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Cells exposed to hypoxia respond by increasing the level of hypoxia-inducible factor-1 (HIF-1). This factor then activates a number of genes by binding to hypoxia response elements in their promoter regions. A second hypoxia-responsive factor, HIF-2, can activate many of the same genes as HIF-1. Overexpression of HIFs accompanies the pathogenesis o...
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... aliquot of the corresponding samples diluted 1:10 was used in the real-time PCR experiments (27). The cDNA were amplified in TaqMan universal PCR master mix by the ABI Prism 7000 sequence detection system (Applied Biosystems) with primers and probe sets as shown in Table 2. Human h-actin endogenous control primers and probe set were provided by Applied Biosystems. ...
Similar publications
Hypoxia-inducible factor (HIF) is an attractive therapeutic target for renal cell carcinoma (RCC) as its high expression due to the loss of von Hippel-Lindau (VHL) promotes RCC progression. Considering this, we hypothesized that ELR510444, a novel orally available small molecule inhibitor of HIF activity, would reduce angiogenesis and possess signi...
The transcriptional activator complex HIF-1 plays a key role in the long term adaptation of cells and tissues to their hypoxic microenvironment by stimulating the expression of genes involved in angiogenesis and glycolysis. The expression of the HIF-1 complex is regulated by the levels of its HIF-alpha subunits that are degraded under normoxic cond...
Hypoxia is an imbalance between oxygen supply and demand, which deprives cells or tissues of sufficient oxygen. It is well-established that hypoxia triggers adaptive responses, which contribute to short- and long-term pathologies such as inflammation, cardiovascular disease and cancer. Induced by both microenvironmental hypoxia and genetic mutation...
Citations
... Glucose uptake was enhanced in the hematopoietic stem cells (HSC) expressing STAT5, and HIF-2α is required for the upregulation of genes associated with glucose metabolism; in T cells, it also has been observed that STAT5 mediated the glucose uptake [18]. Both isoforms of HIF, HIF-1α and HIF-2α, regulate the expression of numerous common genes, while HIF-1α induces preferentially genes of the glycolytic pathway [19,20]. ...
Citation: Valle-Mendiola, A.; Rocha-Zavaleta, L.; Maldonado-Lagunas, V.; Morelos-Laguna, D.; Gutiérrez-Hoya, A.; Weiss-Steider, B.; Soto-Cruz, I. STAT5 Is Necessary for the Metabolic Switch Induced by IL-2 in Cervical Cancer Cell Line SiHa. Int. J. Mol. Sci. 2024, 25, 6835. https:// Abstract: The tumor cells reprogram their metabolism to cover their high bioenergetic demands for maintaining uncontrolled growth. This response can be mediated by cytokines such as IL-2, which binds to its receptor and activates the JAK/STAT pathway. Some reports show a correlation between the JAK/STAT pathway and cellular metabolism, since the constitutive activation of STAT proteins promotes glycolysis through the transcriptional activation of genes related to energetic metabolism. However, the role of STAT proteins in the metabolic switch induced by cytokines in cervical cancer remains poorly understood. In this study, we analyzed the effect of IL-2 on the metabolic switch and the role of STAT5 in this response. Our results show that IL-2 induces cervical cancer cell proliferation and the tyrosine phosphorylation of STAT5. Also, it induces an increase in lactate secretion and the ratio of NAD+/NADH, which suggest a metabolic reprogramming of their metabolism. When STAT5 was silenced, the lactate secretion and the NAD+/NADH ratio decreased. Also, the expression of HIF1α and GLUT1 decreased. These results indicate that STAT5 regulates IL-2-induced cell proliferation and the metabolic shift to aerobic glycolysis by regulating genes related to energy metabolism. Our results suggest that STAT proteins modulate the metabolic switch in cervical cancer cells to attend to their high demand of energy required for cell growth and proliferation.
... A lack of HIF-1ɑ protein expression in articular cartilage leads to marked chondrocyte death. Expression of HIF-1ɑ is upregulated in a hypoxic environment, leading to the activation of downstream genes such as IL-1β, TNF-ɑ, VEGF and NOS2, further exacerbating osteoarthritis pathology [34]. Importantly, NO was reported not only to modulate the HIF-1 response under hypoxic conditions, but to also function as an inducer of HIF-1. ...
