Enhanced Cell Proliferation Research Articles

Epigenetically associated IGF2BP3 upregulation promotes cell proliferation by regulating E2F1 expression in hepatocellular carcinoma

RNA-binding proteins (RBPs) are a class of proteins that primarily function by interacting with different types of RNAs and play a critical role in regulating the transcription and translation of cancer-related genes. However, their role in the progression of hepatocellular carcinoma (HCC) remains unclear. In this study, we analyzed RNA sequencing data and the corresponding clinical information of patients with HCC to screen for prognostic RBPs. Insulin-like growth factor 2 mRNA-binding protein 3 (IGF2BP3) was identified as an independent prognostic factor for liver cancer. It is upregulated in HCC and is associated with a poor prognosis. Elevated IGF2BP3 expression was validated via immunohistochemical analysis using a tissue microarray of patients with HCC. IGF2BP3 knockdown inhibited the proliferation of Hep3B and HepG2 cells, whereas IGF2BP3 overexpression promoted the expansion of HuH-7 and MHCC97H cells. Mechanistically, IGF2BP3 modulates cell proliferation by regulating E2F1 expression. DNA hypomethylation of the IGF2BP3 gene may increase the expression of IGF2BP3, thereby enhancing cell proliferation in HCC. Therefore, IGF2BP3 may act as a novel prognostic biomarker and a potential therapeutic target for HCC.

Neuromedin S regulates goat ovarian granulosa cell proliferation and steroidogenesis via endoplasmic reticulum Ca2+-YAP1-ATF4-c-Jun pathway.

Neuromedin S (NMS) plays key roles in reproductive regulation, while its function and mechanism in follicular development remain unclear. The current study aims to investigate the specific role and mechanisms of NMS and its receptors in regulating the proliferation and steroidogenesis of ovarian granulosa cells (GCs). Phenotypically, a certain concentration of NMS addition promoted the proliferation and estrogen production of goat GCs, accompanied by an increase in the G1/S cell population and upregulation of the expression levels of cyclin D1, cyclin dependent kinase 6, steroidogenic acute regulatory protein, cytochrome P450, family 11, subfamily A, polypeptide 1, 3beta-hydroxysteroid dehydrogenase, and cytochrome P450, family 11, subfamily A, polypeptide 1, while the effects of NMS treatment were effectively hindered by knockdown of neuromedin U receptor type 2 (NMUR2). Mechanistically, activation of NMUR2 with NMS maintained endoplasmic reticulum (ER) calcium (Ca2+) homeostasis by triggering the PLCG1-IP3R pathway, which helped preserve ER morphology, sustained an appropriate level of endoplasmic reticulum unfoldedprotein response (UPRer), and suppressed the nuclear translocation of activating transcription factor 4. Moreover, NMS maintained intracellular Ca2+ homeostasis to activate the calmodulin 1-large tumor suppressor kinase 1pathway, ultimately orchestrating the regulation of goat GC proliferation and estrogen production through the Yes1 associated transcriptional regulator-ATF4-c-Jun pathway. Crucially, the effects of NMS were mitigated by concurrent knockdown of the NMUR2 gene. Collectively, these data suggest that activation of NMUR2 by NMS enhances cell proliferation and estrogen production in goat GCs through modulating the ER and intracellular Ca2+ homeostasis, leading to activation of the YAP1-ATF4-c-Jun pathway. These findings offer valuable insights into the regulatory mechanisms involved in follicular growth and development, providing a novel perspective for future research.

CALU promotes lung adenocarcinoma progression by enhancing cell proliferation, migration and invasion

