Role and mechanism of palmitic acid-regulated palmitoylation modification of SEC63 protein during endoplasmic reticulum stress in Human Ovarian Granulosa Tumor Cells (KGN)

Objective To investigate the role and mechanism of palmitic acid (PA)-regulated palmitoylation modification of SEC63 protein during endoplasmic reticulum (ER) stress in human ovarian granulosa tumor cells (KGN). Methods Serum samples were collected from 22 patients with polycystic ovary syndrome (PCOS) and 20 control subjects undergoing in vitro fertilization. Metabolomic analysis was performed to quantify serum fatty acid levels. KGN cells were treated with PA (0–400 µM) to assess cytotoxicity, ER stress, oxidative stress (OS), mitochondrial function, and apoptosis. Protective effects of lipoic acid (10 µM) and the ER stress inhibitor 4-phenylbutyric acid (4-PBA, 10 µM) were evaluated. Palmitoylation-modified proteomics and bioinformatics analyses were used to identify key proteins and pathways. A PCOS mouse model was established via dehydroepiandrosterone (DHEA) injection, and PA (100 mg/kg/d) was administered intraperitoneally for 21 days. Ovarian morphology, hormone levels, ER stress, and OS markers were analyzed. Results PA levels were significantly elevated in PCOS patients. In KGN cells, PA induced ER stress (upregulation of CHOP, GRP78, ATF6), OS (increased ROS, MDA, O₂•⁻; decreased antioxidant enzymes), mitochondrial dysfunction (reduced ATP, MMP), and apoptosis. Lipoic acid and 4-PBA ameliorated these effects. Proteomics identified SEC63 as a key palmitoylation-modified protein involved in ER stress. Mutation of SEC63 palmitoylation sites reduced ER stress and OS markers. In PA-treated mice, ovarian morphology showed polycystic characteristics, hormone levels (testosterone, E₂, P₄, LH, FSH) were dysregulated, and ER stress/OS markers were elevated. Conclusion PA induces ER stress and OS in KGN cells and mouse ovarian tissue via palmitoylation modification of SEC63, contributing to PCOS pathogenesis. Lipoic acid and SEC63 modulation may offer therapeutic potential for mitigating PA-induced ovarian dysfunction.