Modified Jiawei Juanbi decoction (MJD) is used for the treatment of early-stage knee osteoarthritis (KOA). Here, modified Jiawei Juanbi decoction (MJD) was employed for the treatment of early-stage knee osteoarthritis (KOA) and its mechanisms were assessed via metabonomics and network pharmacology. A total of 24 male Sprague-Dawley rats were randomly allocated into a normal control group, a model group, and an MJD group (n = 8 rats per group). Each rat group was further equally divided into two subgroups for investigation for either 14 or 28 days. A rat model of early-stage KOA was constructed and rats were treated with MJD. Effects were evaluated based on changes in knee circumference, mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL). We also analyzed histopathological changes in articular cartilage. High-resolution mass spectrometry was used to analyze the chemical profile of MJD, identifying 228 components. Using an LC-Q-TOF-MS metabonomics approach, 33 differential metabolites were identified. The relevant pathways significantly associated with MJD include arginine and proline metabolism, vitamin B6 metabolism, as well as the biosynthesis of phenylalanine, tyrosine and tryptophan. The system pharmacology paradigm revealed that MJD contains 1027 components and associates with 1637 genes, of which 862 disease genes are related to osteoarthritis. The construction of the MJD composition-target-KOA network revealed a total of 140 intersection genes. A total of 39 hub genes were identified via integration of betweenness centrality values greater than 100 using CytoHubba. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis revealed several significantly affected signaling pathways including the HIF-1, AGE-RAGE (in diabetic complications), IL-17, rheumatoid arthritis and TNF pathways. Integrated-omics and network pharmacology approaches revealed a necessity for further detailed investigation focusing on two major targets, namely NOS2 and NOS3, along with their essential metabolite (arginine) and associated pathways (HIF-1 signaling and arginine and proline metabolism). Real-time PCR validated significantly greater downregulation of NOS2 and HIF-1ɑ in the MJD as compared to the model group. Molecular docking analysis further confirmed the binding of active MJD with key active components. Our findings elucidate the impact of MJD on relevant pathophysiological and metabolic networks relevant to KOA and assess the drug efficacy of MJD and its underlying mechanisms of action.
... The first is hypoxia-inducible factors one and two α (HIF1α and HIF2α). HIF1α is known to slightly upregulate PKIB, and HIF2α strongly upregulates (greater than 2-fold increase) both PKIB and PKIα [42]. Interestingly, while HIF1α does upregulate PKIB, it does not upregulate PKIα, despite the strong similarity between the PKIα and PKIB genes [42]. ...
... HIF1α is known to slightly upregulate PKIB, and HIF2α strongly upregulates (greater than 2-fold increase) both PKIB and PKIα [42]. Interestingly, while HIF1α does upregulate PKIB, it does not upregulate PKIα, despite the strong similarity between the PKIα and PKIB genes [42]. Further research into the link between PKIB and HIF1α demonstrated that the PKIB promoter is bound to HIF1α in beta cells so that increasing amounts of HIF1α show a corresponding increase in the expression of PKIB ( Figure 2C) [43]. ...
The serine-threonine kinase protein kinase A (PKA) is a cyclic AMP (cAMP)-dependent intracellular protein with multiple roles in cellular biology including metabolic and transcription regulation functions. The cAMP-dependent protein kinase inhibitor β (PKIB) is one of three known endogenous protein kinase inhibitors of PKA. The role of PKIB is not yet fully understood. Hormonal signaling is correlated with increased PKIB expression through genetic regulation, and increasing PKIB expression is associated with decreased cancer patient prognosis. Additionally, PKIB impacts cancer cell behavior through two mechanisms; the first is the nuclear modulation of transcriptional activation and the second is the regulation of oncogenic AKT signaling. The limited research into PKIB indicates the oncogenic potential of PKIB in various cancers. However, some studies suggest a role of PKIB in non-cancerous disease states. This review aims to summarize the current literature and background of PKIB regarding cancer and related issues. In particular, we will focus on cancer development and therapeutic possibilities, which are of paramount interest in PKIB oncology research.