BackgroundLung cancer is the second most common cancer with the highest mortality in the world. Calumenin as a molecular chaperone that not only binds various proteins within the endoplasmic reticulum but also plays crucial roles in diverse processes associated with tumor development. However, the regulatory mechanism of calumenin in lung adenocarcinoma remains elusive. Here, we studied the impact of calumenin on lung adenocarcinoma and explored possible mechanisms.Methods5-ethynyl-2’-deoxyuridine assay, colony formation, transwell and wound healing assays were performed to explore the effects of calumenin on the proliferation and migration of lung adenocarcinoma cells. To gain insights into the underlying mechanisms through which calumenin knockdown inhibits the migration and proliferation of lung adenocarcinoma, we performed Gene Ontology, Kyoto Encyclopedia of Genes and Genomes, Gene Set Enrichment Analysis and Ingenuity Pathway Analysis based on transcriptomics by comparing calumenin knockdown with normal A549 cells.ResultsThe mRNA and protein levels of calumenin in lung adenocarcinoma are highly expressed and they are related to an unfavorable prognosis in this disease. Calumenin enhances the proliferation and migration of A549 and H1299 cells. Gene Set Enrichment Analysis revealed that knockdown of calumenin in A549 cells significantly inhibited MYC and V-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog signaling pathways while activating interferon signals, inflammatory signals, and p53 pathways. Ingenuity pathway analysis provided additional insights, indicating that the interferon and inflammatory pathways were prominently activated upon calumenin knockdown in A549 cells.ConclusionsThe anti-cancer mechanism of calumenin knockdown might be related to the inhibition of MYC and KRAS signals but the activation of interferon signals, inflammatory signals and p53 pathways.

Cancer-associated snaR-A noncoding RNA interacts with core splicing machinery and disrupts processing of mRNA subpopulations.

RNA polymerase III (Pol III) activity in cancer is linked to the production of small noncoding (nc)RNAs that are otherwise silent in most tissues. snaR-A (small NF90-associated RNA isoform A) - a hominid-specific ncRNA shown to enhance cell proliferation, migration, and invasion - is a cancer-emergent Pol III product that remains largely uncharacterized despite promoting growth phenotypes. Here, we applied a combination of genomic and biochemical approaches to study the biogenesis and subsequent protein interactions of snaR-A and to better understand its role as a putative driver of cancer progression. By profiling the chromatin landscapes across a multitude of primary tumor types, we show that predicted snaR-A upregulation is broadly linked with unfavorable outcomes among cancer patients. At the molecular level, we unexpectedly discover widespread interactions between snaR-A and mRNA splicing factors, including SF3B2 - a core component of the U2 small nuclear ribonucleoprotein (snRNP). We find that SF3B2 levels are sensitive to high snaR-A abundance and that depletion of snaR-A alone is sufficient to decrease intron retention levels across subpopulations of mRNA enriched for U2 snRNP occupancy. snaR-A sensitive genes are characterized by high GC content, close spatial proximity to nuclear bodies concentrated in pre-mRNA splicing factors, and functional enrichment for proteins involved in deacetylation and autophagy. We highlight examples of splicing misregulation and increased protein levels following snaR-A depletion for a wide-ranging set of factors, suggesting snaR-A-driven splicing defects may have far-reaching effects that re-shape the cellular proteome. These findings clarify the molecular activities and consequences of snaR-A in cancer, and altogether establish a novel mechanism through which Pol III overactivity may promote tumorigenesis.

Suppressing SENP1 inhibits esophageal squamous carcinoma cell growth via SIRT6 SUMOylation.

Esophageal squamous cell carcinoma (ESCC) is a prevalent tumor in the gastrointestinal tract, but our understanding of the molecular mechanisms underlying ESCC remains incomplete. Existing studies indicate that SUMO specific peptidase 1 (SENP1) plays a crucial role in the development and progression of various malignant tumors through diverse molecular mechanisms. However, the functional mechanism and clinical implications of SENP1 in the progression of ESCC remain unclear. Bulk RNA-Sequencing (RNA-seq) was used to compare potential genes in the esophageal tissues of mice with ESCC to the control group. The up-regulated SENP1 was selected. The protein level of SENP1 in ESCC patient samples was analyzed by immunohistochemistry and western blot. The potential prognostic value of SENP1 on overall survival of ESCC patients was examined using tissue microarray analysis and the Kaplan-Meier method. The biological function was confirmed through in vitro and in vivo knockdown approaches of SENP1. The role of SENP1 in cell cycle progression and apoptosis of ESCC cells was analyzed by flow cytometry and western blot. The downstream signaling pathways regulated by SENP1 were investigated via using RNA-Seq. SENP1-associated proteins were identified through immunoprecipitation. Overexpression of Sirtuin 6 (SIRT6) wildtype and mutant was performed to investigate the regulatory role of SENP1 in ESCC progression in vitro. Our study discovered that SENP1 was upregulated in ESCC tissues and served as a novel prognostic factor. Moreover, SENP1 enhanced cell proliferation and migration of ESCC cell lines in vitro, as well as promoted tumor growth in vivo. Thymidine kinase 1 (TK1), Geminin (GMNN), cyclin dependent kinase 1(CDK1), and cyclin A2 (CCNA2) were identified as downstream genes of SENP1. Mechanistically, SENP1 deSUMOylated SIRT6 and subsequently inhibited SIRT6-mediated histone 3 lysine 56 (H3K56) deacetylation on those downstream genes. SIRT6 SUMOylation mutant (4KR) rescued the growth inhibition upon SENP1 depletion. SENP1 promotes the malignant progression of ESCC by inhibiting the deacetylase activity of SIRT6 pathway through deSUMOylation. Our findings suggest that SENP1 may serve as a valuable biomarker for prognosis and a target for therapeutic intervention in ESCC.

