Role of the Mevalonate Pathway in Adrenocortical Tumorigenesis

2020 ◽  
Author(s):  
Helena Panteliou Lima-Valassi ◽  
Antonio Marcondes Lerario ◽  
Luciana Ribeiro Montenegro ◽  
Maria Candida Barisson Villares Fragoso ◽  
Madson Queiroz Almeida ◽  
...  
Keyword(s):  
Cell Viability ◽  
Mevalonate Pathway ◽  
Colorimetric Assay ◽  
Rt Pcr ◽  
Expression Of Genes ◽  
Pathway Inhibition ◽  
Key Steps ◽  
Cell Growth And Proliferation ◽  
Rate Limiting

Abstract3-Hydroxy-3-methylglutaryl coenzyme A reductase (HMGCR) is the rate-limiting enzyme of the mevalonate pathway, which generates cholesterol and non-sterol compounds such as isoprenoid, which are involved in key steps of tumorigenesis such as cell growth and proliferation. Our aim was to evaluate the role of the mevalonate pathway in adrenocortical tumors (ACTs). Expression pattern of HMGCR, FDFT1, LDLR, SCARB1, StAR, TSPO, CYP11A1, CYP11B1, CYP17A1, CYP21A1, and HSD3B1 genes, involved in the mevalonate pathway and steroidogenesis, was quantified by real-time RT-PCR in 46 ACT [14 adenomas (ACA) and 11 carcinomas (ACC) from adults and 13 ACA and 8 ACC from pediatric patients]. Effects of the mevalonate pathway inhibition on NCI-H295A cell viability was assessed by colorimetric assay. HMGCR was overexpressed in most adult ACT. The expression of TSPO, STAR, CYP11B1, CYP21A1, and HSD3B1 in adult ACC was significantly lower than in ACA (p<0.05). Regarding pediatric ACT, the expression of genes involved in steroidogenesis was not different between ACA and ACC. Inhibition of isoprenoid production significantly decreased the viability of NCI-H295A cells (p<0.05). However, cholesterol synthesis blockage did not show the same effect on cell viability. Low expression of TSPO , StAR, CYP11B1, CYP21A1, and HSD3B1 characterized a signature of adult ACCs. Our data suggest that HMGCR overexpression in adult ACC might lead to intracellular isoprenoid accumulation and cell proliferation. Therefore, the mevalonate pathway is a potential target for ACC treatment.

scholarly journals Effects of Titanium Surfaces on the Developmental Profile of Monocytes/Macrophages

2014 ◽  
Vol 25 (2) ◽  
pp. 96-103 ◽  
Author(s):  
Camilla Christian Gomes Moura ◽  
Darceny Zanetta-Barbosa ◽  
Paula Dechichi ◽  
Valessa Florindo Carvalho ◽  
Priscilla Barbosa Ferreira Soares
Keyword(s):  
Cell Viability ◽  
Colorimetric Assay ◽  
Initial Period ◽  
Critical Role ◽  
Surface Characteristics ◽  
Th2 Cytokine ◽  
Titanium Surfaces ◽  
Control Surfaces ◽  
Immunoenzymatic Assays

Due to the critical role of monocytes/macrophages (Mϕ) in bone healing, this study evaluated the effects of bio-anodized, acid-etched, and machined titanium surfaces (Ti) on Mϕ behavior. Cells were separated from whole human blood from 10 patients, plated on Ti or polystyrene (control) surfaces, and cultured for 72 h. At 24, 48 and 72 h, cell viability, levels of IL1β, IL10, TNFα, TGFβ1 inflammatory mediators, and nitric oxide (NO) release were analyzed by mitochondrial colorimetric assay (MTT assay) and immunoenzymatic assays, respectively. Real-time PCR was used to verify the expression of TNFα and IL10 at 72 h. The data were subjected to a Kruskal-Wallis analysis. IL1β, TNFα and TGFβ1 release were not significantly different between the Ti surfaces (p>0.05). The presence of NO and IL10 was not detected in the samples. Cell viability did not differ between the samples cultivated on Ti and those cultivated on control surfaces, except at 24 h (p=0.0033). With respect to the mediators evaluated, the surface characteristics did not induce a typical Th1 or Th2 cytokine profile, although the cell morphology and topography were influenced by the Ti surface during the initial period.

