Assessing Bioenergetic Function in Response to Reactive Oxygen Species in Neural Cells

2021 ◽  
Vol 11 ◽  
Author(s):  
Mahyar Sameti ◽  
Pablo R. Castello ◽  
Matthew Lanoue ◽  
Tatiana Karpova ◽  
Carlos F. Martino
Keyword(s):  
Oxidative Stress ◽  
Cell Line ◽  
Cell Lines ◽  
Oxidative Metabolism ◽  
Neuronal Cells ◽  
Flux Analysis ◽  
Neural Cells ◽  
Atp Production ◽  
Extracellular Flux ◽  
Energy Demands

In this study, we characterized the bioenergetic response of the Lund human mesencephalic (LUHMES) cell line and a mouse astrocyte cell line to oxidative stress. Extracellular hydrogen peroxide  (H2O2) levels and bioenergetic response were investigated in these cell lines after exposure to paraquat (PQ), a redox cycling compound that causes oxidative stress in cells. We used extracellular flux analysis to measure mitochondrial function in adherent astrocytes and LUHMES cells. Extracellular H2O2 was measured fluorometrically. H2O2 levels increased in both cell lines after exposure to 5 µM PQ for 18 h; however, the extent of H2O2 increase with astrocytes was significantly lower than that with LUHMES cells (33% vs. 67%). Measurements of basal mitochondrial respiration showed that PQ almost completely eliminated oxygen consumption rate (OCR) in astrocytes and significantly reduced it in LUHMES cells. Notably, OCR in LUHMES cells was higher than that in astrocytes, indicating that neuronal cells maintain higher oxidative metabolism than glial cells, which is also consistent with higher energy demands of the neuronal cells. Moreover, LUHMES cells exhibited a higher amount of adenosine triphosphate (ATP) being produced by oxidative phosphorylation than by glycolysis. In contrast, astrocytes demonstrated a higher glycolytic capacity and glycolytic reserve than LUHMES cells and higher ATP production rate by glycolysis than its production by mitochondrial oxidative metabolism. Collectively, this study showed the differential bioenergetic responses between astrocytes and LUHMES cells in responding to oxidative stress and the findings may provide insights into the mitochondrial reserve capacity in neurons and astrocytes in responding to oxidative stress. (First online: Mar 30, 2021)

scholarly journals Increased Susceptibility to Ethylmercury-Induced Mitochondrial Dysfunction in a Subset of Autism Lymphoblastoid Cell Lines

2015 ◽  
Vol 2015 ◽  
pp. 1-13 ◽  
Author(s):  
Shannon Rose ◽  
Rebecca Wynne ◽  
Richard E. Frye ◽  
Stepan Melnyk ◽  
S. Jill James
Keyword(s):  
Oxidative Stress ◽  
Mitochondrial Dysfunction ◽  
Cell Lines ◽  
Autism Spectrum ◽  
Reserve Capacity ◽  
Flux Analysis ◽  
Lymphoblastoid Cell Lines ◽  
Extracellular Flux ◽  
Respiratory Parameters ◽  
Increased Risk

The association of autism spectrum disorders with oxidative stress, redox imbalance, and mitochondrial dysfunction has become increasingly recognized. In this study, extracellular flux analysis was used to compare mitochondrial respiration in lymphoblastoid cell lines (LCLs) from individuals with autism and unaffected controls exposed to ethylmercury, an environmental toxin known to deplete glutathione and induce oxidative stress and mitochondrial dysfunction. We also tested whether pretreating the autism LCLs with N-acetyl cysteine (NAC) to increase glutathione concentrations conferred protection from ethylmercury. Examination of 16 autism/control LCL pairs revealed that a subgroup (31%) of autism LCLs exhibited a greater reduction in ATP-linked respiration, maximal respiratory capacity, and reserve capacity when exposed to ethylmercury, compared to control LCLs. These respiratory parameters were significantly elevated at baseline in the ethylmercury-sensitive autism subgroup as compared to control LCLs. NAC pretreatment of the sensitive subgroup reduced (normalized) baseline respiratory parameters and blunted the exaggerated ethylmercury-induced reserve capacity depletion. These findings suggest that the epidemiological link between environmental mercury exposure and an increased risk of developing autism may be mediated through mitochondrial dysfunction and support the notion that a subset of individuals with autism may be vulnerable to environmental influences with detrimental effects on development through mitochondrial dysfunction.

