Cell Journal最新文献

Objective: Diabetes is a metabolic disorder with two common categories of complications: microvascular and macrovascular. Cardiovascular diseases (CVDs), which are linked to inflammation in the blood vessels, are the most important complications of diabetes. G protein-coupled receptors (GPCRs) play a crucial role in initiating these inflammatory responses. The aim of this study is to identify GPCRs that may respond to high blood glucose in monocyte cells and have an effect on regulating vascular inflammation.

Materials and methods: In this in-silico and experimental study, first, RNA sequencing (RNAseq) data from monocytes from the blood of diabetic patients were analyzed to find the GPCR most responsive to high blood sugar. Next, the expression correlation of the candidate GPCR with inflammatory genes was calculated, and expression changes in THP-1 cells under hyperglycemia were evaluated via quantitative polymerase chain reaction (qPCR). Finally, the effect of the candidate GPCR on down-regulation of NF-kB mRNA expression, generation of reactive oxygen species (ROS), and caspase activity was evaluated.

Results: The RNAseq analysis showed that expression of Calcitonin receptor-like (CALCRL), one of the GPCR candidates, increased in the monocytes of patients with diabetes compared to healthy individuals. The qPCR result also indicated that expression of the CALCRL gene increased in THP-1 cells treated with 25 mM D-glucose. Correlation analyses demonstrated that increased expression of CALCRL is associated with inflammatory pathways, cytokine production and secretion, and the cell migration process. Finally, our data indicated that CALCRL downregulation decreased inflammation related to hyperglycemia, including NF-kB mRNA expression, ROS production, and caspase 3/7 activity.

Conclusion: Hyperglycemia increased the expression of CALCRL GPCR in the monocytes of diabetic patients. The CALCRL receptor may play a key role in regulating vascular inflammation in hyperglycemic conditions. Targeting CALCRL may control inflammation in blood cells.

Objective: Aggressive cancer cells exhibit many phenotypic and functional similarities to embryonic stem cells (ESCs), including characteristics such as self-renewal, differentiation flexibility, and specific gene expression signatures. While the ESC microenvironment precisely regulates proliferation to balance self-renewal and differentiation, it remains unclear whether tumor cells can respond to these regulatory signals to suppress their tumorigenic phenotype. Extracellular vesicles (EVs) released by human ESCs (hESCs) are known to exert paracrine effects, making them prime candidates for carrying these regulatory signals. The aim of this study was to determine if hESC-EVs suppress the tumorigenic phenotype of aggressive cancer cells by affecting their proliferative, invasive, and stem cell properties.

Materials and methods: In this experimental study, EVs were isolated from hESC-conditioned medium (hESC-CM) and thoroughly characterized. Aggressive breast and melanoma cancer cell lines were treated with the isolated hESCderived EVs (hESC-EVs). The resulting effects on cell growth (proliferation), invasion, stem cell properties (stemness), and overall tumorigenesis were evaluated in vitro and in vivo. Expression levels of key pluripotency markers were also assessed in the treated cancer cells.

Results: The treatment of the aggressive cancer cell lines with hESC-EVs resulted in a significant reduction in proliferation, invasion, stemness, and tumorigenesis across both in vitro and in vivo models. Mechanistically, exposure to hESC-EVs led to the clear downregulation of pluripotency markers within the recipient cancer cells.

Conclusion: These findings suggest that hESC-EVs transmit powerful regulatory signals capable of inducing the loss of stem cell characteristics in aggressive cancer cells. This mechanism offers a potential pathway for reprogramming highly malignant tumor cells toward a more differentiated and less aggressive phenotype, suggesting a novel strategy for anti-cancer therapy.

Objective: Selecting the most competent sperm is crucial for improving the success of intracytoplasmic sperm injection (ICSI). Chemotaxis-based sperm selection may facilitate the selection of spermatozoa with superior motility and fertilization capability. This study aimed to evaluate sperm selection based on chemotaxis induced by microvesicles (MVs) derived from follicular fluid (FF) and cumulus cells (CCs) using a simple microfluidic device.