Altering potato isoprenoid metabolism increases biomass and induces early flowering

2020 ◽  
Vol 71 (14) ◽  
pp. 4109-4124
Author(s):  
Moehninsi ◽  
Iris Lange ◽  
B Markus Lange ◽  
Duroy A Navarre
Keyword(s):  
Metabolic Regulation ◽  
Transcriptional Control ◽  
Mevalonate Pathway ◽  
Early Flowering ◽  
Bioenergy Crops ◽  
Isoprenoid Metabolism ◽  
Transgenic Lines ◽  
Coa Reductase ◽  
Rate Limiting

Abstract Isoprenoids constitute the largest class of plant natural products and have diverse biological functions including in plant growth and development. In potato (Solanum tuberosum), the regulatory mechanism underlying the biosynthesis of isoprenoids through the mevalonate pathway is unclear. We assessed the role of 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) homologs in potato development and in the metabolic regulation of isoprenoid biosynthesis by generating transgenic lines with down-regulated expression (RNAi-hmgr) or overexpression (OE) of one (StHMGR1 or StHMGR3) or two genes, HMGR and farnesyl diphosphate synthase (FPS; StHMGR1/StFPS1 or StHMGR3/StFPS1). Levels of sterols, steroidal glycoalkaloids (SGAs), and plastidial isoprenoids were elevated in the OE-HMGR1, OE-HMGR1/FPS1, and OE-HMGR3/FPS1 lines, and these plants exhibited early flowering, increased stem height, increased biomass, and increased total tuber weight. However, OE-HMGR3 lines showed dwarfism and had the highest sterol amounts, but without an increase in SGA levels, supporting a rate-limiting role for HMGR3 in the accumulation of sterols. Potato RNAi-hmgr lines showed inhibited growth and reduced cytosolic isoprenoid levels. We also determined the relative importance of transcriptional control at regulatory points of isoprenoid precursor biosynthesis by assessing gene–metabolite correlations. These findings provide novel insights into specific end-products of the sterol pathway and could be important for crop yield and bioenergy crops.

scholarly journals 3-Iodothyronamine and 3,5,3′-triiodo-L-thyronine reduce SIRT1 protein expression in the HepG2 cell line

2020 ◽  
Vol 41 (1) ◽  
Author(s):  
Ginevra Sacripanti ◽  
Leonardo Lorenzini ◽  
Lavinia Bandini ◽  
Sabina Frascarelli ◽  
Riccardo Zucchi ◽  
...  
Keyword(s):  
Protein Expression ◽  
Cell Viability ◽  
Cell Line ◽  
Potential Role ◽  
Colorimetric Assay ◽  
Tumor Cell Line ◽  
Rat Hepatocytes ◽  
Carcinoma Cells ◽  
Primary Rat Hepatocytes

AbstractBackground3-Iodothyronamine (T1AM) is an endogenous messenger chemically related to thyroid hormone. Recent results indicate significant transcriptional effects of chronic T1AM administration involving the protein family of sirtuins, which regulate important metabolic pathways and tumor progression. Therefore, the aim of this work was to compare the effect of exogenous T1AM and 3,5,3′-triiodo-L-thyronine (T3) chronic treatment on mammalian sirtuin expression in hepatocellular carcinoma cells (HepG2) and in primary rat hepatocytes at micromolar concentrations.Materials and methodsSirtuin (SIRT) activity and expression were determined using a colorimetric assay and Western blot analysis, respectively, in cells treated for 24 h with 1–20 μM T1AM or T3. In addition, cell viability was evaluated by the MTTtest upon 24 h of treatment with 0.1–20 μM T1AM or T3.ResultsIn HepG2, T1AM significantly reduced SIRT 1 (20 μM) and SIRT4 (10–20 μM) protein expression, while T3 strongly decreased the expression of SIRT1 (20 μM) and SIRT2 (any tested concentration). In primary rat hepatocytes, T3 decreased SIRT2 expression and cellular nicotinamide adenine dinucleotide (NAD) concentration, while on sirtuin activity it showed opposite effects, depending on the evaluated cell fraction. The extent of MTT staining was moderately but significantly reduced by T1AM, particularly in HepG2 cells, whereas T3 reduced cell viability only in the tumor cell line.ConclusionsT1AM and T3 downregulated the expression of sirtuins, mainly SIRT1, in hepatocytes, albeit in different ways. Differences in mechanisms are only observational, and further investigations are required to highlight the potential role of T1AM and T3 in modulating sirtuin expression and, therefore, in regulating cell cycle or tumorigenesis.

