In vitro study of normoxic epidermal growth factor receptor–induced hypoxia‐inducible factor‐1‐alpha, vascular endothelial growth factor, and BNIP3 expression in head and neck squamous cell carcinoma cell lines: Implications for anti–epidermal growth factor receptor therapy

Secades, Pablo; Santa-María, Inés Saenz de; Merlo, Anna; Suárez, Carlos; Chiara, María‐Dolores

doi:10.1002/hed.23733

Cited by 12 publications

(16 citation statements)

References 35 publications

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“…Given that most tumors carrying VHL inactivating mutations are not necessarily accompanied by loss of heterozygosity, we also analyzed the impact of a VHL cancer-associated mutation, F76del VHL, on miR-210 expression by using SCC40 cells which, endogenously, express wild type VHL [18, 19]. F76del mutant was selected because represents the VHL gene alteration (c.227_229delTCT) associated with a wide spectrum of cancers including PGLs and no-neuroendocrine neoplasms related to VHL disease.…”

Section: Resultsmentioning

confidence: 99%

“…F76del mutant was selected because represents the VHL gene alteration (c.227_229delTCT) associated with a wide spectrum of cancers including PGLs and no-neuroendocrine neoplasms related to VHL disease. We first used a squamous cell carcinoma cell line, SCC40, which has been well-characterized at the genetic and functional level with regard to HIF signaling [18, 19]. SCC40 cells were transfected with a VHL -expressing construct encoding for F76del-VHL.…”

Section: Resultsmentioning

confidence: 99%

“…cDNA synthesis and real-time PCR were performed as previously described [18]. Briefly, real-time PCR reactions were performed by using SYBR Green PCR Master mix (Applied Biosystems) and the thermocycler conditions recommended by the manufacturer.…”

Section: Methodsmentioning

confidence: 99%

“…Protein extracts were obtained from SCC40 cells at 80–90% confluence as previously described [18]. Equal amounts of proteins were fractionated on SDS–PAGE and transferred to PVDF membranes.…”

Section: Methodsmentioning

confidence: 99%

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Role of VHL, HIF1A and SDH on the expression of miR-210: Implications for tumoral pseudo-hypoxic fate

et al. 2016

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The hypoxia-inducible factor 1α (HIF-1α) and its microRNA target, miR-210, are candidate tumor-drivers of metabolic reprogramming in cancer. Neuroendocrine neoplasms such as paragangliomas (PGLs) are particularly appealing for understanding the cancer metabolic adjustments because of their associations with deregulations of metabolic enzymes, such as succinate dehydrogenase (SDH), and the von Hippel Lindau (VHL) gene involved in HIF-1α stabilization. However, the role of miR-210 in the pathogenesis of SDH-related tumors remains an unmet challenge. Herein is described an in vivo genetic analysis of the role of VHL, HIF1A and SDH on miR-210 by using knockout murine models, siRNA gene silencing, and analyses of human tumors. HIF-1α knockout abolished hypoxia-induced miR-210 expression in vivo but did not alter its constitutive expression in paraganglia. Normoxic miR-210 levels substantially increased by complete, but not partial, VHL silencing in paraganglia of knockout VHL-mice and by over-expression of p76del-mutated pVHL. Similarly, VHL-mutated PGLs, not those with decreased VHL-gene/mRNA dosage, over-expressed miR-210 and accumulate HIF-1α in most tumor cells. Ablation of SDH activity in SDHD-null cell lines or reduction of the SDHD or SDHB protein levels elicited by siRNA-induced gene silencing did not induce miR-210 whereas the presence of SDH mutations in PGLs and tumor-derived cell lines was associated with mild increase of miR-210 and the presence of a heterogeneous, HIF-1α-positive and HIF-1α-negative, tumor cell population. Thus, activation of HIF-1α is likely an early event in VHL-defective PGLs directly linked to VHL mutations, but it is a late event favored but not directly triggered by SDHx mutations. This combined analysis provides insights into the mechanisms of HIF-1α/miR-210 regulation in normal and tumor tissues potentially useful for understanding the pathogenesis of cancer and other diseases sharing similar underpinnings.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Role of VHL, HIF1A and SDH on the expression of miR-210: Implications for tumoral pseudo-hypoxic fate

