Biochemical Genetics最新文献

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.

Colorectal cancer (CRC) liver metastasis remains a major cause of mortality, necessitating novel therapeutic strategies. Hedyotis diffusa Willd -Scutellaria barbata D.Don (HD-SB), a traditional Chinese herbal formulation, has demonstrated anti-tumor potential. However, its mechanism in CRC metastasis, particularly through the regulation of circular RNAs, remains unexplored. A CRC liver metastasis model was established in Balb/c nude mice via intrasplenic injection of HCT-15 cells. Mice received HD-SB treatment. Hematoxylin-eosin staining, Masson staining, and immunohistochemistry (Ki67 and Cytokeratin 19 [CK19]) were carried out to observe pathologic changes in CRC liver metastases. In vitro, HCT-15 cells were treated with tumor necrosis factor-α (TNF-α) to induce metastasis. hsa_circ_0039933 was knocked down and overexpressed in vitro. Transwell, wound healing, EdU, and cell counting kit-8 assays were employed to assess cell invasion, migration and proliferation, respectively. The levels of epithelial-mesenchymal transition (EMT) markers were evaluated by quantitative real-time polymerase chain reaction and western blot. HD-SB treatment dose-dependently reduced hepatic tumor nodules, fibrosis, Ki67 and CK19 expression in vivo, with high-dose HD-SB showing superior efficacy. EMT reverse (upregulated E-cadherin, downregulated N-cadherin/Vimentin) and hsa_circ_0039933 suppression were observed in HD-SB treated CRC mouse. In vitro, HD-SB attenuated TNF-α-induced proliferation, migration, and invasion, synergizing with hsa_circ_0039933 knockdown to suppress EMT and metastatic phenotypes. Conversely, hsa_circ_0039933 overexpression exacerbated TNF-α-driven aggressiveness, which was effectively counteracted by HD-SB. Collectively, HD-SB inhibits CRC liver metastasis by targeting hsa_circ_0039933 and modulating EMT. These findings highlight HD-SB as a promising circRNA-directed therapeutic agent for metastatic CRC.

Gastric cancer (GC) remains one of the most common malignant tumors worldwide, with high incidence and mortality rates. Angiopoietin-like protein 1 (ANGPTL1), a member of the ANGPTL family, is known to function as an anti-angiogenic factor and tumor suppressor. However, its role and underlying mechanisms in GC development have not been investigated and require further investigation. Protein expression levels were analyzed using western blotting and immunofluorescence (IF) assays. Cell viability was assessed using the CCK-8 assay, and cell proliferation was evaluated through colony formation assays. Cell migration and invasion were examined using Transwell assays. Angiogenic capacity was determined through tube formation assays. The VEGFA mRNA expression was detected through RT-qPCR. The level of VEGFA was confirmed through ELISA. The tumor size, volume and weight were confirmed through the in vivo assay. The CD31 protein expression was verified though IHC assay. ANGPTL1 was found to be expressed at lower levels in GC cells, and negatively correlated with VEGFA. ANGPTL1 significantly inhibited GC cell proliferation, migration, and invasion. Furthermore, ANGPTL1 suppressed angiogenesis in vitro. Mechanistically, it was observed that ANGPTL1 overexpression reduced VEGFA expression, and ANGPTL1 can interact with VEGFA. Importantly, reintroduction of VEGFA reversed the inhibitory effects of ANGPTL1 on GC progression. Lastly, VEGFA overexpression retarded the tumor growth in vivo. This study demonstrates that ANGPTL1 inhibits the growth, migration, and angiogenesis of GC cells by downregulating VEGFA expression. These findings suggest that ANGPTL1 may serve as a promising therapeutic target for gastric cancer treatment.