scholarly journals Induction of Macrophage M2b/c Polarization by Adipose Tissue-Derived Mesenchymal Stem Cells

2019 ◽  
Vol 2019 ◽  
pp. 1-12 ◽  
Author(s):  
Meng Sun ◽  
Linxiao Sun ◽  
Chongchu Huang ◽  
Bi-cheng Chen ◽  
Zhenxu Zhou
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Cell Viability ◽  
Culture System ◽  
Culture Medium ◽  
Peritoneal Macrophages ◽  
Rt Pcr ◽  
Control Groups ◽  
Detailed Mechanism

Background. Adipose-derived mesenchymal stem cells (ADMSCs) can promote healing and inhibit inflammation/immune response in local tissues, while the detailed mechanism remains unknown. Results. ADMSCs and peritoneal macrophages were collected from C57BL/6 mice. The culture medium (CM) from ADMSCs (24 hours cultured) was collected. The CM was added to the Mφ culture system with lipopolysaccharide (LPS) or IL-4/IL-13 or blank. And those Mφ cultures without adding CM were used as controls. A series of classification markers and signaling pathways for Mφ polarization were detected by using flow cytometry, RT-PCR, and western blotting. Furthermore, the cell viability of all the groups was detected by CCK8 assay. After CM induction in different groups, M1-Mφ markers and M2a-Mφ were decreased; however, M2b/c-Mφ markers increased. STAT3/SOCS3 and STAT6/IRF4 were suppressed in all 3 CM-treated groups. Moreover, the cell viability of all 3 groups which were induced by CM significantly increased as compared to that of the control groups without adding CM. Conclusion. ADMSCs can induce nonactivated macrophage and M1-Mφ into M2b/c-Mφ. Downregulation of the STAT3 and STAT6 pathway may involve in this process. This data shows that the anti-inflammatory role of ADMSC in local tissues may be partly due to their effect on Mφ to M2b/c-Mφ.

scholarly journals Dependence of expression of genes controlling calcium signaling on gravistimulation in cells of tomato apical leaves and treatment by etephon

2021 ◽  
Vol 65 (4) ◽  
pp. 456-465
Author(s):  
S. V. Sukhaveyeva ◽  
A. М. Kаbachevskaya ◽  
I. D. Volotovski
Keyword(s):  
Signal Transduction ◽  
Calcium Signaling ◽  
Calcium Metabolism ◽  
Early Response ◽  
Calcium Signal ◽  
Rt Pcr ◽  
Expression Of Genes ◽  
Genes Encoding ◽  
Exogenous Ethylene

The sensitivity of expression at the level of transcription of genes encoding proteins involved in calcium signal transduction to gravistimulation was revealed using real-time RT-PCR. For three genes SCA2, РВР2, САМ2, the increase in the transcript formation was shown at early response stages, starting from 15–60 minute gravistimulus. The treatment of plants before the start of gravistimulation with an ethephon (source of exogenous ethylene) led to a change in the modulation of expression of the studied genes in response to gravistimulus. The role of calcium metabolism in realization of final steps of gravitropism reaction is considered.

scholarly journals Glucose restriction drives spatial re-organization of mevalonate metabolism and liquid-crystalline lipid droplet biogenesis

2020 ◽  
Author(s):  
Sean Rogers ◽  
Hanaa Hariri ◽  
Long Gui ◽  
N. Ezgi Wood ◽  
Natalie Speer ◽  
...  
Keyword(s):  
Liquid Crystalline ◽  
Mevalonate Pathway ◽  
Dependent Manner ◽  
Metabolic Remodeling ◽  
Ester Biosynthesis ◽  
Metabolic Reactions ◽  
Glucose Restriction ◽  
Rate Limiting ◽  
Pathway Flux