Physical and biological study of Yttrium oxide/titanium oxide/carbon nanotube/polystyrene films for wound healing purpose

Different contributions of dopants based on Yttrium oxide (Y2O3)/titanium(IV) oxide (TiO2)/carbon nanotube (CNT)/have been incorporated into the polymeric matrix of Polystyrene (PS). These films were fabricated in films due to their feasibility and low cost. The contact angle revealed that the dopants supported the hydrophilicity of the films that Y2O3/TiO2/CNT/PS represents the second highest wettability with (56.15 ± 6.35°) films, while Y2O3/PS film displays rough surface with the highest wettability with (32.5 ± 0.9°). we examined the impact of Y2O3/TiO2/CNT/PS composite on the viability of a human lung cell-line to better understand its biological behavior. Specifically, we measured cell viability as a means to assess the effect of the composite. The results revealed that at a concentration of 5000 μg/ml, the cell viability was measured to be 31.8 %. However, at a lower concentration of 2.4 μg/ml, the cell viability increased significantly to 117.9 %. These findings highlight a concentration-dependent relationship between the composite and the viability of the human lung cell-line. The observed cell viability exceeding 100 % indicates that the Y2O3/TiO2/CNT/PS composite scaffold had a positive effect on cell proliferation and generation. This suggests that the modified PS scaffold is conducive to cellular growth and can be considered safe for human use. The results imply that the composite scaffold promotes a favorable cellular environment, potentially supporting tissue regeneration and indicating its potential suitability for biomedical applications. The surface morphology shows that the topography of the films was changed clearly with the dopants of Yttrium oxide, titanium(IV) oxide and CNT components. The TiO2 granules implanted in the PS composition cause an increase in pore size to an average of 2.2 μm. Pore diameters ranging from 100 to 500 μm may enhance cell proliferation, while lower sizes lead to more stable implantable discs.

GINS2 promotes the progression of human HNSCC by altering RRM2 expression.

GINS2 exerts a carcinogenic effect in multiple human malignancies, while it is still unclear that the potential roles and underlying mechanisms of GINS2 in HNSCC. TCGA database was used to screen out genes with significant differences in expression in HNSCC. Immunohistochemistry and qRT-PCR were used to measure the expression of GINS2 in HNSCC tissues and cells. GINS2 was detected by qRT-PCR or western blot after knockdown or overexpression. Celigo cell counting, MTT, colony formation, and flow cytometric assay were used to check the ability of proliferation and apoptosis. Bioinformatics and microarray were used to screen out the downstream genes of GINS2. GINS2 in HNSCC tissues and cells was up-regulated, which was correlated with poor prognosis. GINS2 gene expression was successfully inhibited and overexpressed in HNSCC cells. Knockdown of GINS2 could inhibit proliferation and increase apoptosis of cells. Meanwhile, overexpression of GINS2 could enhance cell proliferation and colony formation. Knockdown of RRM2 may inhibit HNSCC cell proliferation, while overexpression of RRM2 rescued the effect of reducing GINS2 expression. Our study reported the role of GINS2 in HNSCC for the first time. The results demonstrated that in HNSCC cells, GINS2 promoted proliferation and inhibited apoptosis via altering RRM2 expression. Therefore, GINS2 might play a carcinogen in HNSCC, and become a specific promising therapeutic target.