scholarly journals A Comparative Study of the Cytotoxic Effects and Oxidative Stress of Gossypol on Bovine Kidney and HeLa Cell Lines

2021 ◽  
Vol 15 (3) ◽  
pp. 157-164
Author(s):  
Mahsa Daneshmand ◽  
◽  
Jamileh Salar Amoli ◽  
Tahereh Ali Esfahani ◽  
◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Hela Cell ◽  
Cell Line ◽  
Cell Lines ◽  
Toxic Effects ◽  
Cytotoxic Effects ◽  
Bovine Kidney ◽  
Hela Cell Lines ◽  
Significant Difference ◽  
And Oxidative Stress

Background: Cotton seed is one of the main sources of protein in animal feeds, containing gossypol, which has been shown to have toxic effects. Results reported by various studies also indicate the anti-cancer effects of gossypol on various cell types. However, its toxic effects on human and animal cells have not been fully established. This study was planned to investigate, for the first time, the cytotoxic effects and oxidative stress induced by gossypol on normal Bovine Kidney (BK) and HeLa cell lines, representing typical healthy and cancer cells, respectively. Methods: The BK and HeLa cell lines were treated for 24, 48 or 72 hours with 5, 10 or 20 ppm of gossypol (+/-). The cellular bio-availability and cytotoxicity were measured by MTT assay. The catalase and Malondialdehyde (MDA) levels were also measured to represent the oxidative stress parameters. Results: The percentages of cytotoxicity in BK and HeLa cell lines were calculated at a gossypol concentration of 5, 10 and 20 ppm over 24, 48 or 72 hours of incubation, respectively. The Lethal Concentration 50 (lC50) values were also determined for the two cell lines. No changes in the catalase and lipid peroxidase activities were observed in either cell line. Conclusion: The percentage of the gossypol cytotoxicity was concentration-dependent. By comparing the IC50 in both cell lines using one-way Analysis of Variance (ANOVA) analysis, a significant difference was observed, suggesting that Hela cells were less sensitive to gossypol than the BK cells. Lack of changes in the oxidative stress, as tested by catalase and MDA assays, demonstrated that gossypol did not induce oxidative stress in either cell line.

A comparative study of using free radical generators in the testing of chosen oxidative stress parameters in the different types of cells

2018 ◽  
Vol 34 (4) ◽  
pp. 237-252 ◽  
Author(s):  
Katarzyna Miranowicz-Dzierżawska
Keyword(s):  
Oxidative Stress ◽  
Superoxide Dismutase ◽  
Free Radicals ◽  
Cell Line ◽  
Cell Lines ◽  
Chinese Hamster ◽  
Cancer Cell Line ◽  
Lung Cancer Cell Line ◽  
Superoxide Dismutase Activity ◽  
Raw 264.7

The aim of this study was to assess whether there are differences between the results of determining oxidative stress markers obtained from different origin cell lines after exposure to chemicals generating free radicals. The studies considered two markers of oxidative stress: the level of thiobarbituric acid reactive substances (TBARS) and superoxide dismutase activity. The evaluation was performed in five cell lines: Chinese hamster ovary (CHO-9) cells, lung adenocarcinoma A549, macrophages RAW264.7, skin carcinoma cells A431, and keratinocytes HaCaT. Three compounds generating free radicals were used as a source of reactive oxygen/nitrogen: 2,2′-azobis-2-methyl-propanimidamide dihydrochloride (AAPH), sodium persulfate (SP), and 3-morpholinosydnonimine hydrochloride (SIN-1). The most appropriate cell line to assess the level of TBARS proved to be the murine macrophage cell line RAW 264.7. Equally, good performance was observed in the lung cancer cell line A549, but only when tested with AAPH and SP. In the case of measuring superoxide dismutase activity, it appeared that the most suitable cell line was also the RAW 264.7 line, although dispersion increased significantly at the highest concentrations of AAPH and SP measurements. When choosing a cell line to determine oxidative stress, the specificity of the stress-inducing compound and the parameter determined should be taken into consideration.