AbstractEukaryotes compartmentalize metabolic pathways into sub-cellular domains, but the role of inter-organelle contacts in organizing metabolic reactions remains poorly understood. Here, we show that in response to acute glucose restriction (AGR) yeast undergo metabolic remodeling of their mevalonate pathway that is spatially coordinated at nucleus-vacuole junctions (NVJs). The NVJ serves as a metabolic platform by selectively retaining HMG-CoA Reductases (HMGCRs), driving mevalonate pathway flux in an Upc2-dependent manner. AGR-induced HMGCR compartmentalization enhances mevalonate metabolism and sterol-ester biosynthesis that generates lipid droplets (LDs) with liquid-crystalline sub-architecture. Loss of NVJ-dependent HMGCR partitioning affects yeast growth, but can be bypassed by artificially multimerizing HMGCRs, indicating NVJ compartmentalization enhances mevalonate pathway flux by promoting HMGCR inter-enzyme associations. We propose a non-canonical mechanism regulating mevalonate metabolism via the spatial compartmentalization of rate-limiting HMGCR enzymes, and reveal that AGR creates LDs with remarkable phase transition properties.One Sentence SummarySpatial compartmentalization of HMG-CoA Reductases at ER-lysosome contacts modulates mevalonate pathway flux

scholarly journals Progestins Activate the AKT Pathway in Leiomyoma Cells and Promote Survival

2009 ◽  
Vol 23 (4) ◽  
pp. 585-585 ◽  
Author(s):  
Anna V. Hoekstra ◽  
Elizabeth C. Sefton ◽  
Emily Berry ◽  
Zhenxiao Lu ◽  
Jennifer Hardt ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Cell Viability ◽  
Pi3k Inhibitor ◽  
Akt Pathway ◽  
Cell Numbers ◽  
Downstream Effectors ◽  
Proliferation And Apoptosis ◽  
Pathway Inhibition ◽  
Induced Apoptosis

ABSTRACT Context Progesterone has been associated with promoting growth of uterine leiomyomas. The mechanisms involved remain unclear. Objective In this study we investigated the activation of the AKT pathway and its downstream effectors, GSK3b and FOXO1 by progesterone as a mechanism of proliferation and survival of leiomyoma cells. Inhibitors of the AKT pathway were used to demonstrate the role of PI3K, AKT and FOXO1 in contributing to cell proliferation and apoptosis. Results Treatment of leiomyoma cells with R5020 over a period of 72h resulted in higher cell numbers compared to untreated cells. When cells were treated with 100nM R5020 for 1h and 24h, the levels of phospho(Ser 473)-AKT increased. This increase was inhibited when cells were co-treated with RU486. Treatment of leiomyoma cells with a PI3K inhibitor, LY294 dramatically decreased levels of phospho(Ser 473)-AKT, despite R5020 treatment. In addition to increased phospho(Ser 473)-AKT levels, R5020 treatment resulted in an increase in phospho(Ser 256)-FOXO1 and phospho-GSK3b. Inhibition of AKT using API-59 decreased proliferation and cell viability even in the presence of R5020. Higher concentrations of API-59 induced apoptosis of leiomyoma cells even in the presence of R5020. Psammaplysene A increased nuclear FOXO1 levels and did not affect cell proliferation but induced apoptosis of leiomyoma cells. Conclusions The progestin, R5020, can rapidly activate the AKT pathway. Inhibition of the AKT pathway inhibits cell proliferation and promotes apoptosis of leiomyoma cells.

scholarly journals Glucose restriction drives spatial reorganization of mevalonate metabolism

eLife ◽  
2021 ◽  
Vol 10 ◽  
Author(s):  
Sean M Rogers ◽  
Hanaa Hariri ◽  
N Ezgi M Wood ◽  
Natalie Ortiz Speer ◽  
W Mike Henne
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Mevalonate Pathway ◽  
Dependent Manner ◽  
Metabolic Remodeling ◽  
Spatial Reorganization ◽  
Glucose Restriction ◽  
Rate Limiting ◽  
Pathway Flux