Membrane Palmitoylated Protein 2 Serves as a Potential Biomarker for Colon Adenocarcinoma through Promoting CGAS-STING Pathway

This research investigated the role of Membrane Palmitoylated Protein 2 (MPP2) in colon adenocarcinoma (COAD), the third most prevalent form of colorectal cancer. Utilizing TCGA, GEPIA, and HPA datasets alongside Western blotting, we examined MPP2 expression in normal versus cancerous tissue. Kaplan-Meier plots from GEPIA indicated lower MPP2 levels in COAD patients correlating with worse overall survival (P < 0.05). Moreover, MPP2 was identified as a distinct prognostic indicator for COAD and significantly associated with clinicopathological features (P < 0.05), assessed via chi-square tests and Cox regression models. We further explored MPP2’s link to COAD immune cells using Cibersort, revealing its correlation with the infiltration of 8 immune cells (P < 0.05). In vivo and in vitro experiments demonstrated that elevated MPP2 levels might enhance cell proliferation and migration while suppressing apoptosis. Applying Gene Set Enrichment Analysis (GSEA), we predicted MPP2’s involvement in tumor progression via the DNA sensing pathway, a finding supported by cellular studies. In summary, COAD patients exhibit decreased MPP2 expression, which inversely correlates with prognosis. MPP2 appears to influence immunosuppression and tumor progression through various mechanisms, suggesting its potential as an independent prognostic marker in COAD. Further investigation into MPP2’s implications is warranted.

The role of light emitting diode in wound healing: A systematic review of experimental studies.

Wounds represent a growing global issue demanding increased attention. To expedite wound healing, technologies are under development, and light emitting diode (LED) devices of varying wavelengths are being explored for their stimulating influence on the healing process. This article presents a systematic literature review aiming to compile, organize, and analyze the impacts of LED devices on wound healing.This review is registered on the PROSPERO platform [CRD42023403870]. Two blinded authors conducted searches in the Pubmed, Web of Science, Scopus, Embase, and ScienceDirect databases. In vitro and in vivo experimental studies assessing LED utilization in the wound healing process were included.The search yielded 1010 studies, of which 27 were included in the review. It was identified that LED stimulates different healing pathways, promoting enhanced cell proliferation and migration, angiogenesis stimulation, increased collagen deposition, and modulation of the inflammatory response.Thus, it can be concluded that the LED stimulates cellular and molecular processes contingent on the utilized parameters. The effects depend on the standards used. Cell migration and proliferation were better influenced by green and red LED. The extracellular matrix components and angiogenesis were regulated by all wavelengths and the modulation of inflammation was mediated by green, red, and infrared LEDs.

Long non-coding RNA MIMT1 promotes retinoblastoma proliferation via sponging miR-153-5p to upregulate FGF2

With the rapid development of biotechnology, long non-coding RNAs (lncRNAs) have shown promising potential for cancer treatment and may become novel therapeutic targets. This study aimed to explore the roles of lncRNAs in retinoblastoma (RB). It involves analysing differentially expressed lncRNAs in RB and normal tissues from the GSE111168 and GSE125903 datasets, further validating them in RB cells. Our findings determined that lncRNA MIMT1 was upregulated in RB cell lines and tissues. In WERI-Rb1 and Y79 cells, silencing MIMT1 significantly inhibited cell proliferation, whereas MIMT1 overexpression enhanced cell proliferation. Furthermore, in vivo xenograft experiments demonstrated that MIMT1 knockdown suppressed tumour volume and weight. Subsequent mechanistic investigations showed that MIMT1 upregulates fibroblast expression of FGF2 by binding to miR-153-5p, ultimately promoting RB cell proliferation. This suggest that MIMT1 functions as an oncogene in RB and potentially serves as a molecular marker for diagnostic and prognostic assessments. Thus, the MIMT1/miR-153-5p/FGF2 pathway is a promising therapeutic target for RB treatment.

Additive manufacturing of polylactic acid scaffolds dip-coated with polycaprolactone for bone tissue engineering

