PMCID stringclasses 30 values | sentence stringlengths 1 1.04k | entities listlengths 0 22 |
|---|---|---|
PMC12842104 | At least three separate experiments, each containing three replicates, were performed. | [] |
PMC12842104 | HMECs showed reduction in cell viability starting at 125 µg/mL concentration using E. bicolor xylene extract. | [] |
PMC12842104 | Therefore, the rest of the experiments were set up to use 62.5 µg/mL concentration to determine the antiproliferative mechanisms. | [] |
PMC12842104 | The 50% inhibitory concentration (IC50) was estimated using the GraphPad Prism 9.4 software as described before . | [] |
PMC12842104 | The IncuCyte live cell imaging system (Sartorius, Ann Arbor, MI, USA) was used to observe and capture photos at 4 h intervals of morphological changes of treated cells for several days. | [] |
PMC12842104 | Cells were seeded into a 96-well plate (10,000 cells/well) and exposed to 62.5 µg/mL of E. bicolor xylene extract, before being placed into the IncuCyte . | [] |
PMC12842104 | Cell apoptosis was assessed with the Click-iT™ Plus TUNEL assay kit (ThermoFisher Scientific, Waltham, MA, USA) as reported previously . | [] |
PMC12842104 | Cells were treated with E. bicolor xylene extract at a concentration of 62.5 µg/mL for 24 h before proceeding with the TUNEL assays. | [] |
PMC12842104 | Cells were pretreated with 10 µM of the TRPV1 antagonist capsazepine (Abcam, USA) for 30 min as previously described before being treated with E. bicolor xylene extract (62.5 µg/mL) and cell viability measured using MTS assays. | [] |
PMC12842104 | Three independent experiments were conducted. | [] |
PMC12842104 | Cells were pretreated with 1 µM of the calcium chelator BAPTA-AM (Abcam, San Francisco, CA, USA), as previously described , and then treated with E. bicolor xylene extract (62.5 µg/mL) before MTS assays for cell viability were performed. | [] |
PMC12842104 | Three independent experiments were conducted. | [] |
PMC12842104 | The cytoplasmic calcium dynamics as the result of TRPV1 activation were observed using Fura2-AM staining. | [] |
PMC12842104 | T47D and MDA-MB-231 cells were seeded into a 6-well plate and cultured overnight. | [
{
"end": 4,
"label": "CellLine",
"start": 0,
"text": "T47D"
},
{
"end": 19,
"label": "CellLine",
"start": 9,
"text": "MDA-MB-231"
}
] |
PMC12842104 | After 24 h, cells were loaded with 5 μM of Fura2-AM for 30 min . | [] |
PMC12842104 | Following E. bicolor xylene treatment, the cytosolic Ca signal was monitored continuously for 20 min using a LionHeart FX microscope (Agilent, Santa Clara, CA, USA). | [] |
PMC12842104 | The intracellular ROS levels were determined using cell-permeant reagent 2’,7’-dichlorofluorescin diacetate (DCFDA) according to the manufacturer’s instructions (Abcam). | [] |
PMC12842104 | Tert-butyl hydroperoxide (TBHP; 250 μM) was used as positive control. | [] |
PMC12842104 | The fluorescence intensity values at each time point were calculated as the ratio of the value at a specific t-time point to the value at point zero time (t-time point/t0); t0 = first measurement. | [] |
PMC12842104 | Briefly, cells were plated in FBS-supplemented medium without phenol red onto 96-well plates. | [] |
PMC12842104 | After 24 h, the cells were washed once with 1× buffer provided in the kit, then the cells were incubated with 10 μM of DCFDA for 30 min at 37 °C, protected from light. | [] |
PMC12842104 | Following incubation, the wells were washed with PBS, and treated with E. bicolor xylene extract. | [] |
PMC12842104 | ROS production was determined immediately by measuring the formation of fluorescent dichlorofluorescein (DCF), using a Synergy microplate reader (Agilent, Winooski, VT, USA), at 485 nm excitation and 535 nm emission. | [] |
PMC12842104 | Measurements were taken every 60 min for six hours. | [] |
PMC12842104 | Three independent experiments were conducted. | [] |
