Search Results (170)

Colorectal cancer (CRC) remains a leading cause of cancer-related mortality worldwide, characterized by a multi-step carcinogenesis process involving genetic mutations and epigenetic alterations. Despite advances in screening and therapy, challenges such as treatment resistance, recurrence, and metastasis persist. Emerging evidence highlights the critical role of epigenetic modifications, particularly N7-methylguanosine (m7G), in post-transcriptional regulation. This ubiquitous RNA modification participates extensively in tumor biological behaviors by regulating RNA stability, processing, and translation. Studies have shown that dysregulation of m7G modification is closely associated with adverse clinical outcomes in CRC. This review systematically summarizes the biological functions of m7G modification and its key regulatory proteins (such as METTL1/WDR4, eIF4E, etc.), with a focus on their roles in the pathogenesis, progression, prognosis, and diagnosis of, as well as therapy for, colorectal cancer. m7G modification and related molecules hold potential as novel biomarkers and therapeutic targets, thereby providing new strategies for the precision diagnosis and treatment of colorectal cancer. Full article

Morphine addiction is driven in part by persistent reward-associated memory, yet the molecular mechanisms linking translational control to cellular stress responses remain unclear. In the present study, using a mouse morphine-induced conditioned place preference and N2a cell model, we investigated apoptosis-related alterations in the medial prefrontal cortex and the involvement of eIF4E-dependent translational regulation and BDNF/TrkB signaling. Morphine-induced conditioned place preference was associated with an increase in TUNEL-positive cells in the medial prefrontal cortex, accompanied by upregulation of Bax and downregulation of Bcl-2. In N2a cells, morphine induced apoptosis in a dose-dependent manner. Morphine also increased neuronal eIF4E expression in both mPFC tissue and N2a cells, accompanied by upregulation of BDNF and TrkB. Inhibition of the eIF4E/eIF4G interaction with 4EGI-1 significantly affected morphine-induced CPP formation and altered apoptosis-related markers and BDNF/TrkB expression. Notably, intra-mPFC administration of 4EGI-1 suppressed morphine-induced CPP without affecting anxiety-like behavior, locomotor activity, or general learning and memory performance. These findings suggest that eIF4E-dependent translational regulation is functionally associated with morphine-induced reward memory and apoptosis, potentially in association with changes in BDNF/TrkB molecular expression. This study offers novel insight into the molecular basis of morphine addiction and highlights a potentially targetable translational regulatory pathway for therapeutic intervention. Full article

Objectives: The study aimed to evaluate the effects of full-fat rice bran (FFRB; Tainung No. 81, Taiwan) at various doses on insulin resistance, muscle atrophy, and gut microbiota composition in middle-aged ovariectomized (OVX) mice fed a high-fat diet (HFD), using young sham-operated mice as a life-stage reference group. Methods: Thirty-six female ICR mice were assigned to six groups, including OVX mice fed HFD with or without 5%, 10%, or 20% FFRB. Results: Compared with HFD-fed OVX controls, 20% FFRB reduced body weight gain by 43%, decreased visceral fat mass, and improved insulin resistance (homeostasis model assessment of insulin resistance, HOMA-IR reduced by 65%, P_trend = 0.001). FFRB attenuated the decline in relative grip strength (forelimb, P_trend = 0.013; four-limb, P_trend < 0.001), and upregulated muscle protein synthesis genes, including insulin receptor substrate 1 (IRS-1), mammalian target of rapamycin (mTOR), eukaryotic translation initiation factor 4E binding protein 1 (eIF-4EBP1), while downregulating forkhead box protein O1 (FOXO1), muscle RING-finger protein-1 (MuRF-1), and interleukin (IL)-6. FFRB was also associated with higher fecal acetate levels (P_trend < 0.001), upregulated colonic tight junction genes (occludin and zonula occludens (ZO)-1), and greater relative abundance of g_Muribaculum. Correlation analyses revealed positive associations between short-chain fatty acids (SCFAs) and muscle strength, muscle anabolic markers, genus Lachnospiraceae_UCG_001, and Muribaculum. Conclusions: Dietary inclusion of FFRB was associated with favorable metabolic and muscle-related parameters in HFD-fed middle-aged OVX mice, with potential involvement of gut microbiota and SCFA alterations. Full article

