Chuining Formula Promotes Apoptosis and Inhibits Glycolysis in Pituitary Tumor by Inhibiting KIF20B

Pituitary tumor, the third most prevalent intracranial tumor, causees severe clinical syndromes that affect the life quality of patients. Chuining formula (CNF) is a traditional Chinese medicine formula used to treat pituitary tumor. This study aimed to determine the major active ingredients of CNF, its effects on pituitary tumor, and the potential underlying mechanisms. The active ingredients of CNF were identified using ultra-high-performance liquid chromatography/Q Exactive Orbitrap Mass. Cell Counting Kit-8 method and flow cytometry assessed cell viability and apoptosis, respectively. Glycolysis was evaluated by detecting glucose uptake, lactic acid production, and LDHA expression. The differentially expressed genes (DEGs) after CNF treatment were screened utilizing high-throughput mRNA sequencing. Chromatin immunoprecipitation assays and luciferase reporter assays were applied to verify the regulatory roles of lactic acid-mediated H3K18 lactylation (H3K18la) and ESR1 in Kif20b transcription. The xenograft tumor was constructed to evaluate tumor growth in vivo. Our analysis identified 71 active ingredients in CNF. CNF reduced tumor cell growth, promoted cell apoptosis, and inhibited glycolysis in vitro and in vivo. Kinesin family member 20B ( Kif20b ) was identified as a downregulated differentially expressed gene. Co-treatment of CNF with Kif20b overexpression or silencing demonstrated that CNF weakened cell growth and glycolysis by repressing Kif20b expression in pituitary tumor. Furthermore, CNF inhibited H3K18la and lactic acid/ESR1-mediated transcription of Kif20b . These findings indicate that CNF reduces tumor cell growth, promotes cell apoptosis, and suppresses glycolysis in vitro and in vivo by inhibiting KIF20B, providing a molecular basis for clinical use of CNF to manage pituitary tumor.