Eukaryotes compartmentalize metabolic pathways into sub-cellular domains, but the role of inter-organelle contacts in organizing metabolic reactions remains poorly understood. Here, we show that in response to acute glucose restriction (AGR) yeast undergo metabolic remodeling of their mevalonate pathway that is spatially coordinated at nucleus-vacuole junctions (NVJs). The NVJ serves as a metabolic platform by selectively retaining HMG-CoA Reductases (HMGCRs), driving mevalonate pathway flux in an Upc2-dependent manner. Both spatial retention of HMGCRs and increased mevalonate pathway flux during AGR is dependent on NVJ tether Nvj1. Furthermore, we demonstrate that HMGCRs associate into high molecular weight assemblies during AGR in an Nvj1-dependent manner. Loss of Nvj1-mediated HMGCR partitioning can be bypassed by artificially multimerizing HMGCRs, indicating NVJ compartmentalization enhances mevalonate pathway flux by promoting the association of HMGCRs in high molecular weight assemblies. Loss of HMGCR compartmentalization perturbs yeast growth following glucose starvation, indicating it promotes adaptive metabolic remodeling. Collectively we propose a non-canonical mechanism regulating mevalonate metabolism via the spatial compartmentalization of rate-limiting HMGCR enzymes at an inter-organelle contact site.

Monitoring the marine environment for small round structured viruses (SRSVS): a new approach to combating the transmission of these viruses by molluscan shellfish

1998 ◽  
Vol 38 (12) ◽  
pp. 51-56 ◽  
Author(s):  
K. Henshilwood ◽  
J. Green ◽  
D. N. Lees
Keyword(s):  
Enteric Virus ◽  
Winter Period ◽  
Rt Pcr ◽  
Cloning And Sequencing ◽  
New Approach ◽  
Human Enteric Virus ◽  
Different Strains ◽  
The Uk ◽  
Public Health Protection

This study investigates human enteric virus contamination of a shellfish harvesting area. Samples were analysed over a 14-month period for Small Round Structured Viruses (SRSVs) using a previously developed nested RT-PCR. A clear seasonal difference was observed with the largest numbers of positive samples obtained during the winter period (October to March). This data concurs with the known winter association of gastroenteric illness due to oyster consumption in the UK and also with the majority of the outbreaks associated with shellfish harvested from this area during the study period. RT-PCR positive amplicons were further characterised by cloning and sequencing. Sequence analysis of the positive samples identified eleven SRSV strains, of both Genogroup I and Genogroup II, occurring throughout the study period. Many shellfish samples contained a mixture of strains with a few samples containing up to three different strains with both Genogroups represented. The observed common occurrence of strain mixtures may have implications for the role of shellfish as a vector for dissemination of SRSV strains. These results show that nested RT-PCR can identify SRSV contamination in shellfish harvesting areas. Virus monitoring of shellfish harvesting areas by specialist laboratories using RT-PCR is a possible approach to combating the transmission of SRSVs by molluscan shellfish and could potentially offer significantly enhanced levels of public health protection.

New trends in the pharmacological intervention of PPARs in obesity: Role of natural and synthetic compounds_

2020 ◽  
Vol 28 ◽  
Author(s):  
Seyed Mohammad Nabavi ◽  
Kasi Pandima Devi ◽  
Sethuraman Sathya ◽  
Ana Sanches-Silva ◽  
Listos Joanna ◽  
...  
Keyword(s):  
Tissue Expression ◽  
Health Concern ◽  
Pharmacological Intervention ◽  
Peroxisome Proliferator ◽  
Expression Of Genes ◽  
Ppar Agonists ◽  
Prevalence Of Obesity ◽  
Peroxisome Proliferator Activated Receptors ◽  
Natural And Synthetic

: Obesity is a major health concern for a growing fraction of the population, with the prevalence of obesity and its related metabolic disorders not being fully understood. Over the last decade, many attempts have been undertaken to understand the mechanisms at the basis of this condition, in which the accumulation of fat occurring in adipose tissue, leads to the pathogenesis of obesity related disorders. Among the most recent studies, those on Peroxisome Proliferator Activated Receptors (PPARs) revealed that these nuclear receptor proteins acting as transcription factors, among others, regulate the expression of genes involved in energy, lipid, and glucose metabolisms, and chronic inflammation. The three different isotypes of PPARs, with different tissue expression and ligand binding specificity, exert similar or overlapping functions directly or indirectly linked to obesity. In this study, we reviewed the available scientific reports concerning the PPARs structure and functions, especially in obesity, considering both natural and synthetic ligands and their role in the therapy of obesity and obesity-associated disorders. In the whole, the collected data show that there are both natural and synthetic compounds that show beneficial promising activity as PPAR agonists in chronic diseases related to obesity.

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