Long non-coding RNAs (lncRNAs) are key regulators in hepatocellular carcinoma (HCC) progression, yet the function of lncRNA GAS5 in HCC remains controversial. GAS5 expression was analyzed across multiple public datasets. Overexpression experiments were performed in HCC cells to evaluate GAS5’s biological role. Bioinformatic prediction, luciferase reporter experiments, and rescue assays were used to elucidate the molecular mechanism. Xenograft mice were established to confirm GAS5’s in vivo effects. GAS5 levels were markedly reduced in HCC tissues. Excessive GAS5 suppressed cell viability and colony formation, induced G0/G1 arrest, promoted apoptosis, and inhibited migration and invasion. Mechanistically, GAS5 functioned as a competing endogenous RNA to inhibit miR-18a-5p and restore the expression of the tumor suppressor CADM2. The direct interactions between GAS5 and miR-18a-5p, and between miR-18a-5p and the CADM2 3′-UTR, were validated by luciferase assays. Rescue assays demonstrated that miR-18a-5p overexpression or CADM2 silencing reversed the tumor-suppressive effects of GAS5. In the xenograft mice, excessive GAS5 significantly impaired tumorigenesis and modulated apoptosis- and EMT-related markers. GAS5 suppresses HCC by modulating the miR-18a-5p-CADM2 axis. Our findings offer a new perspective into HCC tumorigenesis and suggest that GAS5 and its downstream network may serve as potential therapeutic targets.
Camellia drupifera is an economically important woody oil plant rich in edible seed oil. High-quality and well-developed floral buds are key determinants of its yield. However, studies on the regulatory mechanisms underlying its floral bud development remain relatively limited. In this study, proteomics and metabolomics profiles, along with physiological traits, tested during the final three developmental stages of floral buds. The results showed that the contents of indole-3-acetic acid (IAA), abscisic acid (ABA), and 1-aminocyclopropane-1-carboxylic acid (ACC, a precursor of ethylene) decreased significantly in the second (GZII) and third (GZIII) stages. Gibberellin 3 (GA3) maintained a relatively stable content in GZII with a slight decrease, but decreased significantly in GZIII. These changes in hormone contents may promote the growth of stamens and pistils. During the transition among the three floral bud developmental stages, the relative water content, superoxide dismutase (SOD) activity, soluble sugar content, and soluble protein content decreased significantly, accompanied by an increase in hydrogen peroxide (H2O2) content. Integration of differentially accumulated metabolites (DAMs) and differentially expressed proteins (DEPs) analysis showed that the upregulated proteins in the energy metabolism pathway, including pyruvate kinase (PK), citrate synthase (CS), succinate dehydrogenase (SDH), aldolase (ALDO), and hexokinase (HK), promoted the production of adenosine triphosphate (ATP)—a critical energy source for the entire floral development process, including plant hormone synthesis. The decreased IAA content might be attributed to the downregulation of enzymes involved in tryptophan metabolism, such as aldehyde dehydrogenase (ALDH), 3-dehydroquinate dehydratase/shikimate dehydrogenase (aroDE), and chorismate mutase/prephenate dehydratase (aroH). The contents of ABA and GA3 were affected by the downregulation of 4-hydroxy-3-methylbut-2-enyl diphosphate reductase (ispH). Furthermore, the contents of anthocyanins (antioxidant metabolites) decreased significantly in GZII and GZIII, which was caused by the downregulation of key enzymes in the anthocyanin synthesis pathway, including 4-coumarate-CoA ligase (4CL), flavonoid 3’-hydroxylase (F3’H), flavanone 3-hydroxylase (F3H), flavonol synthase (FLS), and anthocyanidin synthase (ANS). This study provides a foundation for further investigating the regulatory metabolites and enzymes involved in C. drupifera floral bud development, and lays a theoretical basis for solving the problem of flower abscission in C. drupifera.
