Background: Cervical cancer is a severe concern for women throughout the world. This percentage of cancer incidence causes sufferers to die at a high rate. It is believed that the bark of the Pinus merkusii tree contains anti-cancer compounds that inhibit cervical cancer cell growth. Aim: This present study aims to examine the cytotoxic ability of P. merkusii tree bark ethanol extract (PMBE) by inducing apoptosis in HeLa cells. Methods: We administered the PMBE at concentrations of 50, 100, 200, and 400 µg/mL to HeLa cell cultures. We then conducted the MTT cytotoxicity assay, detected apoptosis via Annexin V binding, and observed caspase 9 expression via immunocytochemistry. Results: PMBE showed cytotoxic activity on HeLa cells with an IC50 of 226.6 µg/mL for 24 hours of treatment. PMBE caused early apoptosis in up to 81.31% of HeLa cells, as well as increased caspase-9 expression. Conclusion: Based on this study, PMBE is predicted to have dose-dependent antiproliferative or cytotoxicity effects on the HeLa cell line through the intrinsic pathway apoptosis mechanism. [Open Vet J 2024; 14(10.000): 2628-2633]