A Novel and Specific NADPH Oxidase-1 (Nox1) Small-Molecule Inhibitor Blocks the Formation of Functional Invadopodia in Human Colon Cancer Cells

Gianni, Davide; Taulet, Nicolas; Zhang, Hui; DerMardirossian, Céline; Kister, Jeremy; Martinez, Luis; Roush, William; Brown, Steven J; Bokoch, Gary M.; Rosen, Hugh

doi:10.1021/cb100219n

Cited by 170 publications

(158 citation statements)

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“…APO is one of the most frequently used NOX inhibitors and may also act as an antioxidant (13). However, because of the lack of specificity of DPI and APO, novel and more selective NOX inhibitors, such as VAS, VAS3947, and 2-acetylphenothiazine (2APT), have been developed (9,35,40). Because these NOX inhibitors have been used in numerous physiological, cell biological, and biochemical studies, knowing the effects of these compounds on targets unrelated to NOX can be quite helpful and informative.…”

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confidence: 99%

The NADPH oxidase inhibitor diphenyleneiodonium activates the human TRPA1 nociceptor

Suzuki

Hatano

Muraki

et al. 2014

American Journal of Physiology-Cell Physiology

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Transient receptor potential ankyrin 1 (TRPA1) is a Ca 2ϩ -permeable nonselective cation channel expressed in neuronal and nonneuronal cells and plays an important role in acute and inflammatory pain. Here, we show that an NADPH oxidase (NOX) inhibitor, diphenyleneiodonium (DPI), functions as a TRPA1 activator in human embryonic kidney cells expressing human TRPA1 (HEK-TRPA1) and in human fibroblast-like synoviocytes. Application of DPI at 0.03-10 M induced a Ca 2ϩ response in HEK-TRPA1 cells in a concentration-dependent manner. The Ca 2ϩ response was effectively blocked by a selective TRPA1 antagonist, HC-030031 (HC). In contrast, DPI had no effect on HEK cells expressing TRPV1-V4 or TRPM8. Four other NOX inhibitors, apocynin (APO), VAS2870 (VAS), plumbagin, and 2-acetylphenothiazine, also induced a Ca 2ϩ response in HEK-TRPA1 cells, which was inhibited by pretreatment with HC. In the presence of 5 mM glutathione, the Ca 2ϩ response to DPI was effectively reduced. Moreover, mutation of cysteine 621 in TRPA1 substantially inhibited the DPIinduced Ca 2ϩ response, while it did not inhibit the APO-and VASinduced responses. The channel activity was induced by DPI in excised membrane patches with both outside-out and inside-out configurations. Internal application of neomycin significantly inhibited the DPI-induced inward currents. In inflammatory synoviocytes with TRPA1, DPI evoked a Ca 2ϩ response that was sensitive to HC. In mice, intraplantar injection of DPI caused a pain-related response which was inhibited by preadministration with HC. Taken together, our findings demonstrate that DPI and other NOX inhibitors activate human TRPA1 without mediating NOX. calcium channel; transient receptor potential; NADPH oxidase inhibitors TRANSIENT RECEPTOR POTENTIAL ANKYRIN 1 (TRPA1), a Ca 2ϩ -permeable nonselective cation channel widely expressed in neuronal and nonneuronal cells, is activated by noxious cold, mechanical stimulation, irritant chemicals, and some clinical drugs (1,7,12,17,26,28,30,36). Because transgenic mice lacking TRPA1 have suppressed sensitivity to mechanical stimulation, cold stimuli, and TNF␣-induced mechanical hyperalgesia (5,8,20,23,33), the channel has been proposed to be a nociceptor mediating acute and inflammatory pain (3,23,33). Mutagenesis studies have revealed that the cysteine residues 414, 421, and 621 of human TRPA1 are targeted by physiological and nonphysiological electrophilic compounds that activate the channel (14, 26). In contrast, nonelectrophilic compounds, such as ⌬9-tetrahydrocannabinol, nicotine, and menthol, activate TRPA1 via unknown mechanisms (17,18,38). Because numerous structurally unrelated compounds stimulate TRPA1, greater attention should be given to whether common clinical drugs and experimental pharmacological agents activate the channel.The NADPH oxidases (NOXs), a family of transmembrane proteins comprising seven members (NOX1-NOX5, DUOX1, and DUOX2), function as transmembrane electron transporters that use cytosolic NADPH as electron donor and oxygen as electron a...

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confidence: 99%

The NADPH oxidase inhibitor diphenyleneiodonium activates the human TRPA1 nociceptor

Suzuki

Hatano

Muraki

et al. 2014

American Journal of Physiology-Cell Physiology

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“…Expression of Nox1 seems to be relatively high in colon epithelial cells than that in other normal cell types (19). In colon cancer cells, the known function of Nox1 is conflicting: one study suggested that Nox1 was downstream of 12-HETE and was responsible for cell proliferation but not for cell spreading (20); another study suggested that Nox1 was responsible for cell invasion (6). In this study, we inves-…”

Section: Introductionmentioning

confidence: 91%

“…To answer this question, we performed immunohistochemistry using Nox1-specific antibody on tissue arrays, which includes normal and cancer tissues Grades 1 to 3, Stages I -IV, and various TNM stages (Table 1) Nox1 levels are higher in primary SW480 than that in metastatic SW620 cells Because ROS plays a key role in cell motility and invasion (21), inhibition of Nox1 was previously shown to inhibit invadopodia formation in colon cancer cells (6). We next examined Nox1 expression levels in SW480 and SW620 cells.…”