Plants perceive light signals through photoreceptors such as CRY1 to regulate growth and development. It is well-known that Arabidopsis CRY1 is a nucleocytoplasmic protein that mediates light inhibition of hypocotyl elongation in the nucleus, but the mechanisms by which CRY1 regulates root growth and functions in the cytoplasm remain poorly understood. Here, we identified eIF3G1, a subunit of the eukaryotic translation initiation factor 3 (eIF3) complex, as a CRY1-interacting protein associated with light-regulated root development. Under blue light, eif3g1 mutants showed longer primary roots, whereas eIF3G1 overexpression reduced root elongation, accompanied by corresponding changes in root apical meristem size. Differential irradiation experiments indicated that shoot illumination is required for eIF3G1-dependent root phenotypes. Transcriptome analysis revealed changes in translation-related and light-responsive genes in response to eIF3G1 perturbation. Comparison with the cry1 transcriptome revealed overlapping differentially expressed genes, including BIC1 and BIC2, and the bic1 bic2 double mutant showed reduced root elongation. Together, these findings identify eIF3G1 as a CRY1-interacting factor that contributes to the shoot-dependent regulation of root growth under blue light, suggesting that eIF3G1 may be associated with the CRY1-dependent shoot-to-root regulation of root growth. Full article

Although sorafenib (SOR) is effective for advanced hepatocellular carcinoma (HCC), significant metabolic heterogeneity limits its therapeutic effect. In this study, we employed high-resolution matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI) to profile the spatial lipidomic alterations in 3D HepG2 spheroids following SOR treatment. Interestingly, sphingophospholipid and glycerophospholipid metabolism played crucial roles. In an orthotopic HCC mouse model, immunohistochemical and immunofluorescence staining confirmed that SOR induced immunological and inflammatory changes. Moreover, transcriptomic and Q-PCR analyses showed increased expression of Stat1, Zbp1, Parp14, Irf1, and Tifa along with decreased Eif4e2 in the SOR treatment group compared to the tumor control group. Bio-layer interferometry and molecular docking data also indicated that ZBP1 possessed favorable binding affinities with SOR. Overall, our findings demonstrated that SOR dramatically disrupted sphingolipid metabolism in tumor cell spheroids and, in an orthotopic model, activated the NOD-like receptor signaling pathway, accompanied by altered secretion of inflammatory factors and macrophage polarization. These results suggest that SOR exerts dual effects on tumor cell lipid metabolism and the tumor immune microenvironment. These findings provide a conceptual basis for future exploration of lipid-modulating therapeutic strategies in HCC. Full article

Programmed cell death 4 (PDCD4) protein is a tumour suppressor protein that inhibits mRNA translation by inhibiting RNA helicase, eukaryotic initiation factor 4A (eIF4A). We have previously reported that PDCD4 interacts with the internal ribosome entry site (IRES) element of B-cell lymphoma extra-large (Bcl-xL) mRNA and inhibits its IRES-mediated translation initiation. S6 kinase (S6K)-mediated phosphorylation of PDCD4 activates its degradation and derepresses IRES-mediated translation initiation of Bcl-xL mRNA. eIF3F (one of the subunits of eIF3 complex) was reported to recruit S6K to phosphorylate eIF3G. Therefore, we investigated the possibility of co-regulation of PDCD4 and eIF3F by S6K and the regulation of IRES-mediated translation initiation by PDCD4–eIF3F. Here, we demonstrated that PDCD4 interacts with several subunits of eIF3. Specifically, eIF3F directly interacts with PDCD4 in an RNA-independent manner. Depletion of PDCD4 in glioblastoma (GBM) cells resulted in decreased levels of certain eIF3 subunits, including eIF3F. Additionally, depletion of eIF3F from GBM cells decreased the levels of PDCD4 protein. We also showed that PDCD4 and eIF3F directly interact with Bcl-xL RNA independently of each other. By performing IRES reporter, polysome profiling assays and EMSA we have demonstrated that eIF3F regulates IRES-mediated translation of Bcl-xL mRNA, likely via its interaction with PDCD4. Full article

