Dysregulation of microRNA networks is a hallmark of gastric cancer pathogenesis, but the mechanisms driving early-stage disease remain poorly understood. This study utilized integrative bioinformatics analysis of the Gene Expression Omnibus dataset GSE158315 to identify tumor-suppressive microRNAs in early gastric cancer. We identified hsa-miR-519d-3p as a core downregulated microRNA in early-stage tissues. Functional assays in NUGC-3 and MKN-45 cell lines demonstrated that miR-519d-3p overexpression significantly suppressed cell migration and invasion, whereas its inhibition enhanced these malignant phenotypes. Dual-luciferase reporter assays confirmed that miR-519d-3p directly targets the 3' untranslated region of BECN1 (Beclin-1). Silencing Beclin-1 via siRNA mimicked the effects of miR-519d-3p overexpression, while rescue experiments showed that Beclin-1 knockdown reversed the pro-migratory and pro-invasive effects triggered by miR-519d-3p inhibition. Furthermore, monitoring of autophagic flux using mRFP-GFP-LC3 tandem reporters revealed that miR-519d-3p inhibition enhances autophagy in a Beclin-1-dependent manner. Clinical data analysis from The Cancer Genome Atlas further supported the upregulation of Beclin-1 in gastric cancer and its correlation with aggressive clinicopathological features. In conclusion, our findings establish the miR-519d-3p/Beclin-1 axis as a critical regulator of motility and autophagy in gastric cancer, representing a potential therapeutic target for early intervention.
The genetic regulatory mechanisms underlying key economic traits in Tan sheep remain unclear. To systematically analyze gene expression patterns across multiple tissues in Tan sheep, this study selected 20 male Tan sheep aged 6 months from the same rearing background. Tissue samples were collected from the heart, liver, spleen, lung, kidney, longissimus muscle, perirenal fat, and tail fat, and conducted transcriptomic sequencing to construct a multi-tissue transcriptomic atlas of Tan sheep and analyze gene expression patterns across multiple tissues. Multi-tissue expression profiling analysis indicates that tissues with similar physiological functions exhibit a high degree of clustering in their gene expression patterns. The study identified a total of 8,257 low-variation genes, 1,473 medium-variation genes, and 2,983 high-variation genes. Among these, low-variation genes exhibit highly conserved functions and are primarily enriched in critical biological processes such as RNA transport, protein processing, and the mTOR signaling pathway. Additionally, 1,790 tissue-specific genes were identified, with their enriched pathways closely associated with tissue function. WGCNA analysis identified 16 gene modules highly associated with specific tissues, revealing the core regulatory gene networks in each tissue. RT-qPCR was used to validate SLC22A8, LOC101120535, AICDA, and TAT as tissue-specific genes in the heart, kidney, lung, and muscle, respectively. In summary, the multi-tissue transcriptomic atlas of Tan sheep constructed in this study systematically analyzed the expression characteristics of tissue-specific genes, providing important data support for a deeper understanding of the genetic regulatory networks underlying the formation of economic traits in Tan sheep.
INTRODUCTION:Hepatic ischemia-reperfusion injury (HIRI) is a major clinical challenge with limited therapeutic options. Metformin (Met) has demonstrated protective effects against ischemia-reperfusion (IR) injury in other organs by modulating endoplasmic reticulum (ER) stress, but its role and mechanism in HIRI remain unclear. METHODS:A mouse HIRI model was established following Met pre-treatment, with six mice included in each group. HIRI was assessed by measuring serum injury markers, examining histopathological changes, and evaluating apoptosis. ER stress markers were analyzed by western blot. Molecular docking predicted interactions between Met and ER stress proteins. AML12 hepatocytes under hypoxia/reoxygenation (H/R) were treated with Met, VER-155008 (VER), and/or CDN1163, followed by analysis of key cell parameters: viability, lactate dehydrogenase (LDH) release, apoptosis, and calcium flux. In vitro experiments were performed as three independent biological experiments. RESULTS:Met pre-treatment significantly attenuated IR-induced hepatic injury, evidenced by reduced serum injury markers, histopathological changes, and hepatocyte apoptosis (p < 0.05). Met also suppressed the HIRI-induced upregulation of ER stress proteins, including GRP78, p-PERK, p-IRE1α, and CHOP (p < 0.05). Molecular docking predicted a potential interaction of Met with GRP78. In H/R-induced AML12 hepatocytes, Met, VER, and CDN1163 similarly improved viability and reduced LDH release and apoptosis (p < 0.05). The protective effect of Met was not observed in the presence of VER, while it showed synergistic efficacy with CDN1163. Notably, Met did not markedly reduce H/R-induced cytosolic Ca2+ accumulation but effectively inhibited H/R-induced ER stress activation. CONCLUSIONS:Met protects against HIRI by suppressing ER stress, and this effect is associated with GRP78 functional regulation without obvious correction of cytosolic Ca2+ accumulation. These findings suggest that GRP78-related ER stress regulation may represent a potential mechanism underlying the hepatoprotective effect of Met.