... To evaluate the transcriptional effects of this EPAS1 variant in response to hypoxia, isogenic wild-type, heterozygous, and homozygous clones were cultured under conditions of normoxia (21% O 2 ) and hypoxia (1% O 2 ) for 12 and 24 hours. The selection of these conditions were based on previous literature in which these O 2 tensions and time points were implemented to study HIF activity in HEK293T cells (49)(50)(51)(52). ...
... The use of a cell culture model system with O 2 exposure ranging from room air to as low as 0.5% provides a powerful tool to study cellular mechanisms in hypoxia. The selection of 1% O 2 was based off prior studies in which O 2 tension induced HIF-2 stabilization in HEK293T cells (49)(50)(51)(52). O 2 and CO 2 levels were calibrated before experimentation against external sensors. ...
Hypoxia-inducible factor pathway genes are linked to adaptation in both human and nonhuman highland species. EPAS1 , a notable target of hypoxia adaptation, is associated with relatively lower hemoglobin concentration in Tibetans. We provide evidence for an association between an adaptive EPAS1 variant (rs570553380) and the same phenotype of relatively low hematocrit in Andean highlanders. This Andean-specific missense variant is present at a modest frequency in Andeans and absent in other human populations and vertebrate species except the coelacanth. CRISPR-base-edited human cells with this variant exhibit shifts in hypoxia-regulated gene expression, while metabolomic analyses reveal both genotype and phenotype associations and validation in a lowland population. Although this genocopy of relatively lower hematocrit in Andean highlanders parallels well-replicated findings in Tibetans, it likely involves distinct pathway responses based on a protein-coding versus noncoding variants, respectively. These findings illuminate how unique variants at EPAS1 contribute to the same phenotype in Tibetans and a subset of Andean highlanders despite distinct evolutionary trajectories.
... Note that we observed two non-specific bands that were not lost when LOX was depleted, and which were apparent in some, but not all western blots. Next, we established our assay conditions and show that, in agreement with previous studies [31][32][33] , LOX was strongly induced by hypoxia (1% O 2 for 24 h) in MDA-MB-231 cells ( Supplementary Fig. 2b). Notably, oxygen consumption increases at higher cell densities 34 , and accordingly we show that LOX expression increased when MDA-MB-231 cells were grown at higher cell density ( Supplementary Fig. 2b). ...
Congenital disorders of glycosylation (CDG) are rare genetic disorders with a spectrum of clinical manifestations caused by abnormal N-glycosylation of secreted and cell surface proteins. Over 130 genes are implicated and next generation sequencing further identifies potential disease drivers in affected individuals. However, functional testing of these variants is challenging, making it difficult to distinguish pathogenic from non-pathogenic events. Using proximity labelling, we identified OST48 as a protein that transiently interacts with lysyl oxidase (LOX), a secreted enzyme that cross-links the fibrous extracellular matrix. OST48 is a non-catalytic component of the oligosaccharyltransferase (OST) complex, which transfers glycans to substrate proteins. OST48 is encoded by DDOST, and 43 variants of DDOST are described in CDG patients, of which 34 are classified as variants of uncertain clinical significance (VUS). We developed an assay based on LOX N-glycosylation that confirmed two previously characterised DDOST variants as pathogenic. Notably, 39 of the 41 remaining variants did not have impaired activity, but we demonstrated that p.S243F and p.E286del were functionally impaired, consistent with a role in driving CDG in those patients. Thus, we describe a rapid assay for functional testing of clinically relevant CDG variants to complement genome sequencing and support clinical diagnosis of affected individuals.
... Besides, it has been suggested that LINC02015 is possibly related to vascular injury, calcium ion regulation, and cell proliferation, which are important in a lot of cardiovascular diseases [14,15]. What's more, through online Gene Expression Omnibus (GEO) profiles, we find different expression levels of LINC02015 in cardiovascular relating situations, including tobacco use and hypoxia, which could damage our arteries [16,17]. However, there is no clue whether LINC02015 affects the cell behaviors of VSMCs, which are crucial in the pathogenesis of various aortic diseases. ...