scholarly journals Molecular tuning of the axonal mitochondrial Ca2+ uniporter ensures metabolic flexibility of neurotransmission

2019 ◽  
Author(s):  
Ghazaleh Ashrafi ◽  
Jaime de Juan-Sanz ◽  
Ryan Farrell ◽  
Timothy A Ryan
Keyword(s):  
Oxidative Metabolism ◽  
Neuronal Cells ◽  
Atp Synthesis ◽  
Specific Protein ◽  
Metabolic Flexibility ◽  
Nerve Terminals ◽  
Atp Production ◽  
Presynaptic Function ◽  
The Brain ◽  
Do So

The brain is a vulnerable metabolic organ and must adapt to different fuel conditions to sustain function. Nerve terminals are a locus of this vulnerability but how they regulate ATP synthesis as fuel conditions vary is unknown. We show that synapses can switch from glycolytic to oxidative metabolism, but to do so, they rely on activity-driven presynaptic mitochondrial Ca2+ uptake to accelerate ATP production. We demonstrate that while in non-neuronal cells mitochondrial Ca2+ uptake requires elevated extramitochondrial Ca2+, axonal mitochondria readily take up Ca2+ in response to small changes in Ca2+. We identified the brain-specific protein MICU3 as a critical driver of this tuning of Ca2+ sensitivity. Ablation of MICU3 renders axonal mitochondria similar to non-neuronal mitochondria, prevents acceleration of local ATP synthesis, and impairs presynaptic function under oxidative conditions. Thus, presynaptic mitochondria rely on MICU3 to facilitate mitochondrial Ca2+ uptake during activity and achieve metabolic flexibility.

Antileukemic Effects of the Novel Agent Elesclomol.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 2736-2736 ◽  
Author(s):  
Sue Chow ◽  
Masazumi Nagai ◽  
Suqin He ◽  
Ronald K Blackman ◽  
James Barsoum ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Membrane Potential ◽  
Cell Line ◽  
Mitochondrial Membrane Potential ◽  
Cell Lines ◽  
Research Funding ◽  
Mitochondrial Membrane ◽  
Membrane Integrity ◽  
Employment Equity ◽  
Equity Ownership

