Cell Journal最新文献

The retinal pigment epithelium (RPE) cells are a single layer of cells with specific functions in vision. The Mertk gene, encoding a receptor tyrosine kinase, is critical for the phagocytic function of retinal RPE cells. Mutations in Mertk disrupt RPE function and contribute to retinal degeneration. This study aims to characterize the Mertk mutation in Royal College of Surgeons (RCS) rats and develop a polymerase chain reaction (PCR)-based method for genotyping these mutations to improve colony management. DNA was extracted from mutant Mertk^-/- and wild-type rats, followed by PCR amplification using primers flanking the deletion region. Sequencing of the PCR products was performed to identify the precise nature of the mutation. A PCR-based genotyping method was then developed to distinguish between homozygous and heterozygous mutants. Sequencing revealed a 1850 bp deletion in the Mertk gene, resulting in a truncated protein that potentially impairs RPE phagocytosis. The newly developed PCR method successfully differentiated between homozygous and heterozygous mutant rats. This genotyping technique proved to be efficient and reliable, facilitating the management of rat colonies for research purposes. This study provides a detailed molecular characterization of the Mertk mutation in RCS rats, enhancing our understanding of Mertk-related retinal degenerative diseases. The development of a robust PCR-based genotyping method enables efficient differentiation of rat genotypes, aiding in the creation and maintenance of rat models for future research. These findings underscore the importance of molecular characterization in advancing our understanding of genetic models and improving research meth odologies.

Objective: The study aims to investigate the regulatory mechanism of the Fndc5 gene in subcutaneous adipose tissue of diabetic mice, with a focus on the role of miR-129-5p in the pathogenesis of type 2 diabetes mellitus (T2DM). Specifically, it examines how the dysregulation of miR-129-5p affects Fndc5 expression and contributes to diabetesrelated metabolic changes. By exploring these molecular pathways, the research seeks to enhance our understanding of T2DM and identify potential therapeutic targets for its complications.

Materials and methods: In this experimental study, a total of 12 C57BL/6 male mice (6 weeks old) were divided into control and 60% high-fat enriched advanced glycation end products (60% HF-AGE) groups (n=6 per group). Bioinformatics analysis involved mining altered miRNAs in type 2 diabetes and predicting miRNA interactions with Fndc5 mRNA. RNA and proteins extracted from adipose tissue analyzed by using quantitative real-time polymerase chain reaction (PCR) and immunoblotting, respectively. The dual luciferase reporter assay investigated direct interaction between miR-129-5p and the Fndc5 gene using HEK293T cells transfected with relevant vectors.

Results: In mice receiving a 60% HF-AGE diet, significant increases in energy intake, body weight, insulin levels, and fasting blood glucose (FBS) were observed. Furthermore, miR-129-5p was identified as a potential regulator of the Fndc5 gene, displaying elevated expression in diabetic adipose tissue. Plasmid construction confirmed the binding site of miR-129-5p on the Fndc5 3'UTR, while dual luciferase assays validated its direct targeting of the Fndc5 transcript. This interaction corresponded with a reduced expression of Fndc5, highlighting its potential role in diabetes-related metabolic dysregulation.

Conclusion: The upregulation of mir-129-5p in these tissues and the subsequent decrease in the expression of the Fndc5 gene may play a role in developing pathological conditions in this tissue. These findings highlight potential mechanisms linking diet-induced diabetes with Fndc5 regulation through miR-129-5p.

Chronic cervicitis is a clinical syndrome characterized by persistent inflammation of the cervical epithelium. Women with chronic cervicitis often experience symptoms such as vaginal discharge, dyspareunia, post-coital bleeding, and fatigue. The treatment of chronic cervicitis is controversial, particularly in cases where no specific pathogenic agent be identified. Natural wound dressings with human amniotic membranes have been successfully used in tissue healing procedures. Thus, this approach may be a potential therapeutic option for patients suffering from chronic cervicitis. This case series reports on five women diagnosed with chronic cervicitis. These patients had negative Pap smears, human papillomavirus (HPV) typing tests, and colposcopic biopsies, indicating no infections or malignancies. Despite receiving oral antibiotics, their symptoms did not improve, leading to the decision to treat them with amniotic membrane. The cervix was assessed using a speculum, and amniotic membranes manufactured by Sinacell Company were inserted into the affected cervical areas. The patients were instructed to avoid sexual intercourse for one week and continue using oral antibiotics. At the follow-up, all patients significantly improved without any adverse events. Additionally, colposcopic examinations revealed that most patients showed improvement in cervical inflammation and bleeding. The amniotic membrane can assist women with chronic cervicitis by alleviating symptoms and enhancing the healing of cervical ulcers without complications.

Objective: Acrosome reaction is a receptor and Ca²⁺-dependent process. N-Methyl-D- aspartate (NMDA) glutamate receptors are expressed in the human sperm and are involved in the Ca²⁺ influx. Therefore, the present study aimed to investigate whether these receptors could be concerned with the induction of acrosome reaction in the human sperm.

