Genistein (GEN) exhibits concentration- and cell-type-dependent biological activities; however, the mechanisms underlying its cytotoxicity remain incompletely understood. This study investigated whether oxidative stress-mediated mitochondrial dysfunction contributes to GEN-induced apoptosis in TM4 mouse Sertoli cells and ELT3 rat leiomyoma cells. Cells were exposed to GEN (30 or 100 µM) for 48 h. Cytotoxicity, oxidative stress, mitochondrial function, and apoptosis were assessed by measuring cell viability, lactate dehydrogenase release, reactive oxygen species (ROS), malondialdehyde, glutathione reductase activity, mitochondrial membrane potential (ΔΨm), intracellular ATP, apoptosis-related gene expression, and caspase-3/-9 activities. The involvement of oxidative stress was examined using N-acetyl-L-cysteine (NAC). GEN reduced cell viability, antioxidant capacity, ΔΨm, and ATP content while increasing membrane damage, ROS accumulation, and lipid peroxidation in both cell lines. These changes were accompanied by upregulation of Bax, Tp53, caspase-3, and caspase-9; downregulation of Bcl-2; and increased caspase-3/-9 activities. NAC pretreatment attenuated these alterations, supporting ROS as an upstream mediator. Notably, TM4 cells were approximately 4.4-fold more sensitive to GEN than ELT3 cells at 48 h. Collectively, the findings support a ROS-dependent mitochondrial apoptotic response under elevated in vitro GEN exposure but do not establish selective antitumor cytotoxicity or direct physiological relevance.
Samak Sutjarit, C. Sakulthaew, Nattakan Meekhanon et al.· Journal of Xenobiotics· 0 citations
Uterine leiomyoma is the most common benign gynecological tumor and a major cause of abnormal uterine bleeding, pelvic pain, and infertility. Although apigenin (API) exhibits antioxidant, antiproliferative, and anticancer activities, its mechanism of action in uterine leiomyoma, particularly the involvement of oxidative stress-mediated mitochondrial apoptosis, remains unclear. This study investigated the role of oxidative stress in API-induced apoptosis using rat uterine leiomyoma ELT3 cells. Cells were treated with 50 and 100 µM API for 48 h. Oxidative stress was assessed by measuring reactive oxygen species (ROS), malondialdehyde (MDA), and glutathione reductase (GR) activity. Mitochondrial membrane potential (ΔΨm), cell proliferation, lactate dehydrogenase (LDH) release, and apoptosis-related gene and protein expression were also evaluated. API significantly increased ROS and MDA levels while decreasing GR activity, indicating oxidative stress. These changes were accompanied by ΔΨm collapse, reduced cell proliferation, increased LDH release, upregulation of p53, Bax, caspase-3, and caspase-9, and downregulation of Bcl-2, indicating activation of the intrinsic mitochondrial apoptotic pathway. These findings demonstrate that API induces intrinsic mitochondrial apoptosis through oxidative stress-mediated mechanisms, providing mechanistic evidence for its cytotoxic effects and supporting its potential as a phytochemical candidate for uterine leiomyoma treatment.
Samak Sutjarit, C. Setthawongsin, C. Sakulthaew et al.· Toxicology Reports· 0 citations