Bone tissue engineering aims to create scaffolds that support bone regeneration, addressing the needs of approximately 1.71 billion people with bone structure problems worldwide. This study explores the fabrication and characterization of 3D-printed polylactic acid (PLA) scaffolds dip-coated with polycaprolactone (PCL), producing complex geometries with interconnected pores, specifically RoundBar Sphere and RoundBar Cube. The scaffolds were then coated with PCL solutions of 0.5 % and 1 % concentrations, applying up to three layers. Surface topology analysis indicated that PCL coating slightly reduced pore size (1150 µm to 900) while improving coverage and integrity. After coating, Fourier Transform Infrared Spectroscopy (FTIR) confirmed the presence of PCL on scaffold surfaces (characteristic bands at 1726, 1175 and 728 cm−1), whose a better coverage was obtained with more layers and higher concentrations of PCL. Coated scaffolds showed not significant change in compressive strengths (2–12 MPa), remaining suitable for trabecular bone applications. Hemolysis assays of the 3D scaffolds promoted non-hemolytic properties (0 % hemolysis), ensuring their blood compatibility. Metabolic activity (>70 %) and live/dead cell assays in human dermal fibroblasts (HDF) exhibited biocompatibilities for all samples, with coated scaffolds promoting enhanced cell proliferation compared to uncoated ones. Additionally, osteoblast metabolic activity (>90 %) and osteoblasts scratch assay demonstrated coated scaffolds promoted an area reduction of 1.36 and 1.53-fold higher than the control group and uncoated scaffold, respectively. In short, these coated scaffolds are promising candidates for bone tissue engineering and bone repair applications.

Enhanced chondrogenic potential in GelMA-based 3D cartilage model via Wnt3a surface immobilization

Cartilage tissue engineering aims to develop functional substitutes for treating cartilage defects and osteoarthritis. Traditional two-dimensional (2D) cell culture systems lack the complexity of native cartilage, leading to the development of 3D regenerative cartilage models. In this study, we developed a 3D model using Gelatin Methacryloyl (GelMA)-based hydrogels seeded with Y201 cells, a bone marrow mesenchymal stem cell line. The model investigated chondrogenic differentiation potential in response to Wnt3a stimulation within the GelMA scaffold and validated using known chondrogenic agonists. Y201 cells demonstrated suitability for the model, with increased proteoglycan content and upregulated chondrogenic marker expression under chondrogenic conditions. Wnt3a enhanced cell proliferation, indicating activation of the Wnt/β-catenin pathway, which plays a role in cartilage development. GelMA hydrogels provided an optimal scaffold, supporting cell viability and proliferation. The 3D model exhibited consistent responses to chondrogenic agonists, with TGF-β3 enhancing cartilage-specific extracellular matrix (ECM) production and chondrogenic differentiation. The combination of Wnt3a and TGF-β3 showed synergistic effects, promoting chondrogenic differentiation and ECM production. This study presents a 3D regenerative cartilage model with potential for investigating cartilage biology, disease mechanisms, and drug screening. The model provides insights into complex cartilage regeneration mechanisms and offers a platform for developing therapeutic approaches for cartilage repair and osteoarthritis treatment.

Involvement of multiple epigenetic mechanisms by altered DNA methylation from the early stage of renal carcinogenesis before proliferative lesion formation upon repeated administration of ochratoxin A

Ochratoxin A (OTA) is a rat renal carcinogen that induces karyomegaly and micronuclei in proximal tubular epithelial cells (PTECs). We previously performed comprehensive gene profiling of alterations in promoter-region methylation and gene expression in PTECs of rats treated with OTA for 13 weeks. The OTA-specific gene profile was obtained by excluding genes showing expression changes similar to those upon treatment with 3-chloro-1,2-propanediol, a renal carcinogen not inducing karyomegaly. In this study, we validated the candidate genes using methylated DNA enrichment PCR and real-time RT-PCR, and identified Gen1, Anxa3, Cdkn1a, and Osm as genes showing OTA-specific epigenetic changes. These genes and related molecules were subjected to gene expression and immunohistochemical analyses in the PTECs of rats treated with OTA, other renal carcinogens, or non-carcinogenic renal toxicants for 4 or 13 weeks. Cdkn1a upregulation and increase of p21WAF1/CIP1+ karyomegalic PTECs were observed with OTA, matching the findings associated with micronucleus-inducing carcinogens. This suggested that the increase of p21WAF1/CIP1+ karyomegalic PTECs is linked to micronucleus formation, which in turn accelerates chromosomal instability. The upregulation of Cdkn1a-related genes with OTA suggests the acquisition of a senescence-associated secretory phenotype, which promotes the establishment of a carcinogenic environment. Meanwhile, OTA specifically caused a decrease of GEN1+ PTECs reflecting Gen1 downregulation and an increase of ANXA3+ PTECs reflecting Anxa3 upregulation, as well as Osm upregulation. OTA may efficiently disrupt pathways for repairing the DNA double-strand breaks that it itself causes, via Gen1 downregulation, and enhance cell proliferation through the upregulation of Anxa3 and Osm. This may exacerbate the chromosomal instability from the early stage of OTA-induced renal carcinogenesis before proliferative lesions form. OTA may cause renal carcinogenesis involving multiple epigenetic mechanisms.