Abstract Abstract 2736 Poster Board II-712 Elesclomol (N-malonyl-bis (N′-methyl-N′-thiobenzoyl hydrazide)) is an investigational first-in-class oxidative stress inducer that triggers apoptosis in cancer cells (Kirshner et al., Mol Cancer Ther 2008;7:2319–27). In the clinic, elesclomol is well tolerated in humans and showed activity in combination with paclitaxel in patients with refractory solid tumors (Berkenblit et al., Clin Cancer Res 2007;13:584–90). The aims of the current study are to examine the activity of elesclomol against a range of AML cell lines, including primary patient blast cultures, to investigate the mechanisms of drug action and the potential to combine elesclomol with other agents, and to identify candidate biomarkers for monitoring effects during treatment of leukemia patients with elesclomol. Here we describe the effects of elesclomol treatment in 4 AML cell lines selected based on their varying molecular attributes. Effects on cellular redox state and mitochondrial function were monitored using a flow cytometry incorporating the glutathione (GSH) probe monobromobimane, the reactive oxygen species (ROS) probe carboxy-dichlorofluorescin and the mitochondrial membrane potential stain DiIC(1)5. In addition, outer cell membrane integrity was determined by propidium iodide exclusion. Dual staining of fixed, permeabilized cells with phospho-specific antibodies to p38 and SAPK/JNK was used to determine if elesclomol treatment results in activation of the stress-activated MAP kinase pathways. Elesclomol showed potent anti-leukemic effects in vitro at concentrations as low as 10nM, which is well below the concentrations achieved in cancer patients, and greater toxicity was achieved with prolonged drug exposure. In OCI-AML2, a factor-independent, poorly differentiated AML cell line, toxicity was associated with loss of reduced GSH that coincided with a large increase in ROS generation and depolarization of the mitochondrial inner membrane, and later with loss of surface membrane integrity. A similar pattern was seen in OCI-M2, a p53-deficient erythroblastic leukemia cell line, except that during the early stages of drug effect these cells showed a large increase in reduced GSH, suggesting that initially they are able to compensate for drug-induced oxidative stress through enhanced cellular antioxidant production. In contrast, the factor-dependent line OCI-AML5, which appeared most sensitive to elesclomol, showed loss of outer membrane integrity without obvious prior oxidative stress while the Flt-3 ITD mutant line MV4-11 showed an initial loss of mitochondrial membrane potential without accompanying oxidative stress. Strikingly, we did not observe activation of the stress-responsive p38 or SAPK/JNK pathways in any of these 4 cell lines tested, suggesting that this is not a prominent response to elesclomol activity in AML and that additional mechanisms may be at work for the activity of elesclomol in these cells. Further investigations are ongoing and additional studies, including evaluation of elesclomol activity in primary leukemic cells from AML patients, will be presented. In summary, elesclomol is a potent novel compound that exerts anti-leukemic effects in tissue culture at drug concentrations that are well below those achieved in patients, suggesting that it might be active in leukemia patients. Disclosures: Chow: Synta Pharmaceuticals Inc.: Research Funding. Nagai:Synta Pharmaceuticals Inc.: Employment. He:Synta Pharmaceuticals Inc.: Employment. Blackman:Synta Pharmaceuticals Inc.: Employment, Equity Ownership. Barsoum:Synta Pharmaceuticals Inc.: Employment, Equity Ownership. Vukovic:Synta Pharmaceuticals Inc.: Employment, Equity Ownership. Hedley:Synta Pharmaceuticals Inc.: Research Funding.

FOXO1-p300-Txn Circuit Regulates Oxidative Stress Responses in Diffuse Large B-Cell Lymphomas Characterized By Enhanced Oxidative Phosphorylation

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 466-466 ◽  
Author(s):  
Tomasz Sewastianik ◽  
Maciej Szydlowski ◽  
Emilia Bialopiotrowicz ◽  
Ewa Jablonska ◽  
Przemyslaw Kiliszek ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Oxidative Phosphorylation ◽  
B Cell ◽  
Mrna Expression ◽  
Cell Line ◽  
Cell Lines ◽  
Mrna Abundance ◽  
Repressor Activity ◽  
Dlbcl Subtype ◽  
Large B Cell

