Betulinic acid induces lysosome-dependent death in prostate cancer by targeting DDX5

  • Phytomedicine. 2026 Jun:155:158143. doi: 10.1016/j.phymed.2026.158143.
Shoushi Liu  1 ,  Xiaolan Li  1 ,  Zhiping Cheng  1 ,  Kaili Zhao  1 ,  Yanting Wang  1 ,  Yuhe Wei  1 ,  Yuanshuo Wang  1 ,  Jinjian Lu  2 ,  Xin Yang  3 ,  Yang Mooi Lim  4 ,  Qinpei Lu  5 ,  Qiuju Huang  6 ,  Hongwei Guo  7
Affiliations
  • 1. Key Laboratory of Longevity and Aging-related Diseases of Chinese Ministry of Education& Center for Translational Medicine, Nanning 530021, China; Guangxi Key Laboratory of Bioactive Molecules Research and Evaluation & College of Pharmacy, Guangxi Medical University, Nanning 530021, China.
  • 2. State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Macao SAR 999078, China.
  • 3. Key Laboratory of Longevity and Aging-related Diseases of Chinese Ministry of Education& Center for Translational Medicine, Nanning 530021, China.
  • 4. M. Kandiah Faculty of Medicine and Health Sciences, Universiti Tunku Abdul Rahman, Bandar Sungai Long, 43000, Kajang Selangor, Malaysia.
  • 5. Guangxi Key Laboratory of Bioactive Molecules Research and Evaluation & College of Pharmacy, Guangxi Medical University, Nanning 530021, China. Electronic address: [email protected].
  • 6. Guangxi Key Laboratory of Bioactive Molecules Research and Evaluation & College of Pharmacy, Guangxi Medical University, Nanning 530021, China. Electronic address: [email protected].
  • 7. Key Laboratory of Longevity and Aging-related Diseases of Chinese Ministry of Education& Center for Translational Medicine, Nanning 530021, China; Guangxi Key Laboratory of Bioactive Molecules Research and Evaluation & College of Pharmacy, Guangxi Medical University, Nanning 530021, China. Electronic address: [email protected].
PMID: 41967235 DOI: 10.1016/j.phymed.2026.158143
Abstract

Background: Prostate Cancer (PCa) represents a leading cause of cancer-related mortality in men. Betulinic acid (BA) exhibits antitumor properties. However, its direct molecular target and mechanism in PCa remain unclear.

Purpose: To identify BA's molecular target and elucidate its mechanism in inhibiting PCa progression.

Study design: In vitro and in vivo models were employed to evaluate BA's antitumor effects and underlying mechanisms.

Methods: BA's antitumor activity was assessed in PCa cell lines and xenograft models. Apoptosis was examined via Hoechst staining, flow cytometry, and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay. Lysosomal membrane permeabilization (LMP) was examined via LysoTracker Red and acridine orange (AO) staining, validated by CA074-Me. Target identification utilized pull-down assays, drug affinity responsive target stability (DARTS), molecular docking, and microscale thermophoresis (MST). Clinical data analysis and siRNA knockdown confirmed DDX5's role.

Results: BA significantly suppressed PCa growth in vitro and in vivo without toxicity. BA directly bound DDX5, inhibiting transcription factor EB (TFEB)-mediated lysosomal biogenesis and downregulating V-ATPase subunit H (ATP6V1H), essential for lysosomal acidification. This disruption induced LMP and lysosome-dependent cell death (LDCD) via Apoptosis, confirmed by CA074-Me. DDX5 expression was decreased in PCa tissues. DDX5 knockdown abolished BA-induced LDCD and lysosomal dysfunction.

Conclusion: BA directly targets DDX5, inducing LDCD through the novel BA-DDX5-TFEB-ATP6V1H axis, exploiting lysosomal vulnerability in PCa. These findings underscore BA's therapeutic potential and propose lysosomal membrane destabilization as a precision treatment strategy for PCa.

Keywords
Betulinic acid; DEAD-box helicase 5 (DDX5); Lysosomal membrane permeabilization (LMP); Lysosome-dependent cell death (LDCD); Prostate cancer (PCa).
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