PMC12842104 | Cells were pretreated with NAC (5 mM) for 1 h followed by treatment with or without E. bicolor xylene extract for another 72 h, and cell viability was measured using MTS assay. | [] |
PMC12842104 | Three independent experiments were conducted. | [] |
PMC12842104 | ER-targeted low-affinity GCaMP6-210 variant, a fluorescent reporter for ER calcium signaling, was used to observe endoplasmic reticulum (ER) Ca dynamics. | [] |
PMC12842104 | T47D cells were seeded into a 24-well plate at 10.4 × 10 cells in 500 µL phenol red free growth medium. | [
{
"end": 4,
"label": "CellLine",
"start": 0,
"text": "T47D"
}
] |
PMC12842104 | After 24 h, cells were transfected with the GCaMP6-210 variant using Invitrogen™ Lipofectamine™ 3000 Transfection Reagent (Thermofisher Scientific, Waltham, MA, USA) according to the manufacturer’s instructions. | [] |
PMC12842104 | Twelve hours after incubation, immediately after E. bicolor xylene extract treatment (62.5 µg/mL), calcium influx was observed with the LionHeart FX microscope. | [] |
PMC12842104 | The mitochondrial calcium dynamics were observed using Rhod2-AM (Abcam, USA) as previously described . | [] |
PMC12842104 | Cells simultaneously loaded with 5 μM of Rhod2-AM, and 10 μM of MitoTracker Green (ThermoFisher Scientific, Waltham, MA, USA) were treated with E. bicolor xylene extracts (62.5 µg/mL) and fluorescence signals were monitored for 10 min with the LionHeart FX microscope. | [] |
PMC12842104 | Cells treated with E. bicolor xylene extract (62.5 µg/mL) for 24 h and incubated at 37 °C for 24 h. Detection of caspase 3 activation with the CellEvent™ Caspase-3/7 Green ReadyProbes™ reagent (ThermoFisher Scientific, Waltham, MA, USA) was performed as described before . | [] |
PMC12842104 | As previously reported , Western blotting was carried out with a few changes. | [] |
PMC12842104 | Proapoptotic and anti-apoptotic proteins were examined using total cellular protein. | [] |
PMC12842104 | Preparation of cell lysates, estimation of total protein concentrations, SDS-PAGE, transfers onto polyvinylidene difluoride (PVDF) membranes (BioRad, Hercules, CA, USA), and probing with antibodies follow the procedure described before . | [] |
PMC12842104 | Antibodies used were as follows: anti-beta actin, anti-caspase 9, anti-caspase 8, anti-CHOP, anti-ATF4, anti-FAS, anti-PERK (mouse monoclonal antibody conjugated with HRP; 1:1000, v/v) (Santa Cruz Biotechnology, Dallas, TX, USA), anti-AKT, anti-pAKT (mouse monoclonal antibody conjugated with Alexa 488; 1:1000, v/v) (Santa Cruz Biotechnology, USA). | [] |
PMC12842104 | Membranes were washed with Tris-buffered saline containing 0.1% (v/v) Tween 20 (TBST), incubated with enhanced chemiluminescence substrate solution (BioRad, USA), according to the manufacturer’s instructions, and visualized with a ChemiDoc system. | [] |
PMC12842104 | For anti-Bcl-2, anti-BAX (rabbit monoclonal antibody; 1:1000, v/v) (Abcam, USA), anti-pPERK, anti-PI3K, anti-XBP1s (rabbit monoclonal antibody; 1:1000, v/v) (Cell Signaling Technology, Danvers, MA, USA), following the overnight incubation, the membranes were incubated with the secondary antibody (Goat Anti-Rabbit IgG H&L, Alexa Fluor 488; 1:500 v/v) (Thermofisher scientific, Waltham, MA, USA) for five minutes each after being rinsed three times in Tris-buffered saline containing 0.1% (v/v) Tween 20 (TBST) for 1 h. Means and standard errors of at least three experiments were calculated. | [] |
PMC12842104 | One-way ANOVA was performed, followed by Tukey’s post hoc test to determine significant differences among the means for the antiproliferative (MTS) assays and fold expression analysis of Western blot (to compare DMSO control and E. bicolor xylene extract treatment) using GraphPad Prism 9.4. | [] |
PMC12842104 | A p-value < 0.05 was considered statistically significant. | [] |
PMC12842104 | Unpaired Welch’s t-test, to compare treated to DMSO control set, was used for TUNEL assays. | [] |