Abstract Diffuse large B-cell lymphoma (DLBCL) is a clinically and molecularly heterogeneous disease. The comparison of DLBCLs transcriptional profiles using multiple clustering algorithms led to the identification of distinct DLBCL subtypes reflecting tumor-intrinsic features. The OxPhos-DLBCL subtype is characterized by enhanced mitochondrial oxidative phosphorylation, a major source of potentially toxic reactive oxygen species (ROS). Therefore, we investigated the role of potential mechanisms attenuating ROS toxicity in this DLBCL subtype. We found significantly increased thioredoxin (TXN) mRNA abundance in DLBCLs classified as OxPhos subtype compared to other subtypes. The overall survival (OS) of patients with high TXN mRNA expression was significantly shorter than of those with low TXN mRNA expression, regardless of treatment regimen (R-CHOP and CHOP). TXN overexpression in OxPhos-DLBCLs was also confirmed in a cell line panel at protein level using immunoblotting. To explain transcriptional mechanisms responsible for differential TXN expression in different DLBCLs, we analyzed the TXN promoter and identified two BCL6 binding sites within 1kb upstream of TXN transcription start site. Unlike in other molecular subtypes, BCL6 does not exhibit a repressor activity in OxPhos-DLBCLs (PNAS, 2007; 104: 3207-12). Using luciferase reporter assays and shRNA-mediated gene expression knock-down, we demonstrated that relative differences in TXN abundance between DLBCL subtypes are at least in part caused by the lack of BCL6 transcription repressor activity. We next tested the consequences of TXN depletion in DLBCLs. We found that OxPhos cells with silenced TXN expression were uniformly more sensitive to apoptosis induced by ROS production than control cells. TXN inhibition sensitized all tested cell lines to doxorubicin, the fundamental drug used in DLBCL chemotherapy acting in part as a ROS inducer. In addition to its role in maintaining redox homeostasis, TXN also regulates transcriptional responses to ROS. For example, TXN reduces disulfide bonds between FOXO4 and acetyltransferase p300, which results in reduced FOXO4 acetylation and impaired proapoptotic signaling. For this reason, we assessed whether p300 and TXN are involved in acetylation of FOXO1, a major FOXO member expressed in DLBCLs. In cells with blocked TXN activity, FOXO1 acetylation was significantly higher compared to cells overexpressing wild-type TXN. TXN decreased p300-mediated FOXO1 acetylation, reduced its proapoptotic activity and expression of FOXO1-dependent genes (TRAIL, p27, BIM). We found that FOXO1 and p300 interact in redox and TXN-dependent manner and identified a conserved FOXO1's Cys612 to be responsible for FOXO1-p300 binding. We mutated FOXO1 C612 to Ala and found that when cotransfected with p300, C612A FOXO1 exhibited dramatically suppressed ROS-induced acetylation. Blockade of FOXO1 acetylation resulted in markedly lower expression of FOXO1 target genes, higher cellular proliferation and lower apoptosis. Furthermore, TXN inhibited FOXO1 nuclear translocation in response to oxidative stress in OxPhos-DLBCLs. Finally, silencing FOXO1 in OxPhos cells with knocked-down TXN expression markedly inhibited DLBCL cell line apoptosis in response to oxidative stress, suggesting that FOXO1 is an essential TXN-regulated sensor and effector of ROS toxicity in OxPhos-DLBCL cells. Taken together, these results demonstrate that TXN is overexpressed in a subset of DLBCLs, and high TXN mRNA abundance is related to shorter OS of DLBCL patients. TXN knock-down enhances oxidative stress toxicity in OxPhos cell lines at least in part by facilitating FOXO1 nuclear retention and increasing acetylation of this transcription factor, thus augmenting its proapoptotic activity. Disclosures No relevant conflicts of interest to declare.

Abstract P622: Regulation of Neuroprotective Myeloid Cell Leukemia 1 by Rapamycin and AT2R Agonists in Dopaminergic Neuronal Cell-line

Hypertension ◽  
2015 ◽  
Vol 66 (suppl_1) ◽  
Author(s):  
Jamal Bajwa ◽  
Lakshmi Pulakat
Keyword(s):  
Oxidative Stress ◽  
Cell Death ◽  
Cell Line ◽  
Neuronal Damage ◽  
Neuronal Cells ◽  
Neuronal Cell ◽  
Myeloid Cell ◽  
Serum Starvation ◽  
Cell Leukemia ◽  
Neuronal Cell Line