Estrogen receptor alpha (ERα) is central to breast-cancer initiation, progression, and endocrine resistance. This narrative review uses a transparent, non-systematic search of peer-reviewed English-language literature published from 2017 through 2025 to integrate ERα structure and isoforms, genomic and non-genomic signaling, post-translational regulation, tumor-microenvironment interactions, heterogeneity, and therapeutic development. The synthesis distinguishes mechanistic, preclinical, and clinical evidence and highlights areas of uncertainty. ESR1 ligand-binding-domain mutations are rare in untreated primary disease but are enriched after endocrine selection in metastatic disease; emerging oral SERDs, ER-directed PROTACs, and biomarker-matched pathway combinations show heterogeneous results across overall and molecularly defined populations. ER-low disease remains a clinically important category requiring confirmation of pathology, integration of tumor biology and disease setting, and individualized use of endocrine and chemotherapy-based strategies. Overall, therapeutic progress increasingly depends on matching ERα-directed interventions to dynamic biomarkers, resistance mechanisms, treatment history, and tolerability rather than assuming uniform benefit across ER-positive disease.
BACKGROUND:Multidrug-resistant (MDR) ESKAPE pathogens, including Enterococcus faecium, S. aureus, Klebsiella pneumoniae, Acinetobacter baumannii, P. aeruginosa, and Enterobacter spp., pose a critical global health threat. OBJECTIVE:This narrative review evaluates microRNAs (miRNAs) as novel antibacterial agents against MDR ESKAPE, focusing on their mechanisms of action, preclinical efficacy, delivery innovations, and translational barriers. METHODS:PubMed and Google Scholar were searched using terms related to miRNA biology, antibacterial activity, ESKAPE pathogens, and delivery platforms RESULTS: miRNAs exert antibacterial effects through three mechanisms: innate and adaptive immune modulation, regulation of host antibacterial pathways (including antimicrobial peptide production), and direct cross-kingdom bacterial mRNA silencing with biofilm disruption. Preclinical evidence highlights key candidates: let-7b-5p achieved a 90% reduction in P. aeruginosa biofilm and restored aztreonam sensitivity, and miR-101-3p, delivered via DNA tetrahedron nanostructures, suppressed polymicrobial biofilms in cystic fibrosis (CF) models. The Rocket-miR platform identified miRNA candidates across all ESKAPE organisms, including miR-877-5p and miR-3127-5p, which target carbapenem-resistant K. pneumoniae and vancomycin-resistant E. faecium. The core translational challenges include miRNA instability, limited cellular uptake, off-target effects, and undefined regulatory pathways. Nanocarrier-based delivery, exosomal platforms, and machine learning-assisted target prediction offer promising solutions to these challenges. CONCLUSION:Current preclinical evidence suggests that miRNAs hold promise as early-stage candidate antibacterial agents through immunomodulation, host pathway regulation, and direct bacterial gene silencing with biofilm disruption. However, no miRNA-based antibacterial therapy has entered clinical evaluation, and substantial translational barriers, including delivery challenges, off-target effects, and undefined regulatory pathways, must be addressed before clinical application can be considered.
The abundance of a long non-coding RNA (lncRNA) is set by both its synthesis and its decay, yet for nuclear lncRNAs the contribution of decay - and whether decay rate is a determinant of function - has been little explored. I addressed this for NEAT1, the architectural scaffold of paraspeckles. By genome-wide half-life measurement (5'-bromouridine immunoprecipitation chase, BRIC), NEAT1 is a representative short-lived lncRNA at the median of the lncRNA half-life distribution (∼3.3 h), unlike the stable MALAT1. Using inhibitor-free decay measurement and bidirectional perturbation, I show that the core nuclear exosome (EXOSC5, EXOSC2), but not the 5'→3' exonuclease XRN2, the exosome-associated 3'→5' exonuclease EXOSC10, or the deadenylase PARN, sets the NEAT1 decay rate: exosome depletion lengthened the NEAT1 half-life (7.8 → 15.1 h) and raised its abundance; reciprocally, EXOSC5 over-expression shortened the NEAT1 half-life and EXOSC2 over-expression reduced its abundance. The resulting changes in NEAT1 abundance scaled the number and size of paraspeckles without increasing paraspeckle-protein mRNA levels, and changed the expression of candidate PSF/NONO target genes: across three orthogonal NEAT1 manipulations, increased NEAT1 abundance was associated with repression of RAP1A and TCF3 and activation of the NONO-regulated gene GJA1, without changing those mRNAs' own stabilities (under EXOSC5 depletion). These results, which complement the subsequently established exosome-adaptor mechanism of NEAT1 turnover by adding a direct decay-rate measurement and a functional read-out, identify RNA decay rate as a control point - a throttle - on lncRNA function, supported through bidirectional perturbation for NEAT1 in HeLa cells.