CeO2-loaded PVA/GelMA core-shell nanofiber membrane to promote wound healing

Chronic wounds are a significant health issue due to various pathogenic abnormalities that hinder the healing process. Nanofiber membranes, with their favorable conductivity for cell growth, proliferation, and adhesion, offer immense potential for effective chronic wound healing. In this study, we utilized the coaxial electrospinning method to synthesize core-shell structured nanofiber membranes for chronic wound healing. To obtain superior nanofiber membranes, we modified the shell part of the electrospun nanofiber by combining gelatin methacrylate (GelMA) with cerium oxide (CeO2). The results show that these nanofibers exhibit promising wound healing properties, including ideal hydrophilicity, swelling ratio, porosity, mechanical properties, antibacterial activity, and fibroblast cell viability. With the addition of 3 % w/v CeO2 nanoparticles (PGCe-3 CL sample), the nanofiber membrane had a 53.7° water contact angle, 384.47 % swelling ratio, 83.91 % porosity, and 8.49 MPa tensile strength, which are optimal for the wound healing process. The PGCe-3 CL sample also demonstrated increased antibacterial activity, reducing bacterial colonies by up to 88.46 % for S. aureus and 93.14 % for E. coli compared to PG-CL and PGCe-1.5 CL. Furthermore, the cell viability test showed that the nanofiber membranes are non-toxic to human dermal fibroblast cells and significantly enhance cell proliferation compared to other samples. Accordingly, the synthesized nanofiber membranes loaded with nano CeO2 in this study show promising performance as suitable dressings for chronic wounds. Moreover, the findings of this study provide valuable insights for developing more effective and innovative materials for chronic wound healing.

Prospects and limitations of cumate-inducible lentivirus as a tool for investigating VEGF-A-mediated pathology in diabetic retinopathy

Diabetic retinopathy (DR) is a multifactorial disease displaying vascular-associated pathologies, including vascular leakage and neovascularization, ultimately leading to visual impairment. However, animal models accurately reflecting these pathologies are lacking. Vascular endothelial growth factor A (VEGF-A) is an important factor in the development of micro- and macro-vascular pathology in DR. In this study, we evaluated the feasibility of using a cumate-inducible lentivirus (LV) mediated expression of vegf-a to understand DR pathology in vitro and in vivo. Retinal pigment epithelial cells (ARPE-19) were transduced with cumate-inducible LV expressing vegf-a, with subsequent analysis of vegf-a expression and its impact on cell proliferation, viability, motility, and permeability. Cumate tolerability in adult Wistar rat eyes was assessed as an initial step towards a potential DR animal model development, by administering cumate via intravitreal injections (IVT) and evaluating consequent effects by spectral domain optical coherence tomography (SD-OCT), flash electroretinography (fERG), ophthalmic examination (OE), and immunohistochemistry. Transduction of ARPE-19 cells with cumate-inducible LV resulted in ~ 2.5-fold increase in vegf-a mRNA and ~ threefold increase in VEGF-A protein secretion. Transduced cells displayed enhanced cell proliferation, viability, permeability, and migration in tube-like structures. However, IVT cumate injections led to apparent retinal toxicity, manifesting as retinal layer abnormalities, haemorrhage, vitreous opacities, and significant reductions in a- and b-wave amplitudes, along with increased microglial activation and reactive gliosis. In summary, while cumate-inducible LV-mediated vegf-a expression is valuable for in vitro mechanistic studies in cellular drug discovery, its use is not a feasible approach to model DR in in vivo studies due to cumate-induced retinal toxicity.

Wilms' Tumor 1-Associating Protein Promotes Nonsmall-Cell Lung Cancer Through the Expression of Carcinoembryonic Antigen-Related Cell Adhesion Molecule 5.

