LncRNA GAS6-AS2 regulates vascular smooth muscle cell senescence through the miR-138-5p/AKT1 axis and serves as a diagnostic and prognostic marker for atherosclerosis.

GAS6-AS2 expression was significantly increased in atherosclerosis patients compared to healthy controls.

• 107 AS patients and 105 healthy controls were included in the study.
• GAS6-AS2 levels were measured using RT-qPCR in peripheral blood samples.
• GAS6-AS2 expression was also elevated in VSMCs treated with ox-LDL, establishing consistency between clinical and cell model findings.
• The increased expression was observed both in vivo (patient samples) and in vitro (ox-LDL-induced cell model).

GAS6-AS2 demonstrated high diagnostic accuracy for atherosclerosis as assessed by ROC curve analysis.

• ROC curve analysis was performed to evaluate the diagnostic value of GAS6-AS2 in AS patients versus healthy controls.
• The study described GAS6-AS2 as showing 'high diagnostic accuracy' for AS.
• GAS6-AS2 also showed risk prediction value for AS patients.
• Kaplan-Meier survival analysis and Cox regression were used to evaluate the prognostic value of GAS6-AS2.

Knockdown of GAS6-AS2 reduced cellular senescence markers in ox-LDL-induced VSMCs.

• SA-β-Gal-positive cells were reduced following GAS6-AS2 knockdown in ox-LDL-treated VSMCs.
• Senescence-related genes and proteins p16, p21, and p53 were downregulated upon GAS6-AS2 knockdown.
• CCK-8 assay was used to assess cell viability/proliferation as part of the biological function assessment.
• SA-β-Gal staining was used as the primary indicator of cellular senescence.

Knockdown of GAS6-AS2 decreased inflammatory cytokine levels in ox-LDL-induced VSMCs.

• Levels of IL-6 and IL-1β were decreased following GAS6-AS2 knockdown.
• Inflammatory factor levels were measured using ELISA.
• The reduction in inflammation occurred in the ox-LDL-induced VSMC atherosclerosis cell model.

GAS6-AS2 directly bound to miR-138-5p and inhibited its expression, while miR-138-5p targeted AKT1 to suppress its expression.

• Bioinformatics prediction was used to identify the regulatory axis.
• Dual-luciferase reporter assay was performed to verify direct binding between GAS6-AS2 and miR-138-5p.
• miR-138-5p was confirmed to target AKT1 and suppress its expression.
• GAS6-AS2 functions as a competing endogenous RNA (ceRNA) by sponging miR-138-5p, thereby upregulating AKT1.

Rescue experiments confirmed that the GAS6-AS2/miR-138-5p/AKT1 axis mediates ox-LDL-induced VSMC senescence and inflammation.

• Rescue experiments were performed to validate the functional axis.
• The axis was confirmed to mediate both senescence and inflammatory responses in ox-LDL-induced VSMCs.
• GAS6-AS2 regulates AKT1 expression indirectly by sponging miR-138-5p, relieving miR-138-5p-mediated suppression of AKT1.
• The complete regulatory mechanism was described as GAS6-AS2 sponging miR-138-5p to upregulate AKT1, promoting VSMC senescence and inflammation.