Materials and methods: In this experimental study, FF and CCs were collected during ovum pick-up from 10 women undergoing ICSI. MVs from both FF and CCs were isolated using the Exocib kit and characterized for particle size, polydispersity index, zeta potential, morphology, and protein content. To establish a chemoattractant gradient, MVs from FF and CCs, as well as progesterone, were incorporated into agar plates. Nine experimental groups were studied: FF- derived MVs (FF-MV, 5, 25, and 50 μg/ml), CC-derived MV (CC-MV, 5, 25, and 50 μg/ml), FF (50 μg/ml), progesterone (0.03 μM), and a control (agar without chemoattractant). Sperm selection was performed in a microfluidic system using semen samples from 20 normozoospermic men. Sperm concentration, motility, progressive motility, viability, and DNA integrity were analyzed following chemotaxis-based selection.

Results: The isolated FF-and CC-derived MVs exhibited spherical morphology, a negative zeta potential, and were polydispersed. The progesterone-containing plates showed sustained release. No significant differences were observed between the CC-MV and control groups in sperm parameters (P=0.630). Plates supplemented with 50 μg/mL of FF-MV markedly improved sperm concentration (P=0.002), progressive motility (P=0.010), and DNA integrity (P=0.001) compared to the control group.

Conclusion: FF-MVs, particularly at 50 μg/ml, possess chemoattractant properties and could selectively enrich sperm populations with improved functional parameters. These findings highlight the potential of FF-MVs as a novel tool for high-quality sperm selection in the ICSI procedure.

Objective: Neuroinflammation plays a crucial role in neuropathic pain, of which toll-like receptor 4 (TLR4) is a key mediator. Virtual screening identified Drospirenone as a potential TLR4 inhibitor. This study aims to evaluate the in vitro effects of Drospirenone on TLR4 signalling in U87-MG astrocytoma cells under different inflammatory conditions.

Materials and methods: In this experimental study, three treatment approaches with Drospirenone were employed to model various stages of neuroinflammation: co-incubation with lipopolysaccharides (LPS) to assess preventive effects; treatment after inflammation to evaluate its impact on sustained inflammation; and delayed treatment after LPS removal to investigate its role in reducing persistent inflammation. Expression levels of TLR4, myeloid differentiation primary response 88 (MyD88), nuclear factor-κB (NF-κB) p65, and interleukin-1 beta (IL-1β) were assessed by Western blotting, while nitric oxide (NO) secretion was measured by ELISA.

Results: The potential ability of Drospirenone to inhibit early-stage inflammation was shown by significant reductions in TLR4, MyD88, NF-κBp65, IL-1β, and NO. Treatment after inflammation showed that Drospirenone reduced NO secretion but did not significantly affect TLR4 and other inflammatory markers, which could indicate its potential efficacy in controlling sustained inflammation. In the delayed treatment approach, inflammation persisted after LPS removal, and Drospirenone did not return the inflammatory state to baseline.

Conclusion: Drospirenone may exhibit potential as a prophylactic agent in vitro during the early phases of neuroinflammation, though its efficacy appears limited in models of chronic or prolonged inflammation. These preliminary findings require in vivo validation, and future studies could explore possible synergistic effects with other treatments or alternative dosing strategies for neuropathic pain management.

Objective: Human embryonic stem cells (hESCs) have the capacity to differentiate into all cell lineages, making them invaluable for cell-based therapies. However, the ability of individual hESC lines to generate the three embryonic germ layers varies. Selecting a cell line with a high propensity for definitive endoderm (DE) differentiation is crucial for producing pancreatic progenitors, hepatocytes, and other endodermal derivatives. Direct and spontaneous differentiation are two commonly used methods to assess hESC differentiation potential; both are complex and time-consuming. This study evaluates a commercially available differentiation kit as a simple and reproducible screening assay to predict the DE differentiation potential of various hESC lines.

Materials and methods: In this experimental study, three hESC lines obtained from the Royan Institute Stem Cell Bank (Tehran, Iran) were used. Pluripotency was confirmed by evaluating cell morphology, OCT4 protein expression, and alkaline phosphatase staining. Early-stage lineage differentiation was compared between the StemMACS™ Trilineage kit and a previously established DE protocol. Differentiation outcomes were assessed using gene and protein expression analyses, including flow cytometry, quantitative real-time polymerase chain reaction (qRT-PCR), and immunostaining.