This study aimed to analyze the functional roles and molecular mechanism of Wilms' tumor 1-associating protein (WTAP) in the tumorigenesis of nonsmall-cell lung cancer (NSCLC). Retrospective analysis was used. Tumor tissues and surrounding nontumor tissues of 150 patients with NSCLS who were surgically resected in the Fourth Hospital of Hebei Medical University from January 2016 to January 2018 were selected. The expression of WTAP in NSCLC tissues was detected by immunohistochemistry. Clinicopathologic parameters were then subjected to univariate and multivariate Cox regression analysis in purpose of uncovering the independent risk factors for overall survival time. MTS (3-[4,5-dimethylthiazol-zyl]-5-[3-carboxymethoxyphenyl]-2-[4-sulfophenyl]-2H-tetrazoliuzolium, inner salt) assay, colony formation assay, and transwell assays were performed to estimate cell proliferation, migration, and invasion. Meanwhile, the relationship between WTAP and the cell migration and invasion marker-related proteins were evaluated by Western blot analysis and RT-qPCR. WTAP expression was knocked-down in cell lines by shRNA, and RNA-Seq was performed to investigate the pathways regulated by WTAP. In NSCLC patients, WTAP was highly expressed in tumor tissues and the higher expression was significantly associated with poor overall survival (OS) ( P <0.01). Compared with the control group in vitro, the overexpression of WTAP could significantly promote cell proliferation, migration, and invasion ( P <0.01), while knock-down WTAP significantly reduces the above effects ( P <0.01). In a mouse orthotopic implantation model, higher WTAP abundance could significantly promote tumor enlargement compared with the control group ( P <0.01). Compared with the control group, the knock-down of WTAP significantly inhibit the expression of carcinoembryonic antigen-related cell adhesion molecule 5 (CEACAM5) in cell lines ( P <0.01). Besides, in NSCLC, knocked-down CEACAM5 significantly reduced the impact of WTAP on cell proliferation, migration, and invasion compared with the control group ( P <0.05). This study suggests that high expression of WTAP was associated with poor clinical outcomes. CEACAM5 may play a synergistic role with WTAP to jointly promote NSCLC progression by enhancing cell proliferation, invasion, and migration.

DIPG-30. ELUCIDATING DOWNSTREAM MEDIATORS OF ONCOGENESIS INFOXR2-EXPRESSING DIFFUSE MIDLINE GLIOMAS

Abstract BACKGROUND Diffuse midline gliomas (DMGs) are universally fatal pediatric tumors with a median survival of less than one year. Forkhead Box R2 (FOXR2) is a forkhead family TF that we have found to be aberrantly expressed in DMGs. FOXR2 expression is sufficient to enhance glioma formation, and we have shown that regulation of FOXR2 occurs through a previously unrecognized epigenetic mechanism. FOXR2 is necessary for proliferation in FOXR2-expressing DMGs. It is also a potent oncogene, sufficient to enhance cell proliferation and in vivo tumor formation. However, the downstream mediators of FOXR2-mediated gliomagenesis are poorly understood. METHODS We applied an integrative approach using proteomics, transcriptomics, in vitro cancer models, and in vivo in utero electroporation mouse models to systematically evaluate how FOXR2 mediates gliomagenesis. RESULTS We have found that FOXR2 is aberrantly activated in DMGs. FOXR2 enhances MYC protein stability, even in the presence of cycloheximide. However, FOXR2 also exerts its oncogenic effects through MYC independent functions. FOXR2 is highly enriched at E26-transformation (ETS) motifs and specifically activates ETS transcriptional gene sets. To identify FOXR2 protein-protein interactions, we performed immunoprecipitation and mass spectrometry to identify FOXR2 protein interactors. Moreover, we have performed both proteomic and phospho-proteomic analyses of FOXR2-expressing human neural stem cells. These proteomic studies have allowed us to identify proteins, phospho-sites, and signaling pathways that are highly enriched in FOXR2-expressing cells. These pathways could be potential therapeutic targets in FOXR2-expressing DMGs. CONCLUSIONS Taken together, this study elucidates how FOXR2 protein interactors mediate oncogenesis in FOXR2-expressing diffuse midline gliomas.