Materials and methods: In this experimental study, human spermatozoa were divided into five groups: i. Spermatozoa at 0 hour, ii. Control group, iii. Spermatozoa treated with NMDA glutamate receptor agonist (L-glutamate), iv. Spermatozoa treated with NMDA glutamate receptor antagonist and agonist (MK-801 and L-glutamate respectively), and v. Spermatozoa treated with non-NMDA glutamate receptor agonist (Kainic acid). Spermatozoa from groups of 2 to 5 were incubated in a Co₂ incubator for 60 minutes. To study the acrosome reaction in the different groups' the sperm samples were stained with Pisum sativum. In addition, cytosolic Ca²⁺ levels were assessed in the experimental groups. One-way ANOVA followed by Tukey's test was used to determine the statistical significance of the data. A P<0.05 was considered significant.

Results: In the present study, applying L-glutamate significantly (P<0.001) induced an acrosome reaction compared to the control group. In the MK-801+ L-glutamate group, MK-801 could significantly (P=0.010) inhibit the induction of acrosome reaction compared to the L-glutamate group. The application of kainic acid did not affect the induction of acrosome reaction compared to the control group. In the L-glutamate group, cytosolic Ca²⁺ levels were significantly (P=0.044) increased when compared with the control group. The application of MK-801+ L-glutamate could significantly (P=0.047) reverse the effect of L-glutamate on cytosolic Ca²⁺ levels compared to the L-glutamate group.

Conclusion: The results of the present study showed that NMDA glutamate receptors play an essential role not only in the increase of cytosolic Ca²⁺ levels but also in the induction of the acrosome reaction in human sperm.

Objective: For years, it has been acknowledged that cancer cells contribute to aberrant DNA methylation, an epigenetic alteration. DNA methyltransferase 1 (DNMT1) is a large multidomain protein critical DNMT in cells. Due to its significant function in epigenetic control, DNMT1 is a viable candidate gene for cancer susceptibility. The relationships between DNMT1 polymorphisms and cancer risk have been investigated; however, the outcomes are inconsistent. This metaanalysis aims to clarify the relationships between DNMT1 polymorphisms and cancer susceptibility.

Materials and methods: PubMed, Web of Science, and Scopus databases were systematically searched using specific search terms to identify potentially eligible papers published before January 2025. Fixed-effects or randomeffects models were employed to calculate odds ratios (OR) and 95% confidence intervals (CI). The I² statistic and Egger's test were utilised to evaluate inter-study heterogeneity and assess the presence of publication bias among the included studies. All statistical analyses were conducted using MetaGenyo software.

Results: A total of 776 articles were retrieved from PubMed, Scopus, and Web of Science databases. After full-text evaluation and applying the literature selection criteria, 23 articles were included as case-control studies that assessed the relationship between 23 polymorphisms in DNMT1 and cancer risk were included. The rs2228612, rs2228611, rs16999593, and rs10420321 single nucleotide polymorphisms (SNPs) were most frequently investigated. The rs2228612 co-dominant model [P=0.037, OR=0.89, 95% CI (0.80-0.99)] and rs2228611 dominant model [P<0.001, OR=1.32, 95% CI (1.14-0.53)] revealed a substantial association with cancer risk. Subgroup analysis showed an association between the rs2228612 co-dominant [P=0.022, OR=0.58, 95% CI (0.74-0.98)] and recessive [P=0.046, OR=1.15, 95% CI (1.00-1.32)] models with gastrointestinal cancer and the rs2228611 co-dominant model with breast cancer [P=0.024, OR=1.15, 95% CI (1.02-1.29)].

Conclusion: Although the current study found a role for DNMT1 polymorphisms in cancer risk, further high-quality studies are needed to validate these findings.

Objective: This study aimed to develop a hybrid model for variable selection in high-dimensional survival analysis using a support vector regression (SVR), to identify prognostic biomarkers associated with survival in oral cancer (OC) patients through the analysis of gene expression data.

Materials and methods: In this retrospective cohort study, gene expression profiles (54,613 probes) related to 97 patients from the GSE41613 dataset from the GEO repository were used. First of all, martingale residuals were obtained using a Cox regression without covariates, and were used as pseudo-survival outcome. Then, the particle swarm optimization (PSO) and genetic algorithm (GA) were used in combination with SVR for selecting features related to pseudo-survival outcome. Concordance index (C-index), mean absolute error (MAE), mean squared error (MSE) and R-squares, were used to evaluate the performance of the models using selected features. Functional enrichment analysis was performed using DAVID database, and external validation utilized three independent datasets (GSE9844, GSE75538, GSE37991, GSE42743).

Results: The findings indicated that the PSO-based method outperformed the GA-based method, achieving a smaller MAE (0.061) and MSE (0.005), R-square (0.99) and C-index (0.973), selecting 291 probes from 1069 screened. A protein-protein interaction (PPI) network was constructed, including 200 nodes and 120 edges. Eleven key genes with the highest degree, including RBM25, SMC3, PRPF40A, POLE, SRRT, BCLAF1, PDS5B, HNRNPR, JAK1, MED23, and SULT1A1 were identified as significant biomarkers associated with OC survival.