Results: The three hESC lines exhibited varying capacities for DE differentiation. Flow cytometry analysis of the surface marker CXCR4 showed differentiation efficiencies of 70% for RH2, compared with 35% and 40% for RH5 and RH6, respectively. The superior efficiency of RH2 was consistent across both differentiation protocols, as evidenced by its strong CXCR4 expression using the in-house method.

Conclusion: hESC lines exhibit variable capacities for differentiation. Commercial kit-based screening provides a simple and reliable method to identify suitable hESC lines for generating endoderm-derived cells.

Objective: Breast cancer remains one of the leading causes of mortality among women worldwide. Approximately 80% of patients receive radiotherapy (RT) as part of their treatment. Numerous efforts have been made to distinguish radiosensitive from radioresistant breast tumors. Long non-coding RNAs (lncRNAs) have increasingly been recognized as crucial mediators of cellular responses to radiation among the wide range of biomarkers studied. This study explored the radiation-induced expression changes of three apoptosis-associated lncRNAs-MEG3, MALAT1, and ANRIL-in breast cancer.

Materials and methods: In this experimental study, two established human breast cancer cell lines, MCF-7 and MDAMB- 231, were cultured in Dulbecco's Modified Eagle Medium (DMEM) and irradiated with X-rays at doses ranging from 0 to 8 Gy. The expression levels of the lncRNAs ANRIL, MALAT1 and MEG3 were quantified by real-time polymerase chain reaction (PCR). Cell viability in irradiated MCF-7 and MDA-MB-231 cells was evaluated using the MTT colorimetric assay.

Results: Following radiation exposure, the expression levels of ANRIL and MEG3 were significantly decreased (P=0.004) in both MCF-7 and MDA-MB-231 cell lines, whereas MALAT1 expression was significantly increased (P=0.006) only in MDA-MB-231 cells. In addition, cell viability in both MCF-7 and MDA-MB-231 lines declined after irradiation.

Conclusion: Our findings indicate that radiation exposure increases MALAT1 expression while decreasing ANRIL and MEG3 levels, associated with reduced cell viability .These results suggest a potential role for these lncRNAs in radiation-induced cell death.

Inosine-5'-monophosphate dehydrogenase (IMPDH) serves as a key regulatory enzyme, catalyzing a crucial step in the de novo synthesis of guanine nucleotides. This enzyme plays a pivotal role in various cellular functions, including proliferation, differentiation, homeostasis, and metabolism. IMPDH has the unique ability to form protein polymer fibers known as IMPDH cytoophidia. These natural biological structures are integral to purine metabolism and play a significant role in regulating changes within the cellular environment. Utilizing advanced techniques such as immunofluorescence microscopy and Western blot analysis, we have demonstrated that ribavirin (RBV) exhibits remarkable efficacy in inducing time-dependent cytoophidia formation when compared to other IMPDH inhibitors. Our observations reveal that small filaments progressively aggregate into larger secondary structures, reaching their peak formation at 12 hours. Subsequently, the structures begin to decrease by 24 hours, and ringlike formations dominate the cellular landscape at a remarkable 62% incidence. These findings suggest a complex and dynamic process of cytoophidia maturation in response to prolonged and time-dependent IMPDH inhibition. This process underscores the sequential emergence and development of secondary structures that surpass the conventional ring-rod configuration. Our study provides valuable insights into the mechanisms underlying cytoophidia formation and their potential implications for cellular regulation and metabolism.

Objective: Hepatocellular carcinoma (HCC), a prevalent and aggressive malignancy, is one of the most common malignancies worldwide. Various studies show that ubiquitin D (UBD) is overexpresses in different cancer types and may serve as a potential prognostic factor. Although catenin alpha 3 (CTNNA3) is a tumour suppressor in HCC, the relationship between UBD and CTNNA3 in HCC remains unclear. This study aims to explore the role of UBD in HCC and its relationship with CTNNA3 in HCC cells.