Section: Nox1 Level Is Not Associated With Colon Cancer Progressionmentioning

confidence: 99%

“…Recent studies have shown that Nox1 plays crucial roles in cancer development including transformation and invasion, (2,3) hence Nox1 was proposed to be a therapeutic target (4,5). A number of Nox1 inhibitors have been developed for various diseases (6)(7)(8), among which diphenyliodonium is an irreversible nonspecific inhibitor (9), VAS2870 is a Nox1 inhibitor identified via high-throughput screening (10), and apocynin is a natural compound that was widely used to inhibit Nox family activities (11,12).…”

Section: Introductionmentioning

confidence: 99%

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Association of Nox1 and Vinculin With Colon Cancer Progression

Sun

Liu

2013

Cancer Investigation

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Nox1 mRNA, protein, and activities were compared in the paired primary and metastatic colon adenocarcinoma cell lines SW480 and SW620, and in normal colon tissues and colon cancer tissues. Our results demonstrated that Nox1 levels were higher in the primary SW480 cells than that in metastatic SW620 cells and were not associated with colon cancer progression. We further discovered that vinculin protein level in SW620 was much higher than that in SW480 cells, whereas E-cadherin was lower. We conclude that vinculin and E-cadherin, but not Nox1, may serve as biomarkers for colon cancer progression.

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“…With regard to the available Nox inhibitors, there is much difference in opinion with regard to the most specific and efficient, and the choice is also limited by suitability to a particular cell type as well as the nature of the downstream effect of interest. Some compounds branded as Nox inhibitors, appear to function more preferentially as an anti-oxidant or free-radical scavenger (Heumuller et al, 2008), 2006), GKT136901 (Page P et al, 2008), ML171 (Gianni et al, 2010), ebselen and its analogs (Smith et al, 2012). It is possible that one or several of these, will be shown to demonstrate definitive, specific Nox inhibition.…”

Section: Quantitative Reverse Transcriptase Polymerase Chain Reactionmentioning

confidence: 99%

FLT3-driven redox-modulation of Ezrin regulates leukaemic cell migration

Corcoran

Cotter

2012

Free Radical Research

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Access to the full text of the published version may require a subscription. and ideas for this work, and who supported and believed in me throughout -I am grateful to have had the opportunity to learn from the best. I must also give a huge thank you to Kate Cotter, who made me feel at home from day one, and who has continued to be a tireless source of energy and assistance. RightsI was fortunate to interact with a vibrant, dynamic group in the Cotter Lab and I owe a huge thanks to every single member, past-Claire, Ruth, Carolyn, Chris, Ashley, To all the friends that I have made in U.C.C. and beyond during this process, as well as those friends lucky enough not to be involved but who still got to listen to all that whingeing-thank you! Lastly, I am most indebted to those closest to me. My mam -who is always ready to listen, comfort and reassure, and as a result probably knows more than she ever wanted to about cells and reactive oxygen species! My dad, a real scientist, who never stops teaching and never stops learning, and is still the person I turn to with my "homework" questions! I am so grateful to you both for your wisdom, encouragement and patience; truly I could not have achieved this without you. ToSinead and Dara, for all the much needed distractions and laughs, and for even being interested! And to John, for being there throughout, for the joy and the inspiration. To everyone still waiting for good news or the right medicine 'Happy is he who gets to know the reasons for things.' -Virgil DeclarationThis thesis has not been submitted in whole or in part to this or any other university for any degree and is, unless otherwise stated, the original work of the author. pathway may allow for combination therapies to be used to impede the emergence of resistance. However, the intracellular signal transduction pathway of FLT3 is relatively obscure. The objective of this study is to further elucidate this pathway, with particular focus on the redox signalling element which is thought to be involved. Signalling via reactive oxygen species is becoming increasingly recognised as a crucial aspect of physiological and pathological processes within the cell. The first part of this study examined the effects of NADPH oxidase-derived reactive oxygen species on the tyrosine phosphorylation levels of acute myeloid leukaemia cell lines. Using two-dimensional phosphotyrosine immunoblotting, a range of proteins were identified as undergoing tyrosine phosphorylation in response to iii NADPH oxidase activity. Ezrin, a cytoskeletal regulatory protein and substrate of Src kinase, was selected for further study.The next part of this study established that NADPH oxidase is subject to regulation by FLT3. Both wild type and oncogenic FLT3 signalling were shown to affect the expression of a key NADPH oxidase subunit, p22phox, and FLT3 was also demonstrated to drive intracellular reactive oxygen species production. The NADPH oxidase target protein, Ezrin, undergoes phosphorylation on two tyrosine residues downstream of FLT3 signalli...

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A Novel and Specific NADPH Oxidase-1 (Nox1) Small-Molecule Inhibitor Blocks the Formation of Functional Invadopodia in Human Colon Cancer Cells

Cited by 170 publications

References 40 publications

The NADPH oxidase inhibitor diphenyleneiodonium activates the human TRPA1 nociceptor

The NADPH oxidase inhibitor diphenyleneiodonium activates the human TRPA1 nociceptor

Association of Nox1 and Vinculin With Colon Cancer Progression

FLT3-driven redox-modulation of Ezrin regulates leukaemic cell migration

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