Myeloid Cell Leukemia I (MCL-1) is a critical protein for neuronal cell survival. MCL-1 is one of the anti-apoptotic proteins in the Bcl2 family. In neurons, MCL-1 regulates the rate of programmed cell death during development and after neuronal damage. It is now well established that without sufficient MCL-1 dopaminergic neuronal cells succumb to cell death under conditions of oxidative stress that result in neurodegenerative diseases such as Parkinsonism. Therefore, identifying drugs that can up-regulate the expression of MCL-1 in neuronal cells is critical for enhancing neuronal resistance to oxidative stress and improving neuronal survival. Aim of this study was to evaluate the effects of treatments with Rapamycin and a novel AT2R peptide agonist NP-6A4 on the MCL-1 expression in SH-SY5Y neuronal cell line. SH-SY5Y cells are a human-derived in vitro model of neuronal function and differentiation, expressing both adrenergic and dopaminergic markers. This cell line is a highly translational model for Parkinson's disease. Cells were maintained in a 1:1 mixture of DMEM and Ham's F-12 with 10% FBS. Cells were subjected to serum starvation and were treated with Rap (10nM), NP-6A4 (300nM) or their combination for 6 hours. MCL-1 protein expression was assessed by immunofluorescence using anti-MCL-1 antibody and a fluorophore-conjugated secondary antibody. Cells were imaged using a confocal microscope and fluorescence was quantified using Leica LAS AF software. It was observed that Rap treatment significantly suppressed MCL-1 expression in SH-SY5Y cells (~40% suppression, p<0.001), whereas Rap+NP-6A4 treatment reversed the Rap-mediated suppression of MCL-1 (p<0.0002). This data indicates that Rapamycin suppresses MCL-1 in dopaminergic neuronal cells and AT2R agonist, NP-6A4 is capable of reversing this effect.

scholarly journals The suitability of glioblastoma cell lines as models for primary glioblastoma cell metabolism

2020 ◽  
Author(s):  
Anya Lara Arthurs ◽  
Damien J Keating ◽  
Simon J Conn
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Metabolic Flux ◽  
Krebs Cycle ◽  
Neural Cells ◽  
Glioblastoma Cell ◽  
Preclinical Research ◽  
Primary Glioblastoma

Abstract Background In contrast to most non-malignant cells, cells comprising Glioblastoma multiforme (GBM), a deadly brain tumour with extremely poor prognosis, preferentially utilise glycolysis over oxidative phosphorylation for metabolism in a phenomenon known as the ‘Warburg effect’. As effective treatments for GBM are severely lacking, research into therapeutics targeting the disease’s highly glycolytic state offer a promising avenue to improve patient survival. These studies often employ GBM cell lines for in vitro studies which translate poorly to the in vivo patient context. Methods The metabolic traits of the seven most commonly used GBM cell lines were assessed using a Seahorse Bioscience Metabolic Flux Analyser and compared to primary GBM cells and primary healthy mixed neural cells from the same patients. Results In support of the glycolytic nature of the patient-derived GBM cell lines, basal mitochondrial rate (p = 0.043) and ATP-linked respiration (p < 0.001) were significantly lower than primary adjacent normal cells from the same patient and reserve capacity (p = 0.037) and Krebs Cycle capacity (p = 0.002) were significantly higher for 12 patients. While no cell line was found to accurately replicate all metabolic attributes of primary GBM cells, specific parameters could be modelled by specific lines. Conclusions U251MG, U373MG and D54 lines are recommended for researching mitochondrial metabolism, and the D645 line for researching ATP-linked respiration. The T98G cell line recapitulated glycolysis-related metabolic parameters of the primary GBM cells and is recommended for research relating to glycolysis. These findings can guide preclinical research into the development of novel therapeutics targeting metabolic pathways in GBM.

scholarly journals TMIGD1 is Expressed at Low Levels in Abdominal Adhesion Tissue and Reduces Oxidative Stress in Peritoneal Mesothelial Cells

2020 ◽  
Author(s):  
Yunhua Wu ◽  
Xuqi Li ◽  
Cong Shen ◽  
Zijun Wang ◽  
Dong Liu ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Cell Adhesion ◽  
Cell Line ◽  
Cell Lines ◽  
Mesothelial Cell ◽  
P38 Inhibitor ◽  
Abdominal Adhesion ◽  
Adhesion Tissue ◽  
Low Levels ◽  
Promote Cell Adhesion