Conclusion: The PSO-based hybrid model effectively improved SVR performance in survival prediction for OC patients and identified key prognostic biomarkers. Despite its promising results and validation on independent datasets, limitations in generalizability and signs of overfitting suggest the model is not yet ready for clinical use. Further studies with larger, diverse datasets are recommended.

Objective: The accumulation of amyloid plaques and disturbance of cholesterol homeostasis are implicated in the pathophysiology of Alzheimer's disease. Apolipoprotein E (ApoE) and cholesterol 24-hydroxylase (CYP46A1) are key proteins involved in the efflux and metabolism of excess cholesterol, and small non-coding RNAs (miRNAs), can help to regulate the expression of the genes encoding these proteins. The aim of the present study was to investigate the alterations in the expression of APOE and CYP46A1 genes, as well as their respective regulatory miRNAs, in astrocytes treated with amyloid beta (Aβ).

Materials and methods: In this experimental study, isolated astrocyte cells were cultured and treated with Aβ for 24 hours. Changes in the expression of APOE and CYP46A1 genes, as well as their regulating miRNAs, were assessed using the realtime polymerase chain reaction (PCR) technique.

Results: The expression of APOE and CYP46A1 genes increased with Aβ treatment. MiR-33a-5p, as the negative regulator of the APOE gene exhibited significant decrease. Additionally, miR-let-7a-5p, as the positive regulator of the APOE gene, showed an increase in the Aβ treated group. Moreover, miR-98-5p, as the negative regulator of the CYP46A1 gene, showed a half-fold decrease. While, miR-27a-3p as the positive regulator of the CYP46A1 gene, increased significantly with Aβ treatment.

Conclusion: Alterations in the expression of APOE and CYP46A1 genes, as well as the expression of miRNAs regulating these genes, in astrocytes treated with Aβ suggests that the cell is attempting to modify the regulatory pathways of cholesterol homeostasis in the brain under pathological conditions, such as Alzheimer's disease.

Objective: Granulosa cell tumor (GCT) is a sex cord-stromal rare malignancy; and is associated with infertility. Extracellular vesicles (EVs) are small secreted vesicles containing proteins, mRNA, and miRNA, therefore modulating signaling pathways potentially in recipient cells and in this way, they can help cancer spread through intercellular communication. In this research, the capability of ovarian cancer-derived EVs for inducing proliferation and metastasis in GCs is investigated.

Materials and methods: In the experimental study, EVs were isolated by ultracentrifugation from A2780 human ovarian cancer cell-conditioned. Mouse GCs were mechanically isolated from 8 female mice. ovarian cancer-derived EVs were then added to the GCs (experimental groups: untreated GCs (control group), GCs treated with concentrations of 10, 25, and 50 μg/ml), and cell viability, migration, apoptosis, and estrogen hormone measurements were assessed by MTT, scratch assay, propidium iodide (PI) staining, annexin-V-FITC/PI and ELISA assay respectively. Gene's expression of Amh, Foxl2, Gdf-9, and Igf-1r were determined using real-time polymerase chain reaction (PCR).

Results: GCs treated with ovarian cancer EVs indicate an increase in cell viability compared to the control cells (P<0.05). Also, ovarian cancer EVs showed an increase in the migration potency. The other results this experimental study showed that ovarian cancer EVs increase in the estrogen secretion level (P<0.001) and reduce the apoptosis of GCs compared to the control group. It is further showed that EVs isolated from ovarian cancer (A2780 cell line) can induce the RNA expression level of several genes in recipient GCs, such as Amh, Foxl2, and Igf-1r (P<0.01, P<0.001), respectively, while the RNA expression level of Gdf-9 gene was decreased in comparison with the control.

Conclusion: To sum up, the research here provides a novel insight into the role of ovarian cancer EVs in proliferation and metastasis in GCs and prevention of infertility caused by granulosa cancer.

Isoflavones are phytoestrogen compounds that can regulate the growth of prostate cancer (PCa) cells. While most studies indicate phytoestrogens mediate their biological effects by activating estrogen receptors (ERs) ERα and ERβ, recent research suggests that they also activate the G-protein coupled ER (GPER). This study aimed to investigate the effects of genistein and daidzein on cell migration and invasion processes in LNCaP and PC3 human PCa cell lines. Furthermore, the role of GPER as a mediator of the effects of these two isoflavones was evaluated using the specific receptor antagonist G15. LNCaP and PC-3 were pre-treated with G15 (1 μM) before phytoestrogens (50 μM) treatments. Following treatment, cell lines were cultured in Matrigel-uncoated Transwell inserts for migration assays and coated inserts for invasion assays. Treatments with genistein and daidzein significantly decrease cell migration and invasion in LNCaP and PC-3. Additionally, the GPER antagonist G15 suppresses the effects of genistein and daidzein, restoring the migratory and invasive capacities in both cell lines. Based on the results, we suggest that GPER regulates cell migration and invasion of LNCaP and PC-3 mediated by genistein and daidzein. Further research is needed to elucidate the signaling pathways involved to evaluate its role and potential as a therapeutic intervention target in PCa.