Salt stress severely limits maize growth and productivity, yet the molecular mechanisms controlling transcription factor stability during salt stress remain poorly understood. Here, we identified ZmMYC7 as a negative regulator of maize salt tolerance and uncovered its post-translational regulation by ZmMAPK3. ZmMYC7 was rapidly induced by salt stress and localized to the nucleus. CRISPR/Cas9-mediated knockout of ZmMYC7 significantly enhanced salt tolerance, accompanied by reduced membrane damage, lower Na+ accumulation, higher K+ retention, and improved Na+/K+ homeostasis under saline conditions. Mechanistically, yeast two-hybrid, bimolecular fluorescence complementation, pull-down, and co-immunoprecipitation assays demonstrated that ZmMAPK3 physically interacts with ZmMYC7 both in vitro and in vivo. In vitro kinase assays further showed that ZmMAPK3 directly phosphorylates ZmMYC7 at Thr367, while phos-tag analysis revealed that salt stress markedly enhances ZmMYC7 phosphorylation in planta. Cell-free degradation assays demonstrated that phosphorylation at Thr367 accelerates the 26S proteasome-dependent degradation of ZmMYC7, thereby reducing its protein abundance. Together, our findings establish a novel ZmMAPK3-ZmMYC7 regulatory module in which salt-induced phosphorylation promotes proteasomal turnover of ZmMYC7, relieving its negative effect on salt tolerance.
Phosphoribosyl pyrophosphate synthetase (PRPS) produces phosphoribosyl pyrophosphate, which is a key precursor for nucleotide biosynthesis. Mutations in the human PRPS1 gene cause Charcot-Marie-Tooth disease and other neuropathies. However, how PRPS1 dysfunction impairs neuronal and synaptic function remains unclear. In this study, we used Drosophila melanogaster to determine the effects of reduced Prps expression on neurons in vivo. Neuronal knockdown of Prps caused severe locomotor impairment in third instar larvae and in adults, which indicated a critical requirement for Prps in motor output. At the neuromuscular junction, Prps depletion led to a reduced number of synaptic boutons accompanied by enlargement of boutons, disrupted presynaptic active zone organization indicated by decreased Bruchpilot puncta, and loss of Futsch-positive microtubule loops. These findings suggested destabilization of cytoskeletal architecture. These phenotypes were consistently observed using independent RNAi lines. Importantly, neuronal expression of Drosophila Prps or human PRPS1 significantly rescued locomotor activity, synaptic growth, active zone organization, and microtubule structure, which are caused by the expression of the Prps short hairpin RNA. The rescue by Drosophila Prps and human PRPS1 highlights evolutionary conservation of PRPS1 function and suggests Drosophila as a model for analyzing PRPS1-associated neuropathies.
Lung cancer is one of the most prevalent and lethal malignancies worldwide, ranking among the leading causes of cancer-related morbidity and mortality. Uncontrolled cellular proliferation constitutes its central pathological hallmark, underscoring the critical need to identify novel, druggable therapeutic targets. Recent studies have demonstrated that chloride intracellular channel 3 (CLIC3), a multifunctional protein exhibiting both ion channel activity and oxidoreductase function, may play pivotal roles in tumorigenesis. However, its functional significance in lung cancer remains poorly understood. Here we found that CLIC3 was significantly upregulated at both mRNA and protein levels in lung cancer tissues, and its high expression was closely correlated with poor patient prognosis. To elucidate the biological role of CLIC3 in lung cancer pathogenesis, we performed genetic knockdown of CLIC3 in multiple lung cancer cell lines. Silencing CLIC3 markedly impaired tumor cell proliferation, migration, and invasion in vitro. Mechanistically, CLIC3 depletion attenuated chloride efflux and induced substantial accumulation of intracellular reactive oxygen species (ROS). Transcriptomic profiling further indicated that knockdown of CLIC3 may repress lung cancer cell proliferation, at least in part, by inhibiting the C4-dicarboxylate transport pathway. Consistent with these in vitro findings, in vivo xenograft models confirmed that CLIC3 ablation significantly inhibits lung tumor growth. In summary, our study demonstrated that CLIC3 as a critical regulator of lung cancer progression, acting through coordinated modulation of intracellular chloride dynamics and ROS balance. These findings position CLIC3 as a promising prognostic biomarker and a compelling novel therapeutic target for lung cancer.
