination of enzyme activity of thioredoxin reductase by measuring DTNB reduction spectrophotometrically at412 nm.RESULTS Minimum inhibitory concentration of the clinical isolates 980 and 989 for auranofin was 3 μmol/L and 1 μmol/L respectively while that for metronidazole was 80 μmol/L and 30 μmol/L respectively. Thioredoxin reductase,peroxiredoxin and FeSOD expression levels were significantly reduced in the isolate 980 when treated with Auranofin. Metronidazole treatment showed a down regulation of thioredoxin reductase. Though not significant both at the mRNA and the enzyme activity levels. Peroxiredoxin and FeSOD however remained unchanged. Auranofin treatment of isolate 989,showed an upregulation in expression of thioredoxin reductase while Peroxiredoxin and FeSOD did not show any change in expression. Upon treatment with metronidazole,isolate 989 showed an increase in thioredoxin reductase expression. Peroxiredoxin and FeSOD expressions however remain unchanged both at mRNA and enzyme activity level.CONCLUSION Clinical isolates from New Delhi NCR region show different sensitivities to antiamoebic drugs. Auranofin is effective against isolate showing higher tolerance to metronidazole as shown by its inhibition in thioredoxin reductase activity.展开更多
基金supported by the National Natural Science Foundation of China(Grant Nos.82261138630,32171390,and 32201154)the Natural Science Foundation of Guangdong Province(Grant Nos.2023B1515020104 and 2024A1515011244).
文摘Objective:The objective of the current study was to evaluate the chemosensitizing capacity of auranofin(AF),a gold(I)complex traditionally used in rheumatoid arthritis treatment,in potentiating the cytotoxic effects of doxorubicin(DOX)in melanoma cell models,specifically drug-sensitive(B16F10)and multidrug-resistant(B16F10/ADR)variants.Methods:Experimental measurements,including in vitro cytotoxicity and apoptosis assays,surface plasmon resonance(SPR),immunoblotting assays,as well as theoretical calculations,such as molecular docking and molecular dynamics(MD)simulations,were used to systematically delineate the interaction dynamics between AF and thioredoxin reductase 1(TrxR1).The anti-tumor efficacy of co-treatment with AF and DOX was assessed by examining cell viability and apoptotic rates.Results:Co-treatment with AF and DOX significantly increased anti-tumor efficacy,as evidenced by reduced cell viability and increased apoptotic rates.This synergistic effect was attributed to inhibition of TrxR1 by AF,which compromised tumor cell antioxidant defenses and elevated intracellular reactive oxygen species(ROS),thereby enhancing apoptotic pathways.Notably,AF treatment mitigated the heightened TrxR activity in DOX-resistant cells,intensifying the pro-oxidant effects of DOX,leading to increased ROS production and cell death.The data also showed that AF binds with high affinity to the selenocysteine residue within the catalytic site of TrxR1,which partially overlapped with the binding site of the endogenous substrate,thioredoxin(Trx),but with greater avidity.This unique binding configuration impedes the reduction of Trx by TrxR1,triggering an apoptotic response in cancer cells.Conclusions:This study underscores the chemosensitizing potential of AF in overcoming multidrug resistance in cancer therapy through redox modulation.The molecular mechanism of action underlying AF on TrxR1 demonstrated the unique binding configuration that impedes the reduction of Trx by TrxR1 and instigates an apoptotic response in cancer cells.These findings pave the way for the clinical application of AF as a chemosensitizer,offering a novel approach to augment the efficacy of existing chemotherapy regimens.
基金supported by the National Research Foundation of Korea grant funded by the Korea government(RS-2023-00213236).
文摘Objective:To investigate the synergistic effects of auranofin and schisandrin A(SA)on cell proliferation inhibition and apoptosis induction in human hepatocellular carcinoma Hep3B cells.Methods:Cell viability was assessed using MTT to determine the synergistic effects of auranofin and SA.Three-dimensional(3D)culture models were used to evaluate the effects on spheroid structure and size.Apoptosis was analyzed by flow cytometry for sub-G1 populations,annexin V staining,and Western blotting for apoptotic markers.Reactive oxygen species(ROS)production was measured using DCF-DA staining.Results:Our results showed that combined treatment with auranofin and SA led to a significant reduction in cell viability compared with either compound alone,with isobologram analysis confirming their synergistic interactions.Under 3D culture conditions,auranofin and SA disrupted the compact structure of spheroids,leading to a loosened and disorganized morphology at the periphery,which appeared as an increase in spheroid size.Moreover,the induction of apoptosis by auranofin and SA was evidenced by elevated sub-G1 phase populations,increased annexin V-positive cells,and upregulation of apoptotic markers such as cleaved poly(ADPribose)polymerase 1 and cleaved caspase-3.Notably,auranofin combined with SA markedly enhanced ROS production,which was mitigated by the ROS scavenger N-acetylcysteine.Additionally,the phosphoinositide 3-kinase(PI3K)/Akt signaling pathway was downregulated in response to auranofin and SA treatment,and further apoptotic effects were observed following PI3K inhibition with LY294002.Conclusions:Auranofin combined with SA promotes apoptosis of hepatocellular carcinoma via ROS generation and inhibition of the PI3K/Akt pathway.