Abstract BackgroundPostoperative abdominal adhesion is one of the most commonly observed complications after abdominal surgery. However, there is no effective treatment for adhesion other than enterolysis. Mesothelial cell repair plays an extremely important role in the process of adhesion formation. Here, we hypothesize that transmembrane and immunoglobulin domain-containing 1 (TMIGD1) is expressed at low levels in abdominal adhesion tissue and can reduce oxidative stress and promote cell adhesion in peritoneal mesothelial cells.Materials and MethodsFirst, we performed gene microarray analysis and used qPCR, western blotting, immunohistochemistry and immunofluorescence to detect the expression of TMIGD1 in rat adhesion tissue and normal peritoneal tissue. Then, we established a TMIGD1-overexpressing HMrSV5 cell line and detected ROS, apoptosis, and the mitochondrial membrane potential by the MTT assay, western blotting, flow cytometry with 2’,7’-dichlorofluorescein diacetate (DCFH-DA) as a probe. Furthermore, we examined p38 phosphorylation in different TMIGD1-expressing cell lines and used a p38 inhibitor to determine whether the antioxidant effect of TMIGD1 is dependent on p38. Finally, we evaluated the adhesion ability of different TMIGD1 cell lines using scratch wound and adhesion assays.ResultsTMIGD1 was expressed at low levels in adhesion tissue and at lower levels in mesothelial cells. TMIGD1 overexpression alleviated H2O2-induced oxidative stress injury in human HMrSV5 cell lines. The phosphorylation level of p38 was higher in the TMIGD1-overexpressing cell line, and we found that the effect of TMIGD1 was inhibited by a p38 inhibitor. In addition, TMIGD1 overexpression inhibited mesothelial cell migration and promoted mesothelial cell adhesion.ConclusionTMIGD1 is expressed at low levels in abdominal adhesion tissue and can reduce H2O2-induced oxidative stress by promoting p38 phosphorylation. In addition, TMIGD1 can promote cell adhesion.# These authors contributed equally to this work.

scholarly journals Endocytosis and Lack of Cytotoxicity of Alkyl-Capped Silicon Quantum Dots Prepared from Porous Silicon

Materials ◽  
2019 ◽  
Vol 12 (10) ◽  
pp. 1702 ◽  
Author(s):  
Wipaporn Phatvej ◽  
Harish K. Datta ◽  
Simon C. Wilkinson ◽  
Elaine Mutch ◽  
Ann K. Daly ◽  
...  
Keyword(s):  
Quantum Dots ◽  
Porous Silicon ◽  
Cell Line ◽  
Cell Lines ◽  
Particle Diameter ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Small Intestinal Mucosa ◽  
High Porosity ◽  
Silicon Quantum Dots ◽  
Atp Production

Freely-dissolved silicon quantum dots were prepared by thermal hydrosilation of 1-undecene at high-porosity porous silicon under reflux in toluene. This reaction produces a suspension of alkyl-capped silicon quantum dots (alkyl SiQDs) with bright orange luminescence, a core Si nanocrystal diameter of about 2.5 nm and a total particle diameter of about 5 nm. Previous work has shown that these particles are rapidly endocytosed by malignant cell lines and have little or no acute toxicity as judged by the standard 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay for viability and the Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay for apoptosis. We have extended this work to the CACO-2 cell line, an established model for the human small intestinal mucosa, and demonstrate that neither acute nor chronic (14 days) toxicity is observed as judged by cell morphology, viability, ATP production, ROS production and DNA damage (single cell gel electrophoresis) at doses of 50–200 μ g mL − 1 . Quantitative assessment of the extent of uptake of alkyl SiQDs by CACO-2, HeLa, HepG2, and Huh7 cell lines by flow cytometry showed a wide variation. The liver cell lines (HepG2 and Huh7) were the most active and HeLa and CACO-2 showed comparable activity. Previous work has reported a cholesterol-sensitivity of the endocytosis (HeLa), which suggests a caveolin-mediated pathway. However, gene expression analysis by quantitative real–time polymerase chain reaction (RT-PCR) indicates very low levels of caveolins 1 and 2 in HepG2 and much higher levels in HeLa. The data suggest that the mechanism of endocytosis of the alkyl SiQDs is cell-line dependent.

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