Arsenicum iodatum Induces Apoptosis and Cell Cycle Perturbation in HepG2 Hepatocellular Carcinoma Cells: A Preliminary In Vitro Study

Abstract Hepatocellular carcinoma (HCC) is a leading cause of cancer-related mortality worldwide. Identification of potential therapeutic strategies and molecular targets requires evaluation in robust preclinical models. Several homeopathic preparations have been reported to influence proliferation, apoptosis, and oxidative signaling in cancer cell models. Arsenicum iodatum (Ars. iod), a homeopathic preparation traditionally associated with degenerative and cachectic conditions and clinically used in some HCC patients at the Government Homoeopathic Cancer Centre, Kerala, has not previously been evaluated at the cellular level. The aim of this study was therefore to perform a preliminary investigation of the effects of Ars. iod on the human HCC cell line HepG2 in vitro. HepG2 cells were treated with Ars. iod at three potencies (6C, 12C, and 30C) and compared with untreated control cells. An ethanol-matched vehicle control was used in initial cytotoxicity studies. Cellular responses were assessed using the MTT cell-viability assay, acridine orange/ethidium bromide (AO/EtBr) staining for apoptotic morphology, propidium iodide staining with flow cytometry for cell cycle analysis and indirect ELISA for caspase-3 and caspase-7 protein expression. Experiments were performed in biological triplicate and statistically analyzed. Treatment with Ars. iod was associated with reduced HepG2 cell viability after 24 hours. The calculated IC50 values for the 6C, 12C, and 30C potencies were 59.36, 38.47, and 31.86 μL/mL, respectively, compared with 55.19 μL/mL for the vehicle control. Cell cycle analysis demonstrated redistribution of cells across different phases following treatment, with 6C associated with accumulation in the G2/M phase, 12C with increased S-phase population, and 30C with redistribution across both G0/G1 and G2/M phases. AO/EtBr staining, together with elevated caspase-3 and caspase-7 protein expression levels, confirmed that Ars. iod was capable of inducing apoptosis in HepG2 cells. Under controlled in vitro conditions, homeopathic Ars. iod potencies were associated with potency-dependent cytotoxic, cell cycle perturbation, and apoptotic effects in HepG2 cells. These findings provide preliminary evidence supporting further investigation of the biological effects and underlying mechanisms of Ars. iod in HCC models.