Background: Translational regulation constitutes a critical layer of gene expression control in plants, yet the contribution of endogenous 5′ untranslated regions (5′ UTRs) to translational efficiency remains incompletely defined. While viral and synthetic leader sequences have been widely used to enhance protein production, comparatively few native plant 5′ UTRs have been systematically characterised. The objective of this study was to identify and functionally evaluate endogenous plant 5′ UTR elements that promote translation through post-transcriptional mechanisms. Methods: A 79-nucleotide fragment (CYSTM α) derived from the 5′ UTR of Arabidopsis thaliana CYSTM1 (AT1G05340) was cloned upstream of reporter genes and assessed using dual-luciferase assays in transient expression systems (Nicotiana benthamiana and A. thaliana) and in stable transgenic Arabidopsis lines. Translational activity was further evaluated in monocot wheat germ extract and in Escherichia coli. Transcript abundance was quantified by qRT-PCR. Publicly available ribosome profiling and m⁶A datasets were analysed to assess translational efficiency and RNA modification status. Results: In N. benthamiana and A. thaliana, CYSTM α increases reporter protein production 3–7 fold relative to the control and 30–130% above the benchmark Tobacco Mosaic Virus (TMV) Ω leader, without altering mRNA abundance. The CYSTM α sequence also enhances luciferase translation in monocot wheat germ extract and elevates translation 5-fold in E. coli. CYSTM α contains three motifs that may promote translation, namely three CAA repeats that are associated with translation initiation, an AMAYAA motif that is associated with eIF3 binding, and two N6-adenosine DRACH sites that are associated with cap-independent translation. Additionally, ribosome profiling revealed high translational efficiency (TE = 3.25) of native CYSTM1. Conclusions: CYSTM α represents a compact endogenous 5′ UTR element that enhances translation across multiple experimental systems. These findings expand the repertoire of plant-derived translational enhancers and provide insight into sequence features associated with efficient mRNA translation in plants. Full article

Cigarette smoke (CS) is the primary risk factor for the development of chronic obstructive pulmonary disease (COPD). Investigating the impact of CS on human airway epithelium is important for understanding COPD development and combating its effects. While some studies show that long exposure to CS activates inflammasome formation in airway epithelium, leading to cytokines’ maturation and release, its acute effect on inflammation regulation requires further elucidation. Due to the importance of acute cellular responses in modulating cell survival and controlling inflammatory outcomes, we examined the effect of acute cigarette smoke extract exposure on human bronchial epithelial cells. Due to the high reactive oxygen species content in CS, we hypothesize that acute CS exposure activates the integrated stress response (ISR) pathway leading to stress granules (SG) formation to facilitate oxidative stress resolution and promote cell survival. Immunostaining, fluorescence confocal imaging, quantitative analyses, and immunoblotting were performed to test our hypothesis. We report here that acute exposure to CS extract triggers canonical SG formation by activating the ISR pathway via the PERK/eIF2α arm in a reactive oxygen species-dependent manner. SG formation is abolished upon inhibiting PERK or eIF2α function, or by scavenging oxidants prior to smoke exposure. Characterizing SG formation in terms of measuring SG size and abundance and the sequestration of the SG marker G3BP1 reveals that SG formation is maximal at 15% CS extract exposure for 2 h and undergoes gradual disassembly at longer exposure times. This is closely dependent on cytoplasmic p-eIF2α levels. These results demonstrate that acute exposure to CS activates the protective ISR pathway to potentially reduce the detrimental effects of CS and promote stress resolution and cell survival. Full article

