PUBLICATION
1β-OH-arenobufagin induces mitochondrial apoptosis in hepatocellular carcinoma through the suppression of mTOR signaling pathway
- Authors
- Deng, L.J., Lei, Y.H., Quan, J.Y., Li, B.J., Zhang, D.M., Tian, H.Y., Chen, Y., Zhang, E.X., Chen, L., Ye, W.C., Ning, W.M., Yu, L.Z., Liu, J.S.
- ID
- ZDB-PUB-201007-11
- Date
- 2020
- Source
- Journal of ethnopharmacology 266: 113443 (Journal)
- Registered Authors
- Keywords
- 1β-OH-arenobufagin, liver cancer, mTOR, mitochondria apoptosis, zebrafish
- MeSH Terms
-
- Animals
- Antineoplastic Agents/chemistry
- Antineoplastic Agents/isolation & purification
- Antineoplastic Agents/pharmacology*
- Apoptosis/drug effects
- Bufanolides/chemistry
- Bufanolides/isolation & purification
- Bufanolides/pharmacology*
- Carcinoma, Hepatocellular/drug therapy*
- Carcinoma, Hepatocellular/pathology
- Hep G2 Cells
- Hepatocytes/drug effects
- Hepatocytes/metabolism
- Humans
- Liver Neoplasms/drug therapy*
- Liver Neoplasms/pathology
- Mitochondria/drug effects
- TOR Serine-Threonine Kinases/metabolism
- Xenograft Model Antitumor Assays
- Zebrafish
- PubMed
- 33022344 Full text @ J. Ethnopharmacol.
Citation
Deng, L.J., Lei, Y.H., Quan, J.Y., Li, B.J., Zhang, D.M., Tian, H.Y., Chen, Y., Zhang, E.X., Chen, L., Ye, W.C., Ning, W.M., Yu, L.Z., Liu, J.S. (2020) 1β-OH-arenobufagin induces mitochondrial apoptosis in hepatocellular carcinoma through the suppression of mTOR signaling pathway. Journal of ethnopharmacology. 266:113443.
Abstract
Ethnopharmacological relevance Chansu, dried secretions from Bufonidae, has long been used for cancer treatment as a traditional Chinese medicine. In searching for effective anti-hepatoma agents from Chansu, our preliminary drug screening found that a bufadienolide, namely 1β-hydroxyl-arenobufagin (1β-OH-ABF), displays anti-hepatoma activities. However, the anti-hepatoma effects and molecular mechanisms of 1β-OH-ABF have not been defined.
Aim of the study To evaluate the anti-hepatoma activity of 1β-OH-ABF against liver cancer Hep3B and HepG2 cells in vitro and in vivo, as well as explore the underlying mechanisms.
Materials and methods The anti-proliferative effects of 1β-OH-ABF on liver cancer Hep3B, HepG2, HuH7, SK-HEP-1 and normal hepatocyte LO2 cells were examined by MTT assay and colony formation assay. Hoechst 33258 staining and Annexin V-FITC/PI staining assay were used to analyze apoptosis induced by 1β-OH-ABF. The collapse of the mitochondrial membrane potential (ΔΨm) was detected by JC-1 staining assay. Western blotting was used to examine the expression levels of targeted proteins. The role of mTOR in 1β-OH-ABF-induced apoptosis was investigated using small interfering RNA (siRNA) transfection. Zebrafish xenograft model was established to evaluate the anti-hepatoma effects of 1β-OH-ABF in vivo.
Results We found that 1β-OH-ABF inhibits the proliferation of Hep3B, HepG2, HuH7, SK-HEP-1 cells but has little cytotoxicity towards LO2 cells. 1β-OH-ABF induces mitochondria dysfunction and triggers mitochondria apoptotic pathway, which is accompanied by the loss of ΔΨm, upregulation and translocation of Bax, as well as cleavages of caspase-9, caspase-3 and PARP. Mechanistically, 1β-OH-ABF markedly decreases the expression level of p-AKT/AKT and p-mTOR (Ser2248 and Ser2481)/mTOR in a time-dependent manner. Inhibition of mTOR by siRNA strengthens 1β-OH-ABF-mediated apoptosis. Critically, 1β-OH-ABF shows a marked in vivo anti-hepatoma effect on human Hep3B cell xenografts in zebrafish model.