X-Paste improves wound healing in diabetes via NF-E2-related factor/HO-1 signaling pathway

BACKGROUND Diabetic foot ulcers (DFU), as severe complications of diabetes mellitus (DM), significantly compromise patient health and carry risks of amputation and mortality. AIM To offer new insights into the occurrence and development of DFU, focusing on the therapeutic mechanisms of X-Paste (XP) of wound healing in diabetic mice. METHODS Employing traditional Chinese medicine ointment preparation methods, XP combines various medicinal ingredients. High-performance liquid chromatography (HPLC) identified XP’s main components. Using streptozotocin (STZ)-induced diabetic, we aimed to investigate whether XP participated in the process of diabetic wound healing. RNA-sequencing analyzed gene expression differences between XP-treated and control groups. Molecular docking clarified XP’s treatment mechanisms for diabetic wound healing. Human umbilical vein endothelial cells (HUVECs) were used to investigate the effects of Andrographolide (Andro) on cell viability, reactive oxygen species generation, apoptosis, proliferation, and metastasis in vitro following exposure to high glucose (HG), while NF-E2-related factor-2 (Nrf2 ) knockdown elucidated Andro’s molecular mechanisms. RESULTS XP notably enhanced wound healing in mice, expediting the healing process. RNA-sequencing revealed Nrf2 upregulation in DM tissues following XP treatment. HPLC identified 21 primary XP components, with Andro exhibiting strong Nrf2 binding. Andro mitigated HG-induced HUVECs proliferation, metastasis, angiogenic injury, and inflammation inhibition. Andro alleviates HG-induced HUVECs damage through Nrf2 /HO-1 pathway activation, with Nrf2 knockdown reducing Andro’s proliferative and endothelial protective effects. CONCLUSION XP significantly promotes wound healing in STZ-induced diabetic models. As XP’s key component, Andro activates the Nrf2 /HO-1 signaling pathway, enhancing cell proliferation, tubule formation, and inflammation reduction.

The Promising Potential of Cold Atmospheric Plasma Therapies

The outstanding properties and chemistry of cold atmospheric plasma (CAP) are not sufficiently understood due to their relatively complex systems and transient properties. In this paper, we tried to present a detailed review of the applications of CAP in modern medicine, highlighting the biochemistry of this phenomenon. Due to its unique characteristics, CAP has emerged as a promising tool in various medical applications. CAP, as a partially—or fully ionized—gas-retaining state of quasi-neutrality, contains many particles, such as electrons, charged atoms, and molecules displaying collective behaviour caused by Coulomb interactions. CAP can be generated at atmospheric pressure, making it suitable for medical settings. Cold plasma’s anti-microbial properties create an alternative method to antibiotics when treating infections. It also enhances cell proliferation, migration, and differentiation, leading to accelerated tissue regeneration. CAP can also be a powerful tool in anti-tumour therapies, stem cell proliferation, dental applications, and disease treatment, e.g., neurology. It is our belief that this article contributes to the deeper understanding of cold plasma therapy and its potential in medicine. The objective of this study is to demonstrate the potential of this relatively novel approach as a promising treatment modality. By covering a range of various biomedical fields, we hope to provide a comprehensive overview of CAP applications for multiple medical conditions. In order to gain further insight into the subject, we attempted to gather crucial research and evidence from various studies, hopefully creating a compelling argument in favour of CAP therapy. Our aim is to highlight the innovative aspects of CAP therapy where traditional methods may have limitations. Through this article, we intend to provide a convenient reference source for readers engaged in the examination of CAP’s potential in medicine.

Dual-responsive metal polyphenol network nanosheets for diabetic wound healing

Diabetic wound healing poses a significant challenge in the biomedical domain. Inadequate angiogenesis and fibroblast dysfunction are two primary characteristics of diabetic microenvironment. Fibroblast dysfunction is characterized by excessive accumulation of reactive oxygen species (ROS) and fragmented mitochondria. Herein, we developed a microenvironment responsive zinc–gallic acid nanosheets (ZnGA NSs) for diabetic wound healing, which slowly release Zn2+ and gallic acid (GA) in response to low pH and excessive ROS. The released Zn2+ enhanced cell proliferation, migration, and endothelial tube formation. And the released GA activated antioxidant enzyme system via the KEAP1/NRF2 pathway and promoted mitochondrial fusion by upregulating MFN1/2 and OPA1, which ultimately reprograms dysfunctional fibroblast. The application of ZnGA NSs effectively facilitated wound healing in a rat diabetic model with full-thickness cutaneous defects. More importantly, skin wounds treated with ZnGA NSs exhibited enhanced angiogenesis, alleviated inflammation, and improved tissue regeneration. These findings highlight the potential of ZnGA NSs as a promising therapeutic modality for diabetic wound treatment.