Ulayatul Kustiati, Dewi Ratih Tirto Sari, Golda Rani Saragih, Chairunisa Isnainingrum, Muhammad Lokman Md Isa, Dwi Aris Agung Nugrahaningsih, Yudy Tjahjono, Dwi Liliek Kusindarta, Srikanth Karnati, Hevi Wihadmadyatami
Lung cancer was among the primary contributors to mortality within the realm of cancer-related ailments. Multiple research projects had demonstrated Ocimum sanctum’s diverse biological and pharmacological characteristics, including its antioxidant, neuroprotective, and anti-cancer capabilities. However, limited evidence supported the assertion that O. sanctum on human lung adenocarcinoma cancer cells. Here, we inspected ethanol extract O. sanctum’s (EEOS) impact on cellular apoptosis and the suppression of Nuclear Factor kappa-light-chain-enhancer of activated B cells (NF-κB) in A549 human lung adenocarcinoma cancer cells. The chemical composition of EEOS was analyzed using gas chromatography-mass spectrometry (GC-MS), while antioxidant potential was assessed via the DPPH radical scavenging assay. Apoptotic activity was evaluated through Hoechst 33342 nuclear staining, mitochondrial membrane potential (JC-1) assay, and Reactive Oxygen Species (ROS) detection using DCFH-DA. Protein expression of NF-κB, Apoptotic Protease Activating Factor 1 (Apaf-1), caspase-9, and caspase-3 was quantified by ELISA. Molecular docking was performed to explore the interactions between linoleic acid-the predominant compound identified in EEOS-and key apoptotic proteins. Additionally, the in vivo chemoprotective effect of EEOS was evaluated in benzo(a)pyrene (B(a)P)-induced lung toxicity in C3H mice. GC-MS profiling identified 33 compounds in EEOS, with linoleic acid, phytol, and β-sitosterol as the major constituents. EEOS exhibited moderate antioxidant activity (IC₅₀ = 46.42 µg/mL). Treatment with EEOS significantly induced apoptosis in A549 cells, disrupted mitochondrial membrane potential, and elevated intracellular ROS levels. ELISA analysis showed downregulation of NF-κB and upregulation of Apaf-1, caspase-9, and caspase-3 in a dose-dependent manner. Molecular docking revealed strong binding affinity of linoleic acid to NF-κB and caspase proteins, comparable to cisplatin. In vivo, EEOS mitigated B[a]P-induced lung tissue damage. In conclusion, EEOS exerts potent pro-apoptotic effects through NF-κB inhibition and activation of the intrinsic Apaf-1/caspase-dependent pathway, supporting its potential as a promising adjunct therapy for lung cancer treatment. © 2026, Walailak University. All rights reserved.
Study Program of Veterinary Science, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Special Region of Yogyakarta, 55281, Indonesia; Laboratory of Veterinary pharmacology, Faculty of Veterinary Medicine, Universitas Brawijaya, East Java, 65151, Indonesia; Department of Pharmacy, Faculty of Medical Science, Universitas Ibrahimy, East Java, 68374, Indonesia; Department of Anatomy and Cell Biology, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Special Region of Yogyakarta, 55281, Indonesia; Department of Basic Medical Science, Kulliyyah of Nursing, International Islamic University Malaysia, Pahang, 53100, Malaysia; Department of Pharmacology and Therapy, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada, Special Region of Yogyakarta, 55281, Indonesia; Biomedical Laboratory, Faculty of Pharmacy, Widya Mandala Catholic University, Surabaya, 60265, Indonesia; Institute of Anatomy and Cell Biology, Julius-Maximillian’s-University Würzburg, Würzburg, 907070, Germany