et al. 2016

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“…Similar to other RTKs, EGFR activation triggers multiple signal transduction pathways, including the phosphatidylinositol 3-kinase (PI3K)/AKT pathway [113] and the RAS/RAF/mitogenactivated protein kinase (MAPK) pathway [114]. Besides oxygen, ROS and particular Krebs cycle metabolites such as fumarate and succinate [115] have also been demonstrated to regulate prolyl-hydroxylase activity-mediating HIF-1α stabilization under normoxic conditions [107,116]. H 2 O 2 has been shown to induce EGF-stimulated activation of PI3K/AKT signaling in ovarian cancer cells.…”

Section: Erbb Family Receptorsmentioning

confidence: 99%

Interplay between receptor tyrosine kinases and hypoxia signaling in cancer

Glück

Aebersold

Zimmer

et al. 2015

The International Journal of Biochemistry & Cell Biology

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Chemical-agnostic hazard prediction: Statistical inference of in vitro toxicity pathways from proteomics responses to chemical mixtures

Ross

George

Bruno

et al. 2017

Computational Toxicology

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Toxicity pathways have been defined as normal cellular pathways that, when sufficiently perturbed as a consequence of chemical exposure, lead to an adverse outcome. If an exposure alters one or more normal biological pathways to an extent that leads to an adverse toxicity outcome, a significant correlation must exist between the exposure, the extent of pathway alteration, and the degree of adverse outcome. Biological pathways are regulated at multiple levels, including transcriptional, post-transcriptional, post-translational, and targeted degradation, each of which can affect the levels and extents of modification of proteins involved in the pathways. Significant alterations of toxicity pathways resulting from changes in regulation at any of these levels therefore are likely to be detectable as alterations in the proteome. We hypothesize that significant correlations between exposures, adverse outcomes, and changes in the proteome have the potential to identify putative toxicity pathways, facilitating selection of candidate targets for high throughput screening, even in the absence of a priori knowledge of either the specific pathways involved or the specific agents inducing the pathway alterations. We explored this hypothesis in vitro in BEAS-2B human airway epithelial cells exposed to different concentrations of Ni 2+ , Cd 2+ , and Cr 6+ , alone and in defined mixtures. Levels and phosphorylation status of a variety of signaling pathway proteins and cytokines were measured after 48 hours exposure, together with cytotoxicity. Least Absolute Shrinkage and Selection Operator (LASSO) multiple regression was used to identify a subset of these proteins that constitute a putative toxicity pathway capable of predicting cytotoxicity. The putative toxicity pathway for cytotoxicity of these metals and metal mixtures identified by LASSO is composed of phospho-RPS6KB1, phospho-p53, cleaved CASP3, phospho-MAPK8, IL-10, and Hif-1α. As this approach does not depend on knowledge of the chemical composition of the mixtures, it may be generally useful for identifying sets of proteins predictive of adverse effects for a variety of mixtures, including complex environmental mixtures of unknown composition.

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In vitro study of normoxic epidermal growth factor receptor–induced hypoxia‐inducible factor‐1‐alpha, vascular endothelial growth factor, and BNIP3 expression in head and neck squamous cell carcinoma cell lines: Implications for anti–epidermal growth factor receptor therapy

Abstract: The search for molecular predictors of sensitivity to gefitinib in HNSCC should be extended to the activation status of EGFR-downstream pathways, phosphorylated protein kinase B, pMAPK, and HIF-1α.

Cited by 12 publications

References 35 publications

Role of VHL, HIF1A and SDH on the expression of miR-210: Implications for tumoral pseudo-hypoxic fate

Role of VHL, HIF1A and SDH on the expression of miR-210: Implications for tumoral pseudo-hypoxic fate

Interplay between receptor tyrosine kinases and hypoxia signaling in cancer

Chemical-agnostic hazard prediction: Statistical inference of in vitro toxicity pathways from proteomics responses to chemical mixtures

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