This study aimed to clarify the functional regulatory links among RPA3, GPR87 and p53 in the tumorigenesis and progression of non-small cell lung cancer (NSCLC). Bioinformatic analysis and Western blot assays were performed to evaluate the expression profiles of RPA3 and GPR87 in NSCLC. CCK-8, EdU, wound healing, Transwell and flow cytometry assays were used to explore the effects of RPA3 knockdown on NSCLC proliferation, migration, invasion, cell cycle and apoptosis. Both pharmacological inhibition and siRNA-mediated knockdown of p53, together with GPR87 overexpression, were applied for in vitro and in vivo functional rescue verification. The results showed that RPA3 and GPR87 were highly expressed in NSCLC tissues and cells. RPA3 silencing inhibited cell proliferation, induced apoptosis and caused S-phase arrest in A549 and H23 cells. Notably, both GPR87 overexpression and genetic/chemical inhibition of p53 effectively rescued the tumor-suppressive phenotypes induced by RPA3 depletion. Cellular and xenograft experiments consistently confirmed that RPA3 modulates NSCLC malignant behaviors through functional phenotypic linkage of GPR87 and p53. In summary, RPA3 exerts oncogenic effects by regulating GPR87 and p53-dependent cellular functions, thereby promoting NSCLC cell proliferation, suppressing apoptosis and facilitating tumor growth.
Anthocyanins are crucial secondary metabolites in apple, which determine the commercial value of the fruit and confer essential protection against environmental stresses. Their biosynthesis is governed by a complex network of structural and regulatory genes, modulated by developmental cues, environmental factors, and phytohormone signaling. Emerging evidence highlights gibberellins (GAs) as pivotal regulators of anthocyanin accumulation in apple. Here, we review the current understanding of the molecular mechanisms underlying gibberellin (GA)-mediated anthocyanin biosynthesis. We specifically highlight the role of DELLA proteins in orchestrating this GA-mediated regulatory network and examine how post‑translational modifications of DELLA proteins, together with their interactions with other signaling components, influence anthocyanin accumulation. Furthermore, we discuss the intricate crosstalk between GA and other hormonal pathways, particularly jasmonic acid and Strigolactone, revealing how these interactions fine‑tune anthocyanin production. We also explore how GA signaling integrates environmental cues, such as light and temperature, to coordinate anthocyanin levels in apple. Finally, we identify key knowledge gaps in the GA-anthocyanin regulatory network and propose promising directions for future research.
The WRKY transcription factors play an important role in regulating plant response to drought stress. The results show that SlWRKY30 acts as a negative regulator of drought tolerance, and that overexpressing SlWRKY30 (SlWRKY30OE) promotes vegetative growth. Moreover, the expression of SlWRKY30 was induced by drought stress, and SlWRKY30OE plants exhibited more severe wilting than WT under drought conditions. SlWRKY30 overexpression led to high relative electrolyte leakage, MDA content, and ROS levels in tomatoes under drought stress. In SlWRKY30OE plants, the activities of SOD, POD, CAT and APX were lower than those of WT plants; further, the expression levels of SlFe-SOD, SlCAT1 and SlcAPX were also lower than in WT plants under drought stress. Further analysis of RNA sequencing indicated that the transcription level of the encoding photosystem I and II genes was almost down-regulated, while the degree of SlWRKY30OE plants downregulation was stronger than in WT plants. Consistently, the Pn and Fv/Fm were lower in SlWRKY30OE plants during drought stress. These results indicate that SlWRKY30 reduces drought resistance in tomato by weakening antioxidant capacity, promoting ROS accumulation and membrane damage, and impairing photosynthetic performance under drought stress.
We report a unique clinical case of a male child presenting with a blended phenotype characterized by intellectual disability, seizures, periventricular nodular heterotopia (PVNH), congenital central hypothyroidism, and multiorgan malformations. Exome sequencing (ES) identified three rare variants: a de novo frameshift variant in SETD5 (g.3-9441593 ;NM_001080517.3:c.812dup; p.(Leu271PhefsTer42)), a de novo missense variant in the WW domain of NEDD4L (NM_015277.6:c.1525C > T; p.(Arg529Cys)), and a maternally inherited nonsense variant in TBL1X (X-9711651-C-T) (NM_005647.4:c.1480C > T; p.(Arg494Ter)). While the likely pathogenic SETD5 and TBL1X variants are consistent with the neurodevelopmental delay and central hypothyroidism respectively, the NEDD4L variant of uncertain significance (VUS) represents a plausible candidate for the PVNH. To our knowledge, this is the first report of such a unique overlap, highlighting the importance of considering multilocus genomic variation in heterogeneous neurodevelopmental presentations.