PMC12842104 | A two-way ANOVA was performed, followed by the Dunnett test to determine significant differences among the means of Western blot fold expression (to compare DMSO control, E. bicolor xylene extract, and capsaicin treatment). | [] |
PMC12842104 | ImageJ 1.5p software, (https://imagej.net/ij/download.html, accessed on 11 November 2023) was used to determine fold protein expression on Western blots. | [] |
PMC12842104 | Our study presents for the first time the ROS-mediated antiproliferative activity of E. bicolor xylene extract in T47D cells and TRPV1-dependent antiproliferative activity in MDA-MB-231 cells and their mechanisms of action. | [
{
"end": 118,
"label": "CellLine",
"start": 114,
"text": "T47D"
},
{
"end": 185,
"label": "CellLine",
"start": 175,
"text": "MDA-MB-231"
}
] |
PMC12842104 | E. bicolor xylene extract generates high ROS levels in T47D cells and triggers several apoptotic pathways. | [
{
"end": 59,
"label": "CellLine",
"start": 55,
"text": "T47D"
}
] |
PMC12842104 | In contrast, E. bicolor xylene extract activates TRPV1 and induces mitochondrial and endoplasmic reticulum stress-mediated apoptotic pathways in MDA-MB-231 cells. | [
{
"end": 155,
"label": "CellLine",
"start": 145,
"text": "MDA-MB-231"
}
] |
PMC12842104 | In addition, E. bicolor xylene extract downregulates the PI3K/AKT signaling pathway in both T47D and MDA-MB-231 cells. | [
{
"end": 96,
"label": "CellLine",
"start": 92,
"text": "T47D"
},
{
"end": 111,
"label": "CellLine",
"start": 101,
"text": "MDA-MB-231"
}
] |
PMC12842104 | Our findings suggest that E. bicolor biochemicals could be used to design cancer cell type-specific therapeutics. | [] |
PMC12514566 | Epstein–Barr virus-associated gastric carcinoma (EBVaGC), a distinct subtype of gastric cancer, accounts for approximately 10 % of all gastric cancer cases. | [] |
PMC12514566 | 2-deoxyglucose (2-DG), a glycolysis inhibitor, has emerged as a crucial tool in cancer therapy. | [] |
PMC12514566 | However, the differential effects of 2-DG on EBVaGC and EBV-negative gastric carcinoma (EBVnGC) are not yet fully understood. | [] |
PMC12514566 | In this study, we demonstrated that 2-DG inhibited the proliferation of both AGS and AGS-EBV cells, with AGS-EBV cells exhibiting greater sensitivity, particularly under hypoxic conditions. | [
{
"end": 80,
"label": "CellLine",
"start": 77,
"text": "AGS"
},
{
"end": 92,
"label": "CellLine",
"start": 85,
"text": "AGS-EBV"
},
{
"end": 112,
"label": "CellLine",
"start": 105,
"text": "AGS-EBV"
}
] |
PMC12514566 | Furthermore, EBV infection was found to upregulate glycolytic gene expression in AGS-EBV cells, particularly under hypoxic conditions, through HIF-1α-dependent mechanisms. | [
{
"end": 88,
"label": "CellLine",
"start": 81,
"text": "AGS-EBV"
}
] |
PMC12514566 | Notably, 2-DG also inhibited EBV lytic reactivation in AGS-EBV cells under hypoxic conditions. | [
{
"end": 62,
"label": "CellLine",
"start": 55,
"text": "AGS-EBV"
}
] |
PMC12514566 | These findings provide valuable insights into the molecular mechanisms of EBV-mediated metabolic reprogramming and highlight the potential of 2-DG as a therapeutic agent for EBVaGC.Epstein–Barr virus (EBV) infects over 90 % of the global population and is a well-established contributor to various human malignancies. | [] |
PMC12514566 | Among EBV-related cancers, EBV-associated gastric carcinoma (EBVaGC) is the most prevalent subtype, with an estimated 75,000 to 90,000 new cases diagnosed worldwide each year. | [] |
PMC12514566 | Representing approximately 5–10 % of all gastric cancers, EBVaGC is recognized as an independent subtype with distinct clinical, pathological, and molecular characteristics.2, 3, 4 The gold standard for its diagnosis is the detection of EBV-encoded small RNAs (EBERs) through in situ hybridization (ISH). | [] |
