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Cited by 31Cholangiocarcinoma and Gallbladder Cancer StudiesWnt/β-catenin signaling in development and cancerHepatocellular Carcinoma Treatment and Prognosis

Bufalin targeting CAMKK2 inhibits the occurrence and development of intrahepatic cholangiocarcinoma through Wnt/β-catenin signal pathway

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Zhang, Huhu, Dong, Xiaolei, Ding, Xiaoyan, Liu, Guoxiang, Yang, Fanghao, Song, Qinghang, Sun, Hongxiao, Chen, Guang +7 more·2023·Journal of Translational Medicine
DOI 10.1186/s12967-023-04613-6PMC PMC10714474
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Abstract

Background: Intrahepatic cholangiocarcinoma (ICC) accounts for about 15% of primary liver cancer, and the incidence rate has been rising in recent years. Surgical resection is the best treatment for ICC, but the 5-year survival rate is less than 30%. ICC signature genes are crucial for the early diagnosis of ICC, so it is especially important to find its signature genes and therapeutic drug. Here, we studied that bufalin targeting CAMKK2 promotes mitochondrial dysfunction and inhibits the occurrence and metastasis of intrahepatic cholangiocarcinoma through Wnt/β-catenin signal pathway. Methods: IC50 of bufalin in ICC cells was determined by CCK8 and invasive and migratory abilities were verified by wound healing, cell cloning, transwell and Western blot. IF and IHC verified the expression of CAMKK2 between ICC patients and normal subjects. BLI and pull-down demonstrated the binding ability of bufalin and CAMKK2. Bioinformatics predicted whether CAMKK2 was related to the Wnt/β-catenin pathway. SKL2001, an activator of β-catenin, verified whether bufalin acted through this pathway. In vitro and in vivo experiments verified whether overexpression of CAMKK2 affects the proliferative and migratory effects of ICC. Transmission electron microscopy verified mitochondrial integrity. Associated Ca 2+ levels verified the biological effects of ANXA2 on ICC. Results: It was found that bufalin inhibited the proliferation and migration of ICC, and CAMKK2 was highly expressed in ICC, and its high expression was positively correlated with poor prognosis.CAMKK2 is a direct target of bufalin, and is associated with the Wnt/β-catenin signaling pathway, which was dose-dependently decreased after bufalin treatment. In vitro and in vivo experiments verified that CAMKK2 overexpression promoted ICC proliferation and migration, and bufalin reversed this effect. CAMKK2 was associated with Ca 2+ , and changes in Ca 2+ content induced changes in the protein content of ANXA2, which was dose-dependently decreasing in cytoplasmic ANXA2 and dose-dependently increasing in mitochondrial ANXA2 after bufalin treatment. In CAMKK2 overexpressing cells, ANXA2 was knocked down, and we found that reversal of CAMKK2 overexpression-induced enhancement of ICC proliferation and migration after siANXA2. Conclusions: Our results suggest that bufalin targeting CAMKK2 promotes mitochondrial dysfunction and inhibits the proliferation and migration of intrahepatic cholangiocarcinoma through Wnt/β-catenin signal pathway. Thus, bufalin, as a drug, may also be used for cancer therapy in ICC in the future.

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  • Bufalin is a potent anticancer agent used clinically in China to treat various cancers.

    “Bufalin, an active monomer extracted from the Chansu, is a potent anticancer agent used clinically in China to treat various cancers1,2.”

    From Harnessing artificial intelligence to identify Bufalin as a molecular glue degrader of estrogen receptor alpha (2025) · cites this paper as [1]

    The Introduction (page unavailable) calls bufalin a potent anticancer drug, and the Discussion describes antitumor effects across several cancers. But the paper does not say bufalin is used clinically in China to treat cancer; it describes bufalin as a promising drug candidate and reports cell and mouse experiments. The clinical-use portion of the claim is unsupported. (LLM, conf=0.88)

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Authors
Zhang, H., Dong, X., Ding, X., Liu, G., Yang, F., Song, Q., Sun, H., Chen, G., Li, S., Li, Y., Wang, M., Guo, T., Zhang, Z., Li, B., & Yang, L.
Affiliations
  • Qingdao University (CN)
  • Weifang People's Hospital (CN)
Corresponding author
  • Li, Bing
  • Yang, Lina
ORCID
Venue
Journal of Translational Medicine
Publisher
Springer Science and Business Media LLC
Published
2023-12-11
Volume
21
Issue
1
Pages
900-900
ISSN
1479-5876 (ISSN-L)
DOI
10.1186/s12967-023-04613-6
PMC
PMC10714474
OpenAlex
W4389573566
Cited by
31
References
48
Citations by year
2026: 11 · 2025: 13 · 2024: 7
FWCI
8.27
Type
journal article
Language
en
Open access
Licence
Locations
  • Journal of Translational Medicine (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • National Natural Science Foundation of China (No.81803895)
  • Shandong Province Natural Science Foundation (ZR2021YQ57)
  • China Postdoctoral Science Foundation (No.2020M682131, No.2021T140357)
Topics
  • Health Sciences › Medicine › Surgery › Cholangiocarcinoma and Gallbladder Cancer Studies
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Wnt/β-catenin signaling in development and cancer
  • Health Sciences › Medicine › Hepatology › Hepatocellular Carcinoma Treatment and Prognosis
Keywords
bufalin, CAMKK2, Wnt/β-catenin signaling, mitochondrial dysfunction, annexin A2, calcium signaling, tumor cell proliferation, tumor cell migration, metastasis, prognostic factors, anticancer natural products, drug target identification, liver cancer treatment, SKL2001, apoptosis
MeSH terms
Bile Duct Neoplasms, Bile Ducts, Intrahepatic, Bufanolides, Cell Movement, Humans, Gene Expression Regulation, Neoplastic, Cholangiocarcinoma, Mitochondrial Diseases, Cell Line, Tumor, Cell Proliferation, beta Catenin, Calcium-Calmodulin-Dependent Protein Kinase Kinase, Wnt Signaling Pathway

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