基金supported by a grant from the Jack BrownFamily Alzheimer’s Disease Research Foundation
文摘Neurodegenerative disorders including Alzheimer＇s disease are characterized by chronic in- flammation in the central nervous system. The two main glial types involved in inflammatory reactions are microglia and astrocytes. While these cells normally protect neurons by providing nutrients and growth factors, disease specific stimuli can induce glial secretion of neurotoxins. It has been hypothesized that reducing glia-mediated inflammation could diminish neuronal loss. This hypothesis is supported by observations that chronic use of non-steroidal anti-inflamma- tory drugs （NSAIDs） is linked with lower incidences of neurodegenerative disease. It is possible that the NSAIDs are not potent enough to appreciably reduce chronic neuroinflammation after disease processes are fully established. Gold thiol compounds, including auranofin, comprise an- other class of medications effective at reducing peripheral inflammation. We have demonstrated that auranofin inhibits human microglia- and astrocyte-mediated neurotoxicity. Other drugs which are currently used to treat peripheral inflammatory conditions could be helpful in neu- rodegenerative disease. Three different classes of anti-inflammatory compounds, which have a potential to inhibit neuroinflammation are highlighted below.
基金funding the project“Advanced mass spectrometry tools for cancer research:novel applications in proteomics,metabolomics and nanomedicine”(Multi-user Equipment Program 2016,Ref.code 19650)the Beneficentia Stiftung,Vaduz(BEN2019/48 and University of Pisa(Rating Ateneo 2019-2020)for the financial support+1 种基金supported by the University of Pisa under the“PRA-Progetti di Ricerca di Ateneo”Institutional Research Grants-Project no.PRA_2020_58“Agenti innovativi e nanosistemi per target molecolari nell’ambito dell’oncologia di precisione”to Marzo Tthe financial support(two-year fellowship for Italy“Marcello e Rosina Soru”-Project Code:23852).
文摘Today colorectal cancer(CRC)is one of the leading causes of cancer death worldwide.This disease is poorly chemo-sensitive toward the existing medical treatments so that new and more effective therapeutic agents are urgently needed and intensely sought.Platinum drugs,oxaliplatin in particular,were reported to produce some significant benefit in CRC treatment,triggering the general interest of medicinal chemists and oncologists for metal-based compounds as candidate anti-CRC drugs.Within this frame,gold compounds and,specifically,the established antiarthritic drug auranofin with its analogs,form a novel group of promising anticancer agents.Owing to its innovative mechanism of action and its favorable pharmacological profile,auranofin together with its derivatives are proposed here as novel experimental agents for CRC treatment,capable of overcoming resistance to platinum drugs.Some encouraging results in this direction have already been obtained.A few recent studies demonstrate that the action of auranofin may be further potentiated through the preparation of suitable pharmaceutical formulations capable of protecting the gold pharmacophore from unselective reactivity or through the design of highly synergic drug combinations.The perspectives of the research in this field are outlined.
基金Supported by WOS-A grant from Department of Science and Technology,New Delhi,Government of India to Iyer LR,No.SR/WOS-A/LS-98/2007"Programme support on molecular parasitology",Department of Biotechnology,New Delhi,India to Paul J,No.BT/01/CEIB/11/v/08
文摘AIM To study the sensitivity and antioxidant enzyme response in two clinical isolates of Entamoeba histolytica(E. histolytica) during treatment with antiamoebic drugs,auranofin and metronidazole. METHODS E. histolytica were isolated from stool samples and maintained in Robinson's biphasic culture medium. Clinial isolates were maintained in xenic culture medium,and harvested for determination of minimum inhibitory concentrations to the two antiamoebic drugs,Metronidazole and Auranofin using microtiter plate tests. The percent survival of the two isolates were determined using the trypan blue cell count. Isolate 980 was treated with 70 μmol/L and 2 μmol/L while isolate 989 was treated with 20 μmol/L and 0.5 μmol/L of metronidazole and auranofin respectively for 24 h. Fifty thousand cells of each isolate were harvested after 24 h of treatment for analysis of the mRNA expressions of the antioxidant enzymes,thioredoxin reductase,peroxiredoxin and FeSOD using the specific primers. Cell lysate was used for determination of enzyme activity of thioredoxin reductase by measuring DTNB reduction spectrophotometrically at412 nm.RESULTS Minimum inhibitory concentration of the clinical isolates 980 and 989 for auranofin was 3 μmol/L and 1 μmol/L respectively while that for metronidazole was 80 μmol/L and 30 μmol/L respectively. Thioredoxin reductase,peroxiredoxin and FeSOD expression levels were significantly reduced in the isolate 980 when treated with Auranofin. Metronidazole treatment showed a down regulation of thioredoxin reductase. Though not significant both at the mRNA and the enzyme activity levels. Peroxiredoxin and FeSOD however remained unchanged. Auranofin treatment of isolate 989,showed an upregulation in expression of thioredoxin reductase while Peroxiredoxin and FeSOD did not show any change in expression. Upon treatment with metronidazole,isolate 989 showed an increase in thioredoxin reductase expression. Peroxiredoxin and FeSOD expressions however remain unchanged both at mRNA and enzyme activity level.CONCLUSION Clinical isolates from New Delhi NCR region show different sensitivities to antiamoebic drugs. Auranofin is effective against isolate showing higher tolerance to metronidazole as shown by its inhibition in thioredoxin reductase activity.
