Anticancer activity of Elsholtzia winitiana var. dongvanensis Phuong on gastric cancer cells: phytochemical profiling, cell proliferation inhibition, senescence induction, and cell cycle arrest

Abstract Background Elsholtzia , a genus within the Lamiaceae family, comprises 42 species predominantly distributed across East and Southeast Asia. Elsholtzia winitiana var. dongvanensis Phuong (EWD) is a subspecies of Elsholtzia winitiana endemic to the Dong Van–Ha Giang Province Mountain region in Vietnam. While the anti-inflammatory and antibacterial activities of Elsholtzia species have been well-documented, data regarding the anticancer potential remain limited. This study investigated the anti-proliferative effects of EWD extract on AGS gastric cancer cells. Results The metabolite profiling of EWD leaf extract conducted using ultra-performance liquid chromatography, coupled with quadrupole time-of-flight mass spectrometry identified 65 compounds, including triterpenoids, sesquiterpenoids, alkaloids, glycosides, and phenolic compounds. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay showed that the extract significantly inhibited AGS cell proliferation in a concentration-dependent manner. The cell proliferation rate decreased from 64.7 ± 6.5% to 18.9 ± 1.3% as concentrations increased from 50 to 500 µg/mL, with an IC 50 value of 68.7 µg/mL. In the 3D culture model, the EWD extract inhibited tumorsphere formation, resulting in a significant reduction in tumorsphere numbers compared to controls (100%). Additionally, EWD extract induced cellular senescence, with a 31-fold increase in senescent cells at 50 µg/mL and an up to 82-fold increase at 200 µg/mL. Cell cycle analysis indicated that the EWD extract triggered G0/G1 phase arrest, as evidenced by a significant increase in cell proportion at the G0/G1 phase, 59.7% at 100 µg/mL, and 64.6% at 200 µg/mL, compared to 55.9% in the control. Gene expression analysis revealed significant downregulation of cyclin B1, cyclin D2, cyclin E1, cyclin-dependent kinase 2, and cyclin-dependent kinase 3, along with increased cyclin-dependent kinase 4 expression. Conclusion The EWD extract significantly inhibited proliferation, induced senescence, arrested the cell cycle at the G0/G1 phase, and downregulated cyclin and cyclin-dependent kinase genes in the AGS cells, highlighting its potential for gastric cancer therapy.