This is the time to thank all the CytoJournal peer reviewers for their critical participation in the exercise of publishing scientific literature prior to its dissemination. The critical efforts of all the peer reviewers are obvious by the continued high standards of CytoJournal articles. Although according to the “close review” policy of CytoJournal (http://www.cytojournal.com/prp.asp), all CytoJournal reviewers are anonymous, their deserving role must be acknowledged prominently. This editorial collectively acknowledges the efforts of all CytoJournal reviewers [Tables [Tables11–3] for reviewing the manuscripts submitted during 2008 through 2010. Some of these manuscripts were recommended to be published after appropriate responses by the authors to the reviewer criticism.(1–52) Others could not be accepted for publication based on reviewer recommendations. Table 1 CytoJournal reviewers 2008 Table 3 CytoJournal reviewers 2010 As per the peer review policy (http://www.cytojournal.com/prp.asp), depending on various reasons, including potential of conflict of interest, some of the manuscripts are periodically assigned to an “academic editor” who completes the review process and recommends final decision to the executive editor. The academic editors are acknowledged in the article published.(53,54) However, the academic editors of the manuscripts which were not recommended to be published are also included in this list with all the reviewers. On behalf of CytoJournal editorial board (http://www.cytojournal.com/editorialboard.asp) and Cytopathology Foundation Inc. ( http://www.cytopathology-foundation.org), I thank all the reviewers and academic editors for participating in and completing CytoJournal peer review process for multiple CytoJournal manuscripts from 2008 through 2010 [Tables [Tables11–3].[1–52] Please accept this editorial as token of appreciation for the time and efforts to improve the final quality of CytoJournal articles. We request your continued participation in high standards of CytoJournal peer review process in a timely fashion. This is a critical component of any scientific peer reviewed journal such as CytoJournal.[53,54] Table 2 CytoJournal reviewers 2009 Based on the input from you, CytoJournal is organizing an opportunity to claim CME for participating in peer review of CytoJournal manuscripts. Soon CytoJournal peer reviewers completing high-quality review process in a timely fashion could claim AMA Category 1 CME credits after each review, in collaboration with Wayne State University School of Medicine (Southeast Michigan Center for Education), Detroit, MI, USA. Once this feature becomes active, it will be obvious during the acceptance and completion of review process. In addition, you will be notified as soon as the online infrastructure for claiming the CME is complete. For any manuscript submitted to CytoJournal, the reviewers are selected from the core reviewer panel (http://www.cytojournal.com/reviewers.asp). All the experts in cytopathology are invited to join the ever increasing demand of good reviewers by visiting “Reviewer Corner” on CytoJournal home page (www.cytojournal.com) and selecting “Join as CytoJournal Reviewer” at http://www.cytojournal.com/joinus.asp. CytoJournal and Cytopathology Foundation also thank all the authors for contributing to the “open access charter” of CytoJournal http://www.cytojournal.com/downloads/cpl.pdf. By publishing in CytoJournal, you could disseminate your work freely all over the world without losing your hard earned copyright by agreeing to share it in public domain. The benefits include study proven higher access, visibility, and downloads of your work.[55–57] The probability of citation, which depends on the topic and quality of your study-article, is similar to the conventional journals.[57]
Tumor-associated macrophages (TAMs) are important components of the breast cancer (BC) microenvironment that contributes to tumor progression by secreting cytokines. Chemokine ligand 5 (CCL5) expression is upregulated in BC, but the specific mechanism of TAM-derived CCL5 in BC microenvironment remains elucidated. This article aims to explore the role of TAM-derived CCL5 in the progression of BC. M2-type TAMs were induced from Tohoku Hospital Pediatrics-1 (THP-1) monocytes. CCL5 expression in TAM cells was evaluated. The biological effects of recombinant or TAM-derived CCL5 on ZR-75-30 cells were examined using proliferation, migration, invasion assays, and epithelial-mesenchymal transition (EMT) markers. CCL5 was silenced in TAMs and hypoxia-inducible factor-1α (HIF-1α) was overexpressed to explore the downstream signaling pathway. CCL5 expression was significantly increased in TAMs compared with that in BC cells (P < 0.001). Recombinant CCL5 and TAM-conditioned medium promoted ZR-75-30 cell proliferation, migration, and invasion and EMT (P < 0.01), whereas CCL5 knockdown in TAMs markedly reversed these effects (P < 0.001). EMT-related changes, including decreased E-cadherin expression and increased Vimentin, matrix metalloproteinase (MMP)2, and MMP9 expression levels, were reversed by CCL5 silencing (P < 0.01). Inhibition of CCL5 significantly reduced HIF-1α and vascular endothelial growth factor (VEGF) expression levels (P < 0.001), and these reductions were rescued by HIF-1α overexpression (P < 0.01). Functional rescue experiments confirmed that HIF-1α overexpression restored cell proliferation and invasion and EMT suppressed by CCL5 knockdown. Angiogenesis was suppressed by CCL5 silencing and subsequently restored by HIF-1α overexpression (P < 0.001). TAM-derived CCL5 plays an oncogenic role in BC by regulating HIF-1α/VEGF pathway-mediated malignancy and angiogenesis.