Vascular endothelial dysfunction contributes to obesity-related hypertension, yet its mechanisms remain unclear. This study examined the role of N6-methyladenosine (m6A) in this process using in vivo and in vitro models. C57BL/6J mice fed a high-fat diet (HFD) developed obesity-related hypertension, while oxidized low-density lipoprotein (ox-LDL)-treated human aortic endothelial cells (HAECs) simulated endothelial injury. Global m6A levels were assessed via dot blot, and m6A regulators were quantified by qPCR. Endothelial function was evaluated through viability (CCK-8), apoptosis (flow cytometry), and mitophagy (Western blot and immunofluorescence staining). Mechanistic studies included methylated RNA immunoprecipitation (MeRIP) and dual-luciferase assays. Results showed HFD reduced m6A levels but elevated Fto expression in mice, while ox-LDL decreased m6A levels and upregulated FTO in HAECs. FTO knockdown in HAECs enhanced viability and mitophagy while suppressing apoptosis under ox-LDL stress. Mechanistically, FTO silencing increased m6A methylation on NDRG1 mRNA, leading to its degradation via YTHDF2 recognition. NDRG1 overexpression reversed these effects, restoring apoptosis while reducing mitophagy and viability. These findings reveal that FTO depletion attenuates endothelial dysfunction in obesity-related hypertension by enhancing m6A-mediated suppression of NDRG1, highlighting the FTO/m6A/NDRG1/YTHDF2 axis as a therapeutic target for vascular protection.

Peppermint (Mentha piperita L.) is a valuable medicinal and aromatic herb cultivated globally for its essential oil, prized for its therapeutic and flavouring properties. The productivity and essential oil quality of peppermint is strongly influenced by the intervention of biotic and abiotic elicitors, which eventually induces plant growth dynamics, redox homeostasis, and secondary metabolite biosynthesis. This study evaluates the effect of five plant growth promoting rhizobacteria (PGPRs): Bacillus tequilensis (T1), Bacillus subtilis (T2), Bacterium strain (T3), Bacillus thuringiensis (T4), and Bacillus cereus (T5), to access the modulation efficiency of these plant probionts over morphological, physiological, and biochemical parameters of peppermint variety CIM-Suras. The surface sterilized, uniform sized suckers of a menthol-rich peppermint variety CIM-Suras were inoculated with five aforementioned Bacillus spp., endowed with phosphate solubilization, ammonia production, siderophore production, and Indole Acetic Acid production efficiencies, in triplicate. The results of the study revealed that T2 (Bacillus subtilis) has significantly enhanced the photosynthetic process, contents of chlorophyll (64.71%), accumulation of proline (43.17%), SOD (24.89%), and catalase (56.33%), over the control, followed by T1 (Bacillus tequilensis), and T4 (Bacillus thuringiensis). Treatment with Bacillus subtilis (T2) showing maximum improvement in studied morphological, physiological and biochemical parameters in peppermint suggests the potential of the isolate to abate the oxidative damage of the cells during altering environmental conditions. These findings underscore the efficiency of peppermint-native Bacillus-based PGPRs, especially B. subtilis, for improving crop growth, stress resilience, and essential oil yield in a menthol-rich cultivar. This approach offers a sustainable alternative to synthetic fertilizers to improve peppermint productivity.

Persistent endothelial dysfunction and coagulation abnormalities are increasingly recognized as central components of post-COVID syndrome. However, substantial interindividual variability in long-term coagulation biomarkers suggests the presence of genetically determined susceptibility factors capable of modulating post-infectious thromboinflammatory responses. To determine whether common prothrombotic genetic variants modify long-term D-dimer and INR levels after COVID-19 and to assess their value for precision risk stratification. A controlled observational cohort study included 504 adults aged 18-50 years examined more than two years after PCR-confirmed COVID-19 and 270 control individuals without clinically significant SARS-CoV-2 infection. Genotyping of F2 c.20210G > A, F5 c.1691G > A (Factor V Leiden), and MTHFR c.C677T was performed using real-time PCR. D-dimer and international normalized ratio (INR) were measured monthly for 12 months and averaged. Log-transformed D-dimer was analyzed using multivariable linear regression with interaction terms. All three variants were independent predictors of D-dimer. In adjusted models, F2 GA (+ 31%), F5 GA (+ 37%), MTHFR CT (+ 34%), and MTHFR TT (+ 67%) were associated with higher D-dimer levels (all p < 0.001). Significant amplification of genetic effects was observed in post-COVID individuals (Group × Genotype interaction p < 0.01). INR showed no genotype association. COVID-19 acts as a long-term modifier of inherited thrombophilia phenotypes. Incorporating genetic profiling into post-COVID evaluation may enable precision identification of individuals at risk for persistent hypercoagulability.