PMC12514566 | EBERs-ISH results consistently show that both cancer cells and adjacent dysplastic epithelial cells in EBVaGC are EBERs-positive, while surrounding normal gastric mucosal epithelial cells and lymphocytes are EBERs-negative, strongly suggesting that EBV infection precedes malignant transformation. | [] |
PMC12514566 | In EBVaGC, EBV typically maintains a non-classical latent type I infection stage, primarily expressing viral products such as EBNA1, EBERs, ebv-miR-BARTs, and ebv-circRNAs, with LMP2A detectable in approximately 50 % of cases.7, 8, 9 These viral factors play crucial roles in maintaining latency and contributing to oncogenesis. | [] |
PMC12514566 | Metabolic reprogramming is a well-established hallmark of cancer, where cells often adopt glycolysis as their primary energy source, even in the presence of sufficient oxygen, a phenomenon known as the "Warburg effect". | [] |
PMC12514566 | While glycolysis is less efficient at producing ATP than oxidative phosphorylation, its enhanced glycolytic rate in tumor cells provides key metabolites necessary for rapid cell growth and division. | [] |
PMC12514566 | This metabolic shift is particularly evident in solid tumors, including gastric cancer, where hypoxic microenvironments frequently develop due to uncontrolled tumor growth outcompeting vascular oxygen supply and aberrant neovascularization. | [] |
PMC12514566 | The cellular adaptation to hypoxia is largely orchestrated by hypoxia-inducible factors (HIFs), particularly HIF-1α and HIF-2α. | [] |
PMC12514566 | In gastric cancer, studies utilizing gas chromatography-mass spectrometry (GC/MS) have identified significant increases in key glycolytic metabolites, such as fumarate and α-ketoglutarate, in tumor tissues compared to healthy controls. | [] |
PMC12514566 | Other studies have reported elevated serum levels of metabolites like 3-hydroxypropionic acid and pyruvate in gastric cancer patients, further emphasizing the critical role of glycolysis in gastric cancer progression. | [] |
PMC12514566 | Given the heavy reliance of cancer cells on glycolysis, targeting metabolic pathways, particularly glycolysis, has emerged as an attractive therapeutic strategy. | [] |
PMC12514566 | However, the metabolic characteristics of EBVaGC in comparison to EBVnGC, particularly with respect to glycolysis, remain poorly understood. | [
{
"end": 48,
"label": "CellLine",
"start": 42,
"text": "EBVaGC"
},
{
"end": 72,
"label": "CellLine",
"start": 66,
"text": "EBVnGC"
}
] |
PMC12514566 | While some studies have highlighted glycolytic alterations in gastric cancer, it is unclear whether EBVaGC exhibits distinct metabolic features or differences in glycolysis and its response to glycolysis inhibitors. | [] |
PMC12514566 | Investigating these differences could provide valuable insights into precision metabolic therapies tailored to EBVaGC, offering potential new strategies for the treatment of this challenging subtype of gastric cancer. | [] |
PMC12514566 | In this study, we explored the effects of 2-DG on AGS and AGS-EBV cells, discovering that AGS-EBV cells exhibited greater sensitivity to 2-DG, particularly under hypoxic conditions. | [
{
"end": 53,
"label": "CellLine",
"start": 50,
"text": "AGS"
},
{
"end": 65,
"label": "CellLine",
"start": 58,
"text": "AGS-EBV"
},
{
"end": 97,
"label": "CellLine",
"start": 90,
"text": "AGS-EBV"
}
] |
PMC12514566 | EBV infection enhanced glycolytic gene expression in AGS-EBV cells through a HIF-1α-dependent mechanism. | [
{
"end": 60,
"label": "CellLine",
"start": 53,
"text": "AGS-EBV"
}
] |
PMC12514566 | Furthermore, treatment with 2-DG significantly reduced EBV lytic reactivation in EBVaGC cells under hypoxic conditions. | [
{
"end": 87,
"label": "CellLine",
"start": 81,
"text": "EBVaGC"
}
] |
PMC12514566 | Our findings provide new insights into EBV-mediated metabolic reprogramming and underscore the potential of 2-DG as a promising therapeutic approach for EBVaGC. | [] |
