Investigation of the Regulatory Mechanism of Huazhuo Runzao Jiangqi Formula on Esophageal Cancer Utilizing Network Pharmacology, Molecular Docking, and <i>In Vivo</i> and <i>In Vitro</i> Experiments

Background Esophageal cancer, a common digestive malignant neoplasm globally, poses significant health risks. Huazhuo Runzao Jiangqi (HRJ) formula, modified from the classic “Qige Powder” with additions and reductions, has shown clear efficacy in treating esophageal cancer and precancerous lesions in clinical practice and preliminary studies. Objective To explore HRJ’s antiesophageal cancer efficacy and mechanism via component identification, network pharmacology, molecular docking, and in vivo/in vitro tests. Methods UHPLC‐Q‐Orbitrap HRMS analyzed HRJ’s chemical constituents. Network pharmacology identified target proteins and signaling pathways for esophageal cancer, with high‐score proteins selected for molecular docking to confirm possible mechanisms. In vivo , tumor proliferation in nude mice was monitored; hematoxylin–eosin (HE) and immunohistochemistry were used to assess the effect of HRJ on tumor pathological characteristics, proliferation level, and autophagy level. In vitro , the CCK‐8 assay evaluated HRJ’s effect on TE1 cell proliferation; cell migration and invasion assays were performed to observe HRJ’s effect on cell migration and invasion abilities; MDC staining examined autophagosomes; and Western blot and qRT‐PCR analyzed PI3K/Akt/mTOR‐related protein and gene expression. Results It was determined that HRJ contains 46 chemical constituents. Network pharmacology study suggested that the PI3K/Akt/mTOR signaling pathway may be pivotal in the mechanism of HRJ for treating esophageal cancer. In vivo and in vitro studies demonstrated that HRJ exerts an inhibitory effect on the proliferation, migration, and invasion of esophageal cancer. It can enhance the protein and gene expressions of Beclin1, Atg5, and PTEN via modulating the PI3K/Akt/mTOR signaling pathway, therefore promoting autophagy. Conclusion HRJ activates autophagy via the PI3K/Akt/mTOR pathway, modulates key autophagy proteins, and inhibits esophageal cancer progression, providing a novel complementary therapy approach.