Long intergenic nonprotein coding RNA 2015 (LINC02015) is a long non-coding RNA that has been found elevated in various cell proliferation-related diseases. However, the functions and interactive mechanism of LINC02015 remain unknown. This study aimed to explore the role of LINC02015 in the cell proliferation and apoptosis of vascular smooth muscle cells (VSMCs) to explain the pathogenesis of aortic diseases. Ascending aorta samples and angiotensin-II (AT-II) treated primary human aortic VSMCs (HAVSMCs) were used to evaluate the LINC02015 expression. RNA sequencing, chromatin isolation by RNA purification sequencing, RNA pull-down, and mass spectrometry (MS) were applied to explore the potential interacting mechanisms. LINC02015 expression was found elevated in aortic dissection and AT-II-treated HAVSMCs. Cell proliferation and cell cycle were activated in HAVSMCs with LINC02015 knockdown. The cyclins family and caspase family were found to participate in regulating the cell cycle and apoptosis via the NF-κB signaling pathway. RXRA was discovered as a possible hub gene for LINC02015 transcriptional regulating networks. Besides, the protein interaction network of LINC02015 was revealed with candidate regulating molecules. It was concluded that the knockdown of LINC02015 could promote cell proliferation and inhibit the apoptosis of HAVSMCs through an RXRA-related transcriptional regulation network, which could provide a potential therapeutic target for aortic diseases.
... The signaling and/or the transcription factors involved in these effects are not known to date. PTPRZ1 expression increased after hypoxia-inducible factor (HIF) 2 but not HIF1 vector transfection in HEK293T cells, suggesting that the Ptprz1 gene may be a target of hypoxia [17]. It was later found that the Ptprz1 promoter contains HIF-and E26 transformationspecific (Ets)-binding motifs, and the preferential activation of Ptprz1 by HIF2, but not HIF1, may derive from the cooperative binding of HIF2 and Ets Like-1 (ELK1) to the nearby corresponding sites on the Ptprz1 promoter [18]. ...
Protein tyrosine phosphatase receptor zeta 1 (PTPRZ1) is a type V transmembrane tyrosine phosphatase that is highly expressed during embryonic development, while its expression during adulthood is limited. PTPRZ1 is highly detected in the central nervous system, affecting oligoden-drocytes' survival and maturation. In gliomas, PTPRZ1 expression is significantly upregulated and is being studied as a potential cancer driver and as a target for therapy. PTPRZ1 expression is also increased in other cancer types, but there are no data on the potential functional significance of this finding. On the other hand, low PTPRZ1 expression seems to be related to a worse prognosis in some cancer types, suggesting that in some cases, it may act as a tumor-suppressor gene. These discrepancies may be due to our limited understanding of PTPRZ1 signaling and tumor microenvironments. In this review, we present evidence on the role of PTPRZ1 in angiogenesis and cancer and discuss the phenomenal differences among the different types of cancer, depending on the regulation of its tyrosine phosphatase activity or ligand binding. Clarifying the involved signaling pathways will lead to its efficient exploitation as a novel therapeutic target or as a biomarker, and the development of proper therapeutic approaches.
... The function of HIF-3α, which lacks the transactivation domain, is not clear in detail. HIF-3α has many splice variants [93]; some variants appear to activate gene expression [94,95], while others might be negative regulators of HIF-1α and HIF-2α [96][97][98]. ...
Ferroptosis is an oxidative damage-related, iron-dependent regulated cell death with intracellular lipid peroxide accumulation, which is associated with many physiological and pathological processes. It exhibits unique features that are morphologically, biochemically, and immunologically distinct from other regulated cell death forms. Ferroptosis is regulated by iron metabolism, lipid metabolism, anti-oxidant defense systems, as well as various signal pathways. Hypoxia, which is found in a group of physiological and pathological conditions, can affect multiple cellular functions by activation of the hypoxia-inducible factor (HIF) signaling and other mechanisms. Emerging evidence demonstrated that hypoxia regulates ferroptosis in certain cell types and conditions. In this review, we summarize the basic mechanisms and regulations of ferroptosis and hypoxia, as well as the regulation of ferroptosis by hypoxia in physiological and pathological conditions, which may contribute to the numerous diseases therapies.