Fine needle aspiration cytology (FNAC) is a standard diagnostic tool and has several advantages which are specific to soft tissue tumors (STT). STT arises from non-epithelial and extra-skeletal tissue of the body, excluding the reticuloendothelial system, glia, and supporting tissues of organs. Primary STTs are very rare and encompass a wide range of different tumor types; therefore, the classification and diagnosis remain complex. If the tumor is diagnosed as benign, surgery may not be necessary. However, in the case of malignancy, a cytological diagnosis allows for the initiation of palliative treatment. This study was conducted to evaluate the role of FNAC in the diagnosis of STT by sub classifying them based on their cytology features and finally correlating their FNAC findings with histopathological results with special reference to immunohistochemistry (IHC) in diagnostic challenging cases. It was a hospital-based retrospective study conducted in a tertiary care center, over a period of 2 years, i.e., from July 2016 to May 2018. After obtaining approval from the ethics committee of the institution, the study was carried out. All STT cases subjected to FNAC followed by incisional, excisional, or core needle biopsy or complete resection for histopathological examination were studied. The slides of these patients were retrieved and reviewed to establish a proper diagnosis. IHC was carried out in most cases whenever there was a diagnostic dilemma. In our study, among 50 patients who presented with STT, 40 were benign, and 10 were malignant. The median age of incidence was 41, and male-to-female ratio was 0.85:1. The most common site was the trunk. There were four discordant cases, and the final evaluation of all results showed sensitivity, specificity, positive predictive value of tests, and negative predictive value of tests of FNAC as 100%, 76.92%, 92.5%, and 100%, respectively. In STT, most were benign tumors, with lipomas being the most common among them. While benign and malignant were easy to diagnose, intermediate spindle cell tumors were often missed on cytology. These intermediate tumors represent the diagnostic "grey zone." With increasing experience, STT can be diagnosed more accurately. FNAC is therefore a standard pre-operative diagnostic tool and a reliable alternative to open biopsy for diagnosing STT.
This study aims to explore the cellular composition and transcriptional variability within the tumor microenvironment (TME) of non-small-cell lung cancer (NSCLC) using single-cell RNA sequencing (scRNAseq) and to assess the role of EF-hand domain-containing protein 2 (EFHD2) in tumor progression and its involvement in relevant signaling pathways. We analyzed scRNA-seq datasets to map the cellular and transcriptional landscape of NSCLC tumors. Immunohistochemistry (IHC) was employed to validate the expression of EFHD2 and assess immune cell infiltration in clinical samples. To further investigate the functional effect of EFHD2, we performed Western blot and quantitative real-time polymerase chain reaction analyses as well as cell proliferation, migration, invasion, and apoptosis assays. We also explored the janus kinase (JAK)-signal transducers and activators of transcription (STAT) signaling pathway as a potential underlying mechanism. The scRNA-seq analysis revealed that epithelial cells were the predominant population within the TME, alongside endothelial cells, fibroblasts, macrophages, and a small proportion of stem cells. EFHD2 expression exhibited considerable variability, with higher levels associated with clusters enriched in transcriptionally active and immunomodulatory pathways. The IHC results demonstrated elevated EFHD2 expression and immune cell infiltration in tumor tissues compared with adjacent non-tumor tissues. EFHD2 expression in the NSCLC TME correlates with immune cell infiltration and may play a significant role in tumor progression and immune modulation. The JAK-STAT signaling pathway may be a potential mechanism underlying the effect of EFHD2. This work provides a new avenue for targeted therapy in NSCLC.
