Fuzheng Huayu Jiedu formula induces ferroptosis associated with p53/HO-1/GPX4 axis and inhibits malignant progression in A549 lung cancer cells

Abstract

Fuzheng Huayu Jiedu Formula (FHJF) is a traditional Chinese herbal formula with reported anti-tumor properties. However, its mechanism against non-small cell lung cancer (NSCLC) remains unclear. In the present study, we employed human lung adenocarcinoma A549 cells as an in vitro model. Cell proliferation and migration were measured by CCK-8, EdU, and wound-healing assays; apoptosis, cell cycle distribution, ROS, and LPO levels by flow cytometry. Mitochondrial morphology was observed under transmission electron microscopy, and ferroptosis-related gene and protein expression were detected by qPCR and Western blotting. Core therapeutic targets were screened using network pharmacology and proteomics, and ferrostatin-1 (Fer-1) rescue experiments were performed for mechanism validation. The results revealed that FHJF significantly inhibited A549 cell proliferation and migration, induced G2/M phase arrest and apoptosis, increased ROS and LPO levels, decreased mitochondrial membrane potential, and induced typical ferroptosis-associated mitochondrial alterations. Moreover, FHJF upregulated p53 and HO-1 expression while downregulating GPX4, and these effects were reversed by Fer-1. Collectively, these findings indicate that FHJF induces ferroptosis in A549 cells in a manner associated with the p53/HO-1/GPX4 axis, providing a preliminary experimental basis for further mechanistic investigation in this cell line.