Materials and methods: In this experimental study, UBD expression in clinical samples was analysed by reverse transcription quantitative polymerase chain reaction (RT-qPCR) and immunohistochemistry. Protein levels of epithelialmesenchymal transition (EMT) markers were evaluated by Western blot. Cellular behaviours that included proliferation, colony formation, migration, and invasion were assessed using the CCK-8, colony formation, EdU, and transwell assays. UBD-mediated ubiquitination of CTNNA3 was examined by in vitro ubiquitination assays.

Results: There was a significant elevation in UBD expression in the HCC patients, which correlated with poor prognosis. Knockdown of UBD suppressed the proliferation, colony formation, and EMT of the HCC cells. UBD reduced CTNNA3 expression in HCC cells by promoting the ubiquitination and degradation of CTNNA3. Decreased CTNNA3 expression in HCC patients was associated with poor overall survival. Silencing of CTNNA3 repressed HCC proliferation, migration, invasion, and EMT. CTNNA3 deficiency counteracted the inhibitory effects of UBD knockdown on the malignant behaviour of the HCC cells.

Conclusion: UBD plays an oncogenic role in HCC by encouraging proliferation and EMT through promoting CTNNA3 degradation. These findings suggest that targeting the UBD/CTNNA3 axis could be a potential therapeutic strategy for HCC management.

Depression is among the most widespread mental health conditions worldwide, with a significant number of patients showing inadequate responses to pharmacological treatments. Despite the development of numerous pharmacological therapies over the past six decades, a substantial proportion of patients with major depressive disorder (MDD) remain unresponsive to conventional therapies, prompting the exploration of innovative therapeutic strategies. This review critically examines the potential of stem cell-based therapies as a novel alternative to conventional antidepressant treatments, with a particular focus on individuals diagnosed with MDD. This study explores the molecular pathways that mediate neuroinflammatory responses in MDD, emphasizing the contributions of key pro-inflammatory cytokines to impaired neuroplasticity, mitochondrial dysfunction, and heightened oxidative stress. In addition, this study discusses the immunomodulatory properties of mesenchymal stromal cells (MSCs), highlighting their capacity to enhance antiinflammatory signaling, promote neurogenesis, and restore neurotransmitter homeostasis. Moreover, this review underscores the neuroprotective role of MSCs in mitigating oxidative stress-induced neuronal injury, with particular emphasis on their ability to secrete neurotrophic factors such as brain-derived neurotrophic factor (BDNF), which support neuronal integrity and survival.

Objective: Acute myeloid leukemia (AML) is a heterogeneous malignancy driven by disruptions in the bone marrow microenvironment (BME). Extracellular vesicles (EVs) are crucial communicators and effectors in the BME, facilitating interactions between leukemic cells and stromal components to promote leukemogenesis. Elucidating these EVmediated processes is essential for developing novel therapeutic strategies. This study sought to clarify the effects of EVs derived from newly diagnosed non-M3 AML patients on bone marrow mesenchymal stromal cells (BMSCs), focusing on proliferation, survival, apoptosis, and expression of CTNNB1, TGF-β, and VEGF genes, which are pivotal in leukemia progression.

Materials and methods: In this experimental study, AML-derived EVs were isolated from 30 newly diagnosed non-M3 AML patients and characterized by dynamic light scattering (DLS), transmission electron microscopy (TEM), Bradford assay, and flow cytometry. BMSCs were isolated from healthy BM aspirates and were co-cultured with EVs at concentrations of 10, 40, and 60 μg/ml for 24, 48, and 72 hours. Outcomes were assessed using quantitative reverse transcription polymerase chain reaction (RT-PCR), (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliuom bromide (MTT) assay, flow cytometry [for apoptosis, reactive oxygen species (ROS), and Ki67], and Western blot analyses.

Results: AML-derived EVs at 40 μg/ml significantly suppressed apoptosis, enhanced cell survival, and upregulated CTNNB1, TGF-β, and VEGF gene expression in BMSCs via the Wnt/β-catenin signaling pathway, as confirmed by increased β-catenin protein levels. However, a 60 μg/ml dose increased apoptosis and reduced gene expression.

Conclusion: These findings can suggest that AML-derived EVs modulate the BME by promoting BMSC survival and upregulating pro-leukemic genes at an optimal dose, offering a potential therapeutic target for AML treatment.