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The growing demand for natural therapeutics with safer pharmacological profiles has renewed interest in aromatic plants traditionally used in folk medicine. Jasonia glutinosa (L.) Fourr. (Asteraceae), commonly known as "sticky fleabane", is a medicinal and aromatic species widely employed for its digestive and anti-inflammatory benefits. Despite its ethnopharmacological relevance, the bioactive potential of its essential oil remains poorly explored. This study provides a multifaceted investigation of the chemical composition, safety profile, and pharmacological activities of J. glutinosa essential oil (JGEO). Gas chromatography-mass spectrometry (GC-MS) analysis revealed nerolidol (51.29%) as the predominant constituent, accompanied by isoaromadendrene epoxide (17.39%) and cis-α-copaene-8-ol (9.92%). The genotoxicity assessment, conducted using the alkaline comet assay on rat leukocytes, demonstrated no significant DNA damage at concentrations up to 100 µg/mL, confirming the oil's genomic safety within the tested range. Pharmacological evaluation showed that JGEO exerted potent anti-inflammatory effects, significantly reducing nitric oxide (NO) and prostaglandin E₂ (PGE₂) production in LPS-stimulated macrophages - with NO levels decreasing to 18.4% of control values and PGE₂ from 720 µM to 108.2 µM (p < 0.001). Moreover, JGEO exhibited selective cytotoxicity against multiple human cancer cell lines, with IC₅₀ values of 15.43 µg/mL (MCF-7), 24.24 µg/mL (MDA-MB-468), 74.57 µg/mL (HCT-15), and 18.06 µg/mL (HepG2). Remarkably, high selectivity indices for MCF-7 (SI = 63.67) and HepG2 (SI = 54.40) surpassed those of doxorubicin, highlighting its favorable therapeutic window. Collectively, these findings provide a scientific basis for the traditional uses of J. glutinosa, emphasizing its safety, efficacy, and chemical richness. The study positions JGEO as a promising natural source of bioactive terpenoids for the development of anti-inflammatory and anticancer agents, supporting its further exploration in pharmaceutical and nutraceutical formulations.