Circular RNAs (circRNAs) display cancer-specific expression profiles associated with the prevalence and fatality rates of gastric cancer (GC). Although the functions of certain circRNAs in GC advancement have been clarified, a recently identified circRNA hsa_circ_0002473, require further examination. The current study reveals that hsa_circ_0002473 facilitates gastric cancer progression. Initially, we observed that hsa_circ_0002473 exhibited high expression levels in gastric cancer tissue and cells. Subsequent investigations utilizing the Incucyte S3 imaging system, colony formation assay, scratch wound healing assay, and mouse xenograft models demonstrated that knockdown of hsa_circ_0002473 significantly inhibited the proliferation and migration of GC cells. Additionally, dual-luciferase assays confirmed hsa_circ_0002473 directly binds miR-873-5p and miR-873-5p targeting of the TEAD1 3'UTR. Crucially, rescue experiments demonstrated that miR-873-5p restoration or TEAD1 knockdown reduced hsa_circ_0002473-driven GC cells proliferation and migration. Our findings demonstrate that hsa_circ_0002473 elevates TEAD1 expression by competitively sequestering miR-873-5p, thereby promoting gastric cancer progression.​ This newly identified circRNA regulatory axis could enrich the genetic characteristics of GC, provide a possible medicinal genetic target for GC, and present a new perspective for GC molecular diagnosis.

Emerging evidence has demonstrated the additional therapeutic benefits of ticagrelor in acute coronary syndrome (ACS) patients with diabetes. However, the underlying mechanisms of this association remain elusive. Mendelian randomization (MR) analysis using genome-wide association study (GWAS) data on ticagrelor, plasma proteomics and type 2 diabetes was employed to identify causal mediator proteins. RNA sequencing (RNA-seq) of ticagrelor-treated HepG2 cells revealed the molecular pathways regulating glucose metabolism. Genetically proxied ticagrelor was significantly associated with a reduced risk of diabetes (OR = 0.859, 95% CI: 0.783-0.934, P = 7.98E-05), and 24.41% of this effect was mediated by upregulation of BDH2 protein. In vitro experiments confirmed the enhanced effect of ticagrelor on glucose consumption. Transcriptome analysis revealed that mitochondrial respiratory chain transfer and oxidative phosphorylation (OXPHOS) were significantly enriched, and genes related to ATP biosynthesis were significantly upregulated. These findings highlight the non-platelet function of ticagrelor in maintaining glucose homeostasis, providing insights into potential drug repurposing in the future.

The ACTN3 gene encodes a sarcomeric α-actinin-3 protein, which forms an anti-parallel dimer and constitutes the Z-lines in human skeletal muscle fast-twitch fibers. In human ACTN3, a nonsense mutation that replaces a CGA codon for the 577th arginine (R) residue with a TGA premature termination codon (PTC; specified as X) produces the R577X polymorphism. Since ACTN3 577X mRNA is targeted and degraded by a cellular nonsense-mediated mRNA decay (NMD) system, individuals with the homozygous ACTN3 577XX genotype do not express α-actinin-3 protein in the muscles, resulting in a decrease in speed-oriented athletic performance and muscle mass. The PTC has been a target for translational readthrough using aminoglycoside antibiotics, which enable the full-length α-actinin-3 protein to be produced from the ACTN3 577X gene. However, this effect requires a supraphysiological dose (mM levels) and supportive NMD inhibition. Using expression plasmids and HEK293 cultured cells, in this paper I show that 2,6-diaminopurine (DAP), a recently identified natural compound with translational readthrough activity, can produce a full-length α-actinin-3 protein from the ACTN3 577X gene even when used alone and at a relatively low concentration (µM levels). Most ACTN3 577X alleles likely contain three missense mutations (Q523R, R628C, and R776Q). Full-length α-actinin-3 proteins derived from the ACTN3 577X gene formed more homodimers than α-actinin-3 proteins derived from the ACTN3 577R gene. These results indicate that DAP-induced translational readthrough has the potential to restore function to the lost gene ACTN3 577X in humans.