Euonymus bungeanus is a highly valued ornamental tree/shrub species widely utilized in landscaping and afforestation in Northeast Asia, yet molecular studies on this species remain limited due to the lack of validated reference genes for reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR). In this study, 16 candidate reference genes were selected based on classical plant reference genes and our previous transcriptome data. Their expression stability was comprehensively evaluated using 64 samples collected from diverse tissues and plants subjected to various abiotic stress/hormone treatments across multiple time points. Across all samples analyzed, PBG1 (20S proteasome beta subunit G1) exhibited the highest overall expression stability, followed by VAPD (vacuolar ATP synthase subunit D) and EIF4A (eukaryotic translation initiation factor 4A). For tissue-specific analysis, TSR2 (pre-rRNA-processing protein), VAPD, and PBG1 demonstrated the greatest stability. Under specific stress conditions, PBG1 and EIF4A were identified as the most stable genes under low- and high-temperature conditions. PP2A (protein phosphatase 2A) and TUB6 (beta-6 tubulin) were optimal for drought stress, while TSR2, SRP (nuclear speckle splicing regulatory-like protein), and PBG1 exhibited superior stability under salt stress. These findings establish a validated panel of reference genes enabling accurate and reliable gene expression normalization in E. bungeanus, thereby facilitating future functional genomics studies in this economically and ecologically important species. Full article

Background/Objectives: Saikosaponin D (SSD) is a bioactive compound from traditional Chinese herbs with known anti-tumor activities, including apoptosis induction, autophagy modulation, and inhibition of cell migration and invasion. However, the mechanisms underlying its effects on human endometrial cancer Ishikawa cells remain elusive. This study aimed to investigate the anti-tumor effects of SSD on EC Ishikawa cells and elucidate the molecular pathways involved, focusing on DNA damage, cell cycle regulation, autophagy, endoplasmic reticulum (ER) stress, and AMPK signaling. Methods: We performed in vitro experiments using Ishikawa cells and in vivo studies using a female BALB/c nude mouse xenograft model. DNA damage was assessed via comet assay, intracellular Ca²⁺ concentration via Fluo-3 AM staining, autophagy via transmission electron microscopy, and apoptosis via flow cytometry. Autophagy was inhibited using 3-methyladenine, and ER stress was modulated with the PERK inhibitor GSK2656157. Protein expression levels of related genes were analyzed by western blotting. No preregistration number or CONSORT details applied, as this was a pre-clinical study. Results: SSD treatment was associated with DNA damage and G2/M phase cell cycle arrest in Ishikawa cells both in vitro and in vivo. SSD was associated with an increased LC3II/LC3I ratio and activation of the AMPK pathway. It was also associated with ER stress, as evidenced by downregulation of PERK, mTOR, and eIF2α, and upregulation of p-eIF2α. Furthermore, SSD was associated with modulation of the AMPK signaling pathway to inhibit cell migration and invasion. Conclusions: SSD exerts anti-tumor effects on human EC Ishikawa cells in vitro and in vivo through mechanisms involving DNA damage, G2/M arrest, autophagy, ER stress, and AMPK-mediated inhibition of migration and invasion. These findings suggest that SSD may represent a potential therapeutic agent for EC. Full article

Foot-and-mouth disease virus (FMDV) is a highly contagious picornavirus that affects cloven-hoofed animals and carries significant economic implications for the global livestock industry. FMDV features two Leader (L) protein isoforms, Lab and Lb, differing at their amino termini by 28 amino acids (La region). Currently, the activity of La protein sequences has not been investigated. To address this issue, the comparison study of biological and functional roles of Lab and Lb was performed as the La region alone did not independently perform protein function. We found that Lab and Lb significantly regulated FMDV replication and pathogenicity, and their coexistence afforded optimal FMDV properties. Subsequently, we observed that both L isoforms cleaved eukaryotic translation initiation factor 4G (eIF4G) I, suppressed type I and type III interferon (IFN) expression, and exhibited marked cytotoxicity, indicating that they were all key components in FMDV’s antagonism of host antiviral defenses. Finally, the subcellular distribution of Lab and Lb was detected. Despite dual localization in cytoplasmic and nuclear compartments, both isoforms displayed different spatial distribution patterns, and Lb induced more pronounced morphological changes to host cells than Lab. Furthermore, bioinformatics predicted that the La region might contain a non-classical secretory signal peptide, potentially facilitating Lab distribution to the cell membrane or extracellular space. Collectively, the primary encoding role of La region was to control the intracellular distribution of L protein, as opposed to regulating its functional activity. This study may help to deepen our understanding of why FMDV encoded two isoforms of L protein. Full article