Epilepsy is a neurological disease whose onset causes a variety of sequelae, reducing the standard of living. TWIK-related K+ channel 1 (TREK-1) has been linked to epilepsy. C-X-C motif chemokine receptor (CXCR2) is a potential target for the treatment of epilepsy inflammation. This work aims to observe the effect of CXCR2 expression on neuron autophagy and TREK-1 expression in the hippocampus of epileptic mice. An animal model of epilepsy was established, and the CXCR2 gene was silenced. The expression of TREK-1, interleukin (IL)-1b, tumor necrosis factor-a, and IL-6 in the hippocampus of mice was detected by quantitative real-time polymerase chain reaction. Autophagy-related proteins beclin-1 and microtubule-associated protein light chain 3 (LC3) were examined by Western blot. Cell proliferation and activity were analyzed using cell counting kit-8 and 5-Bromodeoxyuridine assays. Compared with that in the normal group, TREK-1 expression decreased and CXCR2 expression increased significantly in the hippocampus of epileptic model mice (P < 0.01). Two autophagy markers, beclin-1 and LC3 II/LC3 I, showed an increased expression in the hippocampal regions of the epileptic model group (P < 0.01). In addition, B-cell lymphoma 2 (Bcl-2)-associated X protein levels increased and Bcl-2 levels decreased in the epileptic mice (P < 0.01). After CXCR2 silencing, the expression of proinflammatory factor in the hippocampus of epileptic mice significantly decreased (P < 0.01). In vitro, cell viability and proliferation increased significantly after silencing CXCR2 (P < 0.05). Meanwhile, the expression levels of TREK-1 and Bcl-2 significantly increased (P < 0.001) and the levels of autophagy markers decreased in vivo and in vitro (P < 0.01). In vivo, CXCR2 expression did not change significantly after silencing TREK-1. After silencing TREK-1 and overexpressing CXCR2, the proliferation ability of HT22 cells decreased significantly (P < 0.001). Epileptic mice's hippocampal neuronal damage can be ameliorated by CXCR2 suppression. One possible explanation is that epileptic mice's hippocampus tissues express more TREK-1, which prevents excessive neuronal autophagy and lowers apoptosis.
CD30 positivity serves as a critical diagnostic hallmark for anaplastic lymphoma kinase positive anaplastic large cell lymphoma (ALK+ ALCL); however, rare cases with partial or complete loss of CD30 expression present diagnostic challenges. We report a unique case with two morphologically and immunophenotypically distinct subpopulations within the same tumor: A typical CD30-positive component and a CD30-negative counterpart. Comparative genetic analysis revealed overlapping but divergent molecular features, including nucleophosmin 1-ALK (NPM1-ALK) fusion, notch receptor 1 mutations, and differential gene amplifications involving MYC proto-oncogene and BCL2-like 1. These findings offer insights into the regulatory mechanisms of CD30 and highlight that loss of CD30 expression alone should not preclude the diagnosis of ALK-positive ALCL, underscoring the importance of integrated histopathological and molecular evaluation in diagnostically challenging cases. Meanwhile, it provides a basis for optimizing the diagnostic process of clinically suspected cases.
