Bufalin inhibits proliferation of non-small cell lung cancer by promoting metabolic reprogramming through inhibition of pyruvate kinase M2

  • Biochem Pharmacol. 2026 Jun 9;251(Pt 2):118150. doi: 10.1016/j.bcp.2026.118150.
Boyu Li  1 ,  Ziwei Han  1 ,  Jinghui Zhang  2 ,  Tiantian Meng  3 ,  Yingshuai Zheng  3 ,  Yuting Guan  4 ,  Liyan Qian  4 ,  Yijuan Chen  4 ,  Xuesong Liu  4 ,  Lutfun Nahar  5 ,  Jiawei Ling  6 ,  Yong Chen  7 ,  Tengfei Xu  8
Affiliations
  • 1. College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China; Jinhua Institute of Zhejiang University, Jinhua 321002, China.
  • 2. Beijing Chaoyang Hospital, Capital Medical University, Beijing 100029, China.
  • 3. Zhejiang University-University of Edinburgh Institute (ZJE), Haining, Zhejiang, China.
  • 4. College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China.
  • 5. Laboratory of Growth Regulators, Palacký University and Institute of Experimental Botany, The Czech Academy of Sciences, Šlechtitelů 27, Olomouc, Czech Republic.
  • 6. College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China. Electronic address: [email protected].
  • 7. College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China; Jinhua Institute of Zhejiang University, Jinhua 321002, China. Electronic address: [email protected].
  • 8. College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China. Electronic address: [email protected].
PMID: 42263882 DOI: 10.1016/j.bcp.2026.118150
Abstract

Non-small cell lung Cancer (NSCLC) is a leading cause of cancer-related mortality worldwide. Bufalin, a bioactive bufadienolide derived from Venenum Bufonis, has shown antitumor activity in multiple Cancer models, but its metabolic effects in NSCLC remain incompletely defined. In this study, we integrated untargeted metabolomics, U-13C6-glucose tracing, Seahorse extracellular flux analysis, and target-engagement assays to characterize bufalin-induced metabolic perturbations in A549 NSCLC cells. Bufalin reduced NSCLC proliferation in vitro and inhibited tumor growth in a xenograft model. Metabolic profiling indicated that bufalin altered glycolytic carbon flow and impaired metabolic flexibility, with concomitant changes in glycolysis-related intermediates and mitochondrial stress. Computational docking, peptide-centric local stability assay (PELSA), cellular thermal shift assay (CETSA), and surface plasmon resonance (SPR) supported a direct interaction between bufalin and Pyruvate Kinase M2 (PKM2), suggesting that PKM2 contributes to bufalin-associated metabolic reprogramming. Functional analysis using PKM2-modulated A549 cells further indicated that PKM2 expression influences cellular tolerance to bufalin-induced oxidative and mitochondrial stress. Because bufalin is a pleiotropic cardiotonic steroid with established targets such as Na+/K + -ATPase, the present findings are interpreted as evidence for a PKM2-involved mechanism rather than an exclusive PKM2-dependent pathway. These results provide a mechanistic basis for further investigation of bufalin as a metabolic modulator in NSCLC.

Keywords
Bufadienolide; Cancer metabolism; Metabolic vulnerability; Mitochondrial stress; Stable isotope tracing; Target engagement.
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