PMC12514566 | The gastric carcinoma cell lines AGS and SNU719 are cultured in RPMI-1640 medium with 10 % FBS. | [
{
"end": 36,
"label": "CellLine",
"start": 33,
"text": "AGS"
},
{
"end": 47,
"label": "CellLine",
"start": 41,
"text": "SNU719"
}
] |
PMC12514566 | Akata-EBV-GFP is an Akata Burkitt lymphoma cell line that carries a GFP-tagged Akata bacterial artificial chromosome (BAC) and is cultured in RPMI-1640 medium with 10 % fetal bovine serum (FBS) (Gibco, Carlsbad, CA, USA). | [
{
"end": 25,
"label": "CellLine",
"start": 20,
"text": "Akata"
},
{
"end": 47,
"label": "CellLine",
"start": 26,
"text": "Burkitt lymphoma cell"
}
] |
PMC12514566 | The GFP fluorescein gene, introduced via recombinant EBV, is integrated into the open reading frame of the lytic gene BXLF1. | [] |
PMC12514566 | For EBV production, Akata-EBV-GFP cells were treated with 0.5 % (v/v) goat anti-human IgG for 8 h to induce the transition of EBV from the latent phase to the lytic cycle. | [
{
"end": 33,
"label": "CellLine",
"start": 20,
"text": "Akata-EBV-GFP"
}
] |
PMC12514566 | Afterward, the medium was replaced with fresh medium containing 5 % FBS, and the cells were co-cultured with AGS cells that had been pre-treated with basic medium for 24 h. The culture flask was gently shaken every 4 h to ensure adequate contact between the Akata-EBV-GFP cells and AGS cells. | [
{
"end": 112,
"label": "CellLine",
"start": 109,
"text": "AGS"
},
{
"end": 271,
"label": "CellLine",
"start": 258,
"text": "Akata-EBV-GFP"
},
{
"end": 285,
"label": "CellLine",
"start": 282,
"text": "AGS"
}
] |
PMC12514566 | The medium was partially changed every 24 h, and finally, the Akata cells were thoroughly washed. | [] |
PMC12514566 | The cells were then maintained in culture medium containing 700 μg/mL G418 (Sangon Biotech, Chengdu, China). | [] |
PMC12514566 | Briefly, 1 × 10^5 indicated cells were seeded in a 6-well plate and treated with either 0 or 5 mM 2-DG under normoxic (21 % O2) or hypoxic (1 % O2) conditions for 48 h. Following the treatment, cells were collected and stained with 0.4 % Trypan Blue (1:9 ratio) for 5 min. | [] |
PMC12514566 | Viable cells were then counted using a hemocytometer. | [] |
PMC12514566 | For RNA interference experiments, siRNAs targeting HIF1-α and HIF2-α were synthesized by RiboBio (Guangzhou, China). | [] |
PMC12514566 | Transfections were performed using Lipo3000 (Invitrogen, Life Technologies, Carlsbad, CA, USA) according to the manufacturer's instructions. | [] |
PMC12514566 | Here is the sequence: siHIF-1α: CCAGCAACTTGAGGAAGTA; siHIF-2α: GGCCCTCATTTGAGTCCTA. | [] |
PMC12514566 | Total RNA was isolated from the cells according to the AG RNAex Pro Reagent manual (AG, Changsha, China). | [] |
PMC12514566 | The extracted RNA was then reverse transcribed using the Evo M-MLV RT Kit with gDNA Clean (AG), following the manufacturer's protocol. | [] |
PMC12514566 | The RT-qPCR was carried out on the LightCycler 480 (Roche, Basel, Switzerland) using the SYBR Green Premix Pro Taq HS qPCR Kit (AG) in accordance with the manufacturer's guidelines. | [] |
PMC12514566 | Each sample was analyzed in three biological replicates, with each biological replicate including three technical replicates for robustness and accuracy. | [] |
PMC12514566 | The RT-qPCR data were evaluated using the relative quantification method (2), with ACTB as the internal reference for normalization. | [] |
PMC12514566 | Primer sequences: ACTB-F: CACCATTGGCAATGAGCGGTTC. | [] |
PMC12514566 | ACTB-R: AGGTCTTTGCGGATGTCCACGT. | [] |
PMC12514566 | HIF1α-F: TATGAGCCAGAAGAACTTTTAGGC. | [] |
PMC12514566 | HIF1α-R: CACCTCTTTTGGCAAGCATCCTG. | [] |
PMC12514566 | GLUT1-F: TTGCAGGCTTCTCCAACTGGAC. | [] |
PMC12514566 | GLUT1-R: CAGAACCAGGAGCACAGTGAAG. | [] |
PMC12514566 | HK2-F: GAGTTTGACCTGGATGTGGTTGC. | [] |
PMC12514566 | HK2-R: CCTCCATGTAGCAGGCATTGCT. | [] |
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