Colorectal cancer (CRC) is a malignancy known for its aggressive behavior and notable mortality burden. Indoleamine 2,3-dioxygenase 1 (IDO1), an immune checkpoint molecule, is markedly upregulated in CRC, and it has been implicated in the regulation of key components of the ferroptosis pathway. This study aimed to elucidate the regulatory interaction between IDO1 and ferroptosis and assess the potential therapeutic effect of combined IDO1 inhibition with ferroptosis inducers Erastin and RSL3 in CRC. Three CRC cell lines (CT26, MC38, and HT-29) and one normal colon epithelial cell line (NCM460) were treated with ferroptosis inducers to evaluate the changes in IDO1 expression through reverse transcription quantitative polymerase chain reaction (RT-qPCR) and Western blotting. MC38 cells with stable IDO1 knockdown were generated through lentiviral transduction. The expressions of ferroptosis-related genes following IDO1 silencing were assessed through RT-qPCR and Western blot. The antitumor efficacy of combined IDO1 knockdown and Erastin/RSL3 treatment was evaluated through cell counting kit-8, Transwell migration, and wound-healing assays. Enzyme-linked immunosorbent assay was employed to quantify intracellular levels of glutathione (GSH) and malondialdehyde (MDA). Reactive oxygen species (ROS) levels were measured via flow cytometry, and ferrous ion (Fe2+)and nicotinamide adenine dinucleotide phosphate (NADPH) concentrations were determined through colorimetric analysis. Treatment with Erastin and RSL3 upregulated IDO1 expression in CRC cells. Silencing of IDO1 in MC38 cells resulted in increased expressions of cyclooxygenase-2, acyl-CoA synthetase long chain family member 4, and NADPH oxidase 1, alongside decreased expression of GSH peroxidase 4, solute carrier family 7 member 11, ferritin heavy chain 1, and nuclear factor E2-related factor 2. The combination of IDO1 knockdown with Erastin/RSL3 treatment significantly reduced the proliferation, invasion, and migration of MC38 cells (P < 0.001). This combined treatment also elevated intracellular MDA, ROS, and Fe2+ levels while lowering GSH and NADPH levels. Combination therapy with IDO1 inhibitor and ferroptosis inducer may be an effective way to improve treatment efficacy in CRC.
The objectives of the study are to develop and validate a novel prognostic model for extranodal natural killer/T-cell lymphoma (ENKTCL) by integrating clinical and pathological parameters. We retrospectively analyzed 106 patients with ENKTCL (2008-2020) from the Department of Pathology of Yantai Yuhuangding Hospital and the Affiliated Hospital of Nantong University, constructing the novel international prognostic index (NIPI) model through multivariable Cox regression of immunohistochemistry (IHC) markers (age, MTP53, Ki-67, lactate dehydrogenase, hemoglobin, platelet-tolymphocyte ratio) and quantitative dot blot (QDB) tumor microenvironment features. The model demonstrated significant risk stratification (P < 0.001) with a 3-year area under the curve of 0.72 (IHC) and 0.80 (QDB), outperforming Ann Arbor staging (P > 0.05) and existing international prognostic index (P = 0.00036)/natural killer lymphoma prognostic index (P = 0.00017) models. The QDB-based implementation showed superior prognostic discrimination (P = 0.00014), highlighting its potential for precise individualized therapy. NIPI provides improved risk stratification for ENKTCL, and QDB-based analysis offers enhanced precision in individualized therapy. This model addresses the unmet needs for ENKTCL prognostication and warrants further multicenter validation.