... As such, their overexpression in 3D tumor spheroids derived from LUAD and breast cancer cell lines was somehow unexpected; however, it could be suggestive of a broad neuronal-specific gene expression reprogramming of cancer cells during detachment from the ECM and 3D tumor growth. Although still poorly defined, the literature suggests that both isoenzymes exert non-canonical "moonlighting" functions in carcinogenesis [122,123]. Under hypoxia, HIF1a binds to the hypoxia-responsive element (HRE) on the promoter region of ALDOC, thus causing metabolic reprogramming or aberration of glycolysis to promote glioblastoma and ovarian cancer [124]. ...
Background
Metastases are the major cause of cancer-related morbidity and mortality. By the time cancer cells detach from their primary site to eventually spread to distant sites, they need to acquire the ability to survive in non-adherent conditions and to proliferate within a new microenvironment in spite of stressing conditions that may severely constrain the metastatic process. In this study, we gained insight into the molecular mechanisms allowing cancer cells to survive and proliferate in an anchorage-independent manner, regardless of both tumor-intrinsic variables and nutrient culture conditions.
Methods
3D spheroids derived from lung adenocarcinoma (LUAD) and breast cancer cells were cultured in either nutrient-rich or -restricted culture conditions. A multi-omics approach, including transcriptomics, proteomics, and metabolomics, was used to explore the molecular changes underlying the transition from 2 to 3D cultures. Small interfering RNA-mediated loss of function assays were used to validate the role of the identified differentially expressed genes and proteins in H460 and HCC827 LUAD as well as in MCF7 and T47D breast cancer cell lines.
Results
We found that the transition from 2 to 3D cultures of H460 and MCF7 cells is associated with significant changes in the expression of genes and proteins involved in metabolic reprogramming. In particular, we observed that 3D tumor spheroid growth implies the overexpression of ALDOC and ENO2 glycolytic enzymes concomitant with the enhanced consumption of glucose and fructose and the enhanced production of lactate. Transfection with siRNA against both ALDOC and ENO2 determined a significant reduction in lactate production, viability and size of 3D tumor spheroids produced by H460, HCC827, MCF7, and T47D cell lines.
Conclusions
Our results show that anchorage-independent survival and growth of cancer cells are supported by changes in genes and proteins that drive glucose metabolism towards an enhanced lactate production. Notably, this finding is valid for all lung and breast cancer cell lines we have analyzed in different nutrient environmental conditions. broader Validation of this mechanism in other cancer cells of different origin will be necessary to broaden the role of ALDOC and ENO2 to other tumor types. Future in vivo studies will be necessary to assess the role of ALDOC and ENO2 in cancer metastasis.
... Homozygous EPAS1-deficient mice failed to survive in the mid-gestational stage owing to profound bradycardia and circulatory failure. Additionally, HIF-2α plays an indispensable role in angiogenesis compared to HIF-1α, which mainly activates the glucose metabolic pathways (Wang et al., 2005). Peng (Peng et al., 2000) found that EPAS1-deficient embryos failed to form large vessels or seal intact structures in the yolk sac, indicating improper vascular remodeling during vasculogenesis. ...
Pulmonary hypertension (PH) is a group of syndromes characterized by irreversible vascular remodeling and persistent elevation of pulmonary vascular resistance and pressure, leading to ultimately right heart failure and even death. Current therapeutic strategies mainly focus on symptoms alleviation by stimulating pulmonary vessel dilation. Unfortunately, the mechanism and interventional management of vascular remodeling are still yet unrevealed. Hypoxia plays a central role in the pathogenesis of PH and numerous studies have shown the relationship between PH and hypoxia-inducible factors family. EPAS1, known as hypoxia-inducible factor-2 alpha (HIF-2α), functions as a transcription factor participating in various cellular pathways. However, the detailed mechanism of EPAS1 has not been fully and systematically described. This article exhibited a comprehensive summary of EPAS1 including the molecular structure, biological function and regulatory network in PH and other relevant cardiovascular diseases, and furthermore, provided theoretical reference for the potential novel target for future PH intervention.
