2. Academic Validation
  3. Eurycomanone inhibits osteosarcoma growth and metastasis by suppressing GRP78 expression

Eurycomanone inhibits osteosarcoma growth and metastasis by suppressing GRP78 expression

  • J Ethnopharmacol. 2024 Dec 5:335:118709. doi: 10.1016/j.jep.2024.118709.
Wenyuan Xu 1,  Zhuoying Wang 1,  Tongtong Liu 1,  Xinglong Ma 1,  Ming Jiao 2,  Weisong Zhao 1,  Lingfeng Yu 1,  Yingqi Hua 1,  Zhengdong Cai 3,  Jingjie Li 4,  Tao Zhang 5
Affiliations

Affiliations

  • 1 Department of Orthopedics, Shanghai Bone Tumor Institution, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200080, China.
  • 2 Laboratory Animal Center, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200080, China.
  • 3 Department of Orthopedics, Shanghai Bone Tumor Institution, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200080, China. Electronic address: caizhengdong@sjtu.edu.cn.
  • 4 Precision Research Center for Refractory Diseases, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200080, China. Electronic address: jingjieli@sjtu.edu.cn.
  • 5 Department of Orthopedics, Shanghai Bone Tumor Institution, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200080, China. Electronic address: zhangtaoabc2008@sjtu.edu.cn.
PMID: 39163893 DOI: 10.1016/j.jep.2024.118709
Abstract

Ethnopharmacological relevance: Osteosarcoma (OS) is characterized by rapid growth and frequent pulmonary metastasis. Eurycoma longifolia Jack, a flowering plant primarily found in Southeast Asian countries, is commonly used in traditional herbal medicine. Its root extract is mainly used for against Cancer, malaria, parasites and other conditions. The active compound in its root extract, eurycomanone (EUR), has been proven to inhibit lung and liver Cancer proliferation.

Aim of the study: Our research aimed to investigate the inhibitory effect and underlying molecular mechanism of EUR on OS growth and metastasis.

Materials and methods: In vitro experiments: western blotting (WB) screened 41 compounds that inhibited GRP78 expression and evaluated the protein levels of GRP78, PARP, cleaved-PARP, MMP2, and MMP9. Cell proliferation was evaluated using CCK-8, EdU, colony formation assay, and cell Apoptosis was assessed by flow cytometry. Transwell, wound healing, and tube formation assays were performed to determine the effect of EUR on tumor invasion, migration, and angiogenesis, respectively. Quantitative real-time polymerase chain (qRT-PCR) and dual-luciferase activity assays detected GRP78 mRNA stability and transcription levels post-EUR and thapsigargin treatment. RNA-Seq identified signaling pathways inhibited by EUR. In vivo experiments: effects of EUR in mice were evaluated by H&E staining to detect lung metastasis and potential toxic effects in tissues. Immunohistochemical (IHC) staining detected the expression of Ki-67, CD31, and cleaved Caspase-3 in tumors.

Results: GRP78 is highly expressed in OS and correlated with poor prognosis. In vitro, eurycomanone (EUR) significantly downregulated GRP78 expression, inhibited cell proliferation, migration, invasion, tube formation, and induced Apoptosis. Moreover, it enhanced trichostatin A (TSA) sensitivity and exhibited inhibitory effects on other Cancer types. Mechanistically, EUR decreased GRP78 mRNA stability and transcription. In vivo, EUR inhibited proliferation and invasion in tibial and PDX models.

Conclusions: Our study demonstrated that EUR inhibits the growth and metastasis of OS by reducing GRP78 mRNA stability and inhibiting its transcription, which offers a novel approach for clinical treatment of OS.

Keywords

Eurycomanone; GRP78; Growth; Metastasis; Osteosarcoma.

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