Breast cancer (BC) is one of the most common malignant tumors among women. Gamma-aminobutyric acid (GABA) is abnormally expressed in various cancers, but its effect on BC remains unclear. This study aims to explore the expression changes of glutamic acid decarboxylase 1 (GAD1) in BC and its mechanism of promoting tumor occurrence and development by regulating GABA synthesis. GAD1 expression and GABA levels in BC cells (MDA-MB-231) and normal breast epithelial cells (MCF-10A) were measured. GAD1 overexpression and knockdown cell lines were constructed to evaluate the changes in GABA levels and their effects on cell proliferation and invasion ability. Macrophage M2 polarization and protumor factor levels were analyzed by coculturing tumor cells with macrophages. Cell activity was detected by coculturing with CD8+ T cells, and tumor immune responses were examined by immunofluorescence and cytokine detection. In vitro tube formation was used to simulate angiogenesis, and angiogenesis ability was detected. Western blot was used to analyze the expression of vascular endothelial cell markers, and calcium-tracer dyes were used to detect calcium ion (Ca2+) influx in endothelial cells. GAD1 expression and GABA level in MDA-MB-231 cells were significantly higher than those in normal cells (P <0.01). GAD1 overexpression enhanced cell proliferation, invasion, and migration abilities and simultaneously promoted macrophage polarization to M2, releasing protumor factors interleukin-10 and transforming growth factor-beta and inhibiting the antitumor effect of M1-type macrophages (P <0.05). GAD1 inhibition had the opposite effect (P <0.01). GAD1 overexpression also suppressed the activity of CD8+ T cells; decreased the expression of programmed death-1 in CD8+T cells and the levels of interferon-gamma, perforin, and granzyme B; weakened the immune response; and promoted tumor angiogenesis (P <0.05). The number of in vitro vascular lumen formation increased, the expression of vascular markers rose, and Ca2+ influx was enhanced (P <0.01). Reverse operation demonstrated that GABA promoted the initiation and advancement of BC by regulating the immune microenvironment and angiogenesis (P <0.05). GAD1 is highly expressed in BC. Increasing GABA synthesis promotes tumor cell proliferation and metastasis, regulates the immune microenvironment toward immunosuppression, and enhances tumor angiogenesis. This work reveals the crucial role of the GAD1-GABA pathway in BC and provides a potential therapeutic target for this disease.
The Yokohama system standardizes fine-needle aspiration cytology (FNAC) reporting for breast lesions, while the breast imaging-reporting and data system (BI-RADS) categorizes lesions based on imaging risk assessment. This study aimed to evaluate the diagnostic accuracy of FNAC using the Yokohama System in correlation with BI-RADS classifications. A retrospective analysis was conducted on 188 breast lesion cases that underwent FNAC and were categorized using the BI-RADS system. The breast FNAs were classified according to the Yokohama System and compared with histopathological diagnoses to calculate the risk of malignancy (ROM). The ROM for the BI-RADS categories was also determined. Sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were calculated for both. According to the Yokohama classification, 5.9%, 52.1%, 4.8%, 8.5%, and 28.7% were classified as non-diagnostic, benign, atypical, suspicious, and malignant, respectively. The majority of cases were classified as BIRADS 3 (23.9%) and 5 (23.9%), followed by 4a (22.3%). Higher Yokohama and BI-RADS categories were more prevalent among malignant cases, with 64.9% in Yokohama category 5 and 51.9% in BI-RADS 5. Sensitivity was highest in Yokohama Scenario C (81.8%), while specificity (96.4%) and PPV (92.6%) were highest in Scenario A. When BI-RADS 4b, 4c, 5, and 6 were considered malignant, Scenario C had the highest sensitivity (87.0%) and NPV (90.4%), while Scenario B demonstrated the highest diagnostic accuracy (87.7%). FNAC using the Yokohama System and imaging-based BI-RADS classification both show high diagnostic accuracy for breast lesions. While their individual performance is comparable, further studies are needed to explore whether combining these modalities improves diagnostic outcomes in clinically indeterminate cases.
Cancer stem cells are involved in chemotherapy resistance. Neuroblastoma (NB) is a common solid tumor in children, which is responsible for about 15% of pediatric cancer deaths. Jumonji domain-containing 6 (JMJD6), which is an arginine demethylase and lysine hydroxylase, regulates the progression of various tumors. However, the molecular mechanism underlying JMJD6's activity in NB stem cells remains unclear. Thus, this study aims to explore the role of JMJD6 in NB stem cells. Side population and sphere formation assays were used to investigate the role of JMJD6 in NB stem cell expansion. Immunohistochemistry was performed to determine the expression level of JMJD6 in NB tissues. Luciferase reporter assays were used to discover the effect of JMJD6 on the wingless/integrated (WNT) pathway. JMJD6 was upregulated in NB tissues and cells (P < 0.05). NB patients with high JMJD6 levels had poor outcomes. JMJD6 could serve as an independent prognostic factor for NB. JMJD6 overexpression promoted NB stem cell expansion, whereas JMJD6 knockdown had the opposite effect, as determined by cell and animal models. Mechanism Roche studies showed that JMJD6 promoted NB stem cell expansion by activating the WNT pathway. JMJD6 promotes NB stem cell expansion by activating the WNT pathway, thereby providing a new target for NB therapy and a prognostic factor for patients with NB.
