Gallbladder carcinoma (GBC), the most common malignant tumour of the bile duct, is highly aggressive and has a poor prognosis. MicroRNA-30a-5p (miR-30a-5p) is an important tumour suppressor that participates in many aspects of carcinogenesis and cancer development. However, the role of miR-30a-5p in GBC development remains to be determined, as do the mechanisms underlying its effects in GBC. Using samples collected from 42 subjects with gallbladder carcinoma (GBC), we showed decreased miR-30a-5p expression in the primary lesions vs. non-tumour adjacent tissues (NATs). Decreased miR-30a-5p was associated with shorter disease-free survival (DFS) and overall survival (OS). Inhibiting miR-30a-5p expression in 2 representative GBC cell lines (GBC-SD and NOZ) increased cell proliferation, migration, invasiveness, as well as β-catenin nuclear translocation, vice versa. In nude mice, NOZ cells transfected with miR-30a-5p mimics grew slower (vs. miR-NC) upon subcutaneous inoculation, and had lower rate of hepatic metastasis upon spleen inoculation. Dual luciferase assay confirmed that E2F transcription factor 7 (E2F7) was a direct target of miR-30a-5p and antagonized the effects induced by miR-30a-5p downregulation in GBC cells. MiR-30a-5p attenuates the EMT and metastasis in GBC cells by targeting E2F7, suggesting miR-30a-5p is a tumour suppressor that may serve as a novel potential prognostic biomarker or molecular therapeutic target for GBC.

Background: N⁶-methyladenosine (m⁶A), is the most common modification found in mRNA and lncRNA in higher organisms and plays an important role in physiology and pathology. However, its role in pan-cancer has not been explored. Results: A total of 31 m⁶A modification regulators, including 12 writers, 2 erasers, and 17 readers are identified in the current study. The functional analysis of the regulators results in the enrichment of processes, primarily related to RNA modification and metabolism, and the PPI network reveals multiple interactions among the regulators. The mRNA expression analysis reveals a high expression for most of the regulators in pan-cancer. Most of the m⁶A regulators are found to be mutated across the cancers, with ZC3H13, VIRMA, and PRRC2A having a higher frequency rate. Significant correlations of the regulators with clinicopathological parameters, such as age, gender, tumor stage, and grade are identified in pan-cancer. The m⁶A regulators’ expression is found to have significant positive correlations with the miRNAs in pan-cancer. The expression pattern of the m⁶A regulators is able to classify the tumors into different subclusters as well as into high- and low-risk groups. These tumor groups show differential patterns in terms of their immune cell infiltration, tumor stemness score, genomic heterogeneity score, expression of immune regulatory/checkpoint genes, and correlations between the regulators and the drugs. Conclusions: Our study provide a comprehensive overview of the functional roles, genetic and epigenetic alterations, and prognostic value of the RNA m⁶A regulators in pan-cancer.

Aging and its associated neurodegenerative diseases were increasingly recognized as one of the greatest global health threats. Brain aging involved a multitude of pathological changes, including immune inflammation and metabolic dysregulation. We systematically characterized spatiotemporal heterogeneity and dynamic gene expression profiles related to immune and metabolic dysfunction during mouse brain aging. By analyzing differentially expressed genes and conducting functional enrichment and gene set enrichment analysis, we constructed a spatiotemporal transcriptomic atlas of aging‐associated metabolic and immune features. We further performed transcription factor enrichment analysis, regulatory network construction, weighted gene co-expression network analysis, and single-cell deconvolution analysis to further reveal underlying mechanisms. Our results showed that, throughout aging, most brain regions underwent widespread immune-inflammatory activation with distinct timing and magnitude: svz exhibited the strongest activation at 26 months, th peaks at 21 months, while olf maintained low immune activation. In terms of metabolic function, different regions responded oppositely to oxidative stress and compensatory mechanisms: mPTP pathway’s genes were significantly upregulated in th but downregulated in cor. Moreover, each brain region possessed its own unique regulatory network, and the H3K27-methyltransferase activity of the PRC2 complex displayed region-dependent age dynamics, suggesting these epigenetic differences (especially ent and olf) might serve as novel targets for anti-aging interventions. This work presented a high-resolution spatiotemporal transcriptomic atlas of mouse brain aging, revealed regionally staggered immune activation, contrasted metabolic adaptations, and illustrated PRC2-mediated epigenetic remodeling during brain aging. These insights advanced our understanding of brain aging mechanisms and identified region-specific therapeutic targets for precision interventions in neurodegenerative diseases.