Addressing principal challenges in common bean (Phaseolus vulgaris L.) breeding requires a holistic approach. A combined strategy was implemented to assess seven genotypes (landraces and commercial varieties) for yield potential, stability and resistance to bean common mosaic virus (BCMV) under Mediterranean low-input conditions. Pure-line selection and prognostic breeding together with SSR and CAPS-SCAR marker-assisted selection (MAS) formed the core methodology. Significant variation was detected across 24 morpho-agronomic descriptors, while SSR revealed 48.57% polymorphic loci and private alleles in specific landraces. High genetic coefficients of variation and high heritability were recorded for yield-related traits. Phenotypical evaluation showed diverse responses to BCMV, with mild symptoms predominating (52.14%). Entries G1 (45%) and G5 (35%) exhibited the highest frequency of the symptomless resistant phenotype. Molecular screening at I and bc-3/eIF4E loci confirmed G5’s robust dominant I gene profile, while G1 included individuals carrying both the dominant I gene and recessive bc-3, offering a valuable source for pyramiding resistance. Additionally, G1 (LI = 2.35; 100%) performed strongly in productivity, whereas G2 (SI = 3.1; 100%) and G7 (SI = 2.8; 89.7%) exhibited exceptional stability. Overall, the mixed-model approach highlighted the complementary characteristics of the tested genotypes and identified G1, G2, G5 and G7 as promising candidates for future breeding programs targeting high yield, low-input adaptability and resistance to BCMV. Full article

Background: Oral Submucous Fibrosis (OSMF) is a significant global oral health problem, particularly prevalent in India, with a high risk of progression to Oral Squamous Cell Carcinoma (OSCC). This study investigates the molecular mechanisms involved in the transformation of OSMF to OSCC using transcriptomic profiling. Methods: High-throughput RNA sequencing was performed on fresh de novo OSCC samples (n = 8) and OSMF derived OSCC using Illumina-compatible NEXTflex Rapid Directional RNA Sequencing. Normalization and differential gene expression analysis were conducted, and genes exhibiting an absolute log2 fold change of ≥2 with a co-variate-adjusted p-value ≤ 0.05 were identified as significant. Results: Upregulated genes were associated with cytokine and immune responses (ABRA, TTTY14, EIF1AY), cellular proliferation and apoptosis (LINC00314, RPS4Y1, SERPINA5, TRIM63, FABP7), and energy metabolism, indicating metabolic adaptations during malignant progression. Pathway analysis showed increased expression of TNNT1, TNNI1, MYL4, and ACTN3, implicating muscle development and embryonic pathways in OSMF transformation. Conversely, genes related to epithelial differentiation and keratinization (FLG, FLG2, HRNR, TCHH, KRT73), immune regulation and tumor suppression (HLA-G, UNC5D), and metabolic signaling were downregulated, reflecting loss of tissue integrity and immune control. Conclusions: OSMF-derived OSCC exhibits a distinct transcriptomic landscape compared with de novo OSCC, characterized by altered epithelial differentiation, immune modulation, and activation of developmental pathways. The observed gene dysregulation findings establish that OSCC developing in the background of OSMF is molecularly distinct from de novo OSCC, underscoring the biological impact of the pre-existing fibrotic milieu on tumor transcriptional architecture. Full article

Influenza A virus (IAV) remains a major global threat, highlighting the need for host-targeted antiviral strategies. While some probiotics offer prophylactic protection, their therapeutic potential post-infection is poorly understood. Here, we investigated human-derived Lactobacillus brevis ZG2488 for its antiviral potential against IAV. Strikingly, a more pronounced reduction in viral titer was observed when live bacteria were administered therapeutically post-infection, compared to preventive pretreatment. Transcriptomic analysis suggested that the therapeutic effect of viable bacteria was associated with a modulated host response, including the downregulation of specific host factors implicated in viral replication (e.g., KPNA2, NUP98, EIF2S1) and a delayed interferon-beta (IFNB1) induction. In contrast, preventive effects appeared to be mediated by heat-stable components. These findings highlight a viability-dependent mode of action for L. brevis ZG2488 and contribute to the growing evidence that certain probiotics may exert antiviral effects through targeted host modulation rather than solely through broad immune activation. Full article