We aimed to investigate whether secretory carrier membrane protein 3 (SCAMP3) knockdown could attenuate hypoxic pulmonary hypertension (HPH) potentially involving WW domain-containing E3 ubiquitin protein ligase 1 (WWP1). Fifty mice were randomly assigned to a blank group, an HPH group, a small interference (si)-negative control (NC) group, a si-SCAMP3 group and a si-SCAMP3 + si-WWP1 group (n = 10 per group). Pulmonary function, right ventricular systolic pressure (RVSP) and the Fulton index [RV/(LV + S)] were measured. Hematoxylin-eosin staining was performed to evaluate pulmonary vascular morphology and the pulmonary vascular remodeling indexes were calculated. Compared with the normoxia group, the hypoxia group exhibited significantly increased SCAMP3 relative expression, OD450 value and SCAMP3 protein expression in HPASMCs, while the apoptosis rate and WWP1 protein expression were significantly decreased (P < 0.05). Compared with the hypoxia group, the hypoxia + si-SCAMP3 group showed significantly reduced SCAMP3 relative expression, OD450 value and SCAMP3 protein expression, but significantly increased apoptosis rate and WWP1 protein expression (P < 0.05). Compared with the hypoxia + si-SCAMP3 group, the hypoxia + si-SCAMP3 + si-WWP1 group exhibited partial restoration of SCAMP3 expression increased OD450 value and SCAMP3 protein expression, and decreased apoptosis rate and WWP1 protein expression (P < 0.05). These findings suggest that SCAMP3 knockdown may inhibit hypoxia-induced proliferation and promote apoptosis potentially involving WWP1, thereby alleviating pulmonary vascular remodeling in HPH mice.
The systematics and sustainable fisheries management of Neolissochilus sumatranus (Weber and de Beaufort 1916) remain challenging due to taxonomic controversies and limited molecular data. Hence, this study utilized next-generation sequencing to generate a mitogenome of N. sumatranus, morphologically identified from northern Sumatra, Indonesia. The assembled mitogenome of this species is 16,584 bp long and consists of 37 genes and a control region (CR). The comparative analysis with nine other Neolissochilus species revealed a highly conserved genetic organization, with relative synonymous codon usage showing high frequencies for arginine, leucine, and serine. Most protein-coding genes (PCGs) in N. sumatranus used ATG as the initiation codon, a pattern widely observed in vertebrate mitogenomes. Conversely, incomplete stop codons (TA- and T–) are proposed to be completed post-transcriptionally through polyadenylation during mRNA maturation. The nonsynonymous-to-synonymous substitution ratios were below ‘1’ for 12 PCGs, except Cytb in Neolissochilus species, indicating that the majority of these genes are subject to purifying selection. The secondary structures of all 22 transfer RNAs in N. sumatranus retained the canonical cloverleaf configuration, displaying no reduction of the dihydrouridine arm typically observed in tRNA-Ser1, thereby reflecting structural variation within cyprinids. The comparative assessment of the CR among Neolissochilus species demonstrated four conserved sequence blocks (CSB-D, CSB-1, CSB-2, and CSB-3) exhibiting species-specific variation, while the tandem repeats were detected in only six of the eight analyzed species. These non-coding regions also exhibited differences in length and nucleotide composition among the examined Neolissochilus species, highlighting their utility for comparative genetic studies. However, their applicability as population-level markers remains to be evaluated using multiple specimens per species to capture intraspecific variation. The mitogenome-based phylogenetic inference corroborates the monophyly of Torinae and suggests a close evolutionary relationship among N. sumatranus, N. hendersoni, and N. soroides, which may be associated with their shared biogeographic history in the Sundaland region. The cladistic relationships between the genera Neolissochilus and Tor remain poorly resolved, with Neolissochilus benasi and Tor remadevii potentially constituting distinct lineages outside the core Neolissochilus and Tor clades, respectively. The phylogenetic positions of Tor douronensis and T. tambra are inconsistent despite their taxonomic synonymy, emphasizing the need for morpho-genetic studies across mainland and island Southeast Asian populations. Overall, the novel mitogenome of N. sumatranus clarifies its phylogenetic position within Torinae and provides a foundation for future systematics study.
Lumbar disc degeneration (LDD) is a major cause of chronic low back pain, and current treatments only provide symptomatic relief. This study aimed to investigate the role of miR-4632-5p and its target MAP3K11 in the pathogenesis of LDD. Bioinformatics analysis was used to predict target genes of miR-4632-5p. LDD tissues (n=22) and normal intervertebral disc (IVD) tissues (n=19) were collected to detect the expression of miR-4632-5p and MAP3K11 by RT-qPCR. A dual-luciferase reporter assay validated their direct interaction. In vitro, TNF-α-induced human nucleus pulposus cells (HNPCs) degeneration models were used to evaluate the effects of miR-4632-5p/MAP3K11 on cell viability, apoptosis, oxidative stress, and inflammatory cytokine secretion via CCK-8, flow cytometry, and ELISA. miR-4632-5p was significantly downregulated in LDD tissues, whereas MAP3K11 was upregulated (P<0.0001), and their expression showed a strong negative correlation (r=-0.82, P<0.0001). miR-4632-5p targeted the 3′UTR of MAP3K11, reducing dual luciferase activity by 35.9