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Breast cancer continues to be a leading and aggressive cancer in women. Despite improvements in early treatment, challenges such as rapid tumor proliferation, metastasis, and drug resistance persist. This research examines how the deubiquitinase ubiquitin-specific protease 5 (USP5) maintains Forkhead box M1 (FOXM1) protein stability and its impact on the advancement of breast cancer. Data from The Cancer Genome Atlas were utilized to investigate the expression patterns and interactions of USP5 and FOXM1 in breast cancer. The breast cancer cell lines were subjected to functional testing, including invasion, migration, and proliferation. The expression and interaction of USP5 and FOXM1 were examined using quantitative real-time polymerase chain reaction, Western blot, and co-immunoprecipitation analyses. Protein stability and ubiquitination assays were performed to evaluate the effect of USP5 on FOXM1 stability. USP5 stabilized the FOXM1 protein by deubiquitination. Overexpression of USP5 increased FOXM1 levels, while USP5 knockdown accelerated FOXM1 degradation. The deubiquitinating enzyme USP5 inhibited the proteasomal degradation of FOXM1, enhancing its stability. Functional assays showed that USP5 overexpression promoted breast cancer cell progression, while USP5 knockdown inhibited these malignant phenotypes. In vivo analysis showed that FOXM1 knockdown reduced tumor volume, and USP5 overexpression with FOXM1 knockdown increased tumor size. The findings suggest that USP5 promotes breast cancer progression by regulating FOXM1 stability. USP5 enhances breast cancer progression by stabilizing FOXM1 through deubiquitination.
This study aimed to analyze the prevalence, genotype distribution, and infection burden of human papillomavirus (HPV) as well as their association with cytological findings, among 7,601 women attending a tertiary referral center in Athens, Greece. This retrospective, single-center cross-sectional study analyzed cervical samples from women attending a specific institution (2019-2024) that underwent polymerase chain reaction-based HPV genotyping, while cytological findings were classified according to the Bethesda system. Statistical analyses assessed the relationships between HPV types, infection burden, cytological outcomes, and age groups. Overall HPV prevalence was 31%, including 18.4% single and 12.6% multiple infections. The most frequent HPV genotypes detected were HPV-42, HPV-51, HPV-16, HPV-53, and HPV-54. Multiple infections were more common in younger women, while single infections predominated with increasing age. Cytological results showed 82.3% normal, 15.9% low-grade squamous intraepithelial lesions (LSILs), 1.3% atypical squamous cells of undetermined significance (ASC-US), and 0.5% high-grade squamous intraepithelial lesions (HSILs). HPV types 16, 18, 31, 33, 42, and 51 were strongly associated with LSIL and/or HSIL. Women with multiple HPV infections had higher odds of ASC-US and LSIL compared to those with single infections; however, the opposite pattern was observed for HSIL. Age was inversely associated with ASC-US and LSIL but not with HSIL. These findings highlight the importance of HPV genotype and infection burden in cervical disease risk in this patient cohort and underscore the need for targeted screening and vaccination strategies tailored to the local epidemiology.
Cardiac arrest followed by resuscitation can induce brain injury, and currently, there are no effective treatments for brain damage after cardiopulmonary resuscitation (CPR), necessitating the exploration of additional therapeutic strategies and prevention approaches. This study aimed to investigate the mechanism of action by which SA4503 activates the Sigma-1 receptor (Sig-1R) to protect against ischemic brain injury in both in vitro and in vivo models. The goal of this study was to provide theoretical support for SA4503 as a potential therapeutic agent and promote clinical intervention research for post-resuscitation brain injury following CPR. This study explored the mechanism underlying the ability of Sig-1R activation to mitigate brain injury following cardiac arrest and resuscitation in rats through both in vivo and in vitro models. The methods used include enzyme-linked immunosorbent assay, magnetic resonance imaging, Western blot analysis, and flow cytometry. The in vivo results demonstrated that the Sig-1R agonist SA4503 significantly attenuated neurological deficits in rats subjected to CPR. In vitro mechanistic investigations revealed that SA4503 potently reversed Sig-1R protein downregulation, reduced apoptosis, ameliorated mitochondrial dysfunction, and reduced endoplasmic reticulum (ER) stress in the oxygen-glucose deprivation/reperfusion (OGD/R) group. This study further confirms that Sig-1R activation confers protective effects against brain injury following cardiac arrest and resuscitation, as well as against OGD/R-induced injury in HT22 cells. The underlying mechanism involves the mitigation of apoptosis driven by mitochondrial dysfunction and ER stress.
Integrin β1 (ITGB1) reportedly participates in osteoblast differentiation, mineralization, and migration. Nevertheless, its role and underlying mechanism in osteoblast differentiation and migration during acute bone loss after fracture are not yet clear. This research was designed to measure the role of ITGB1 in osteoblast differentiation and migration and the underlying mechanism. ITGB1 expression was assessed in MC3T3-E1 cells at different incubation times (0, 4, 7, 14, 21, and 28 days) through quantitative real-time polymerase chain reaction. Alkaline phosphatase (ALP) activity determination, ALP staining, and Alizarin red staining were performed to evaluate the differentiation degree of osteoblasts. Western blot was performed to detect the expression of markers related to osteoblast differentiation. Osteoblast migration ability was determined by wound healing and transwell assays. The molecular mechanism by which ITGB1 modulated the differentiation and migration of osteoblasts was examined by Western blot. ITGB1 expression increased significantly after 14, 21, and 28 days of osteoblast differentiation. ITGB1 increases enhanced osteoblast differentiation and migration; conversely, reduced ITGB1 inhibits osteoblast differentiation and migration. Mechanically, ITGB1 facilitated the activation of extracellular signal-regulated kinase 1/2 (ERK1/2) signaling pathway, and the suppression of the ERK ERK1/2 pathway attenuated the effects of ITGB1 on osteoblast differentiation and migration. ITGB1 plays an important role in osteoblast differentiation and migration by activating the ERK1/2 signaling pathway, which may provide novel insights into bone injury treatment.
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Myocardial infarction (MI) is linked to high mortality, which highlights the need for early diagnosis and intervention to prevent heart failure. N6-methyladenosine (m6A) methylation of ribonucleic acid (RNA) influences endothelial dysfunction and MI. Here, the effects and associated mechanisms of fat mass and obesity-associated gene (FTO) on vascular endothelial cell (EC) function, as well as myocardial damage in mice following MI were investigated. A human umbilical vein EC (HUVEC) model of oxygen-glucose deprivation (OGD) as well as a mouse MI model were used to detect MI-induced endothelial and myocardial damage. EC function was examined using cell-counting-kit-8, wound healing, migration, and tube formation assays. Methylated RNA immunoprecipitation (MeRIP) sequencing and RNA sequencing were utilized to analyze the m6A modifications and RNA expression patterns in control and FTO-overexpressing ECs following OGD. FTO and ATP-binding cassette subfamily F member 1 (ABCF1) levels were evaluated using western blotting. The relationship between FTO and ABCF1 was determined using RNA immunoprecipitation-quantitative polymerase chain reaction (RIPqPCR), MeRIP-qPCR, and RNA stability assays. Echocardiography and Masson's trichrome and hematoxylineosin staining were used to measure myocardial injury. FTO expression was reduced in infarcted myocardial tissue and OGD-induced HUVECs (P < 0.01). Functionally, FTO overexpression improved the impaired function of OGD-treated HUVECs and restored myocardial function in MI mice (P < 0.01). Mechanistically, FTO reduced m6A methylation of ABCF1 mRNA and raised ABCF1 levels in OGD-treated HUVECs (P < 0.01). Taken together, our study revealed that FTO overexpression restored OGD-induced endothelial dysfunction and myocardial pathological injury after MI. FTO increased ABCF1 expression in OGD-treated HUVECs through an m6A-dependent mechanism. These findings offer new insights into mitigating EC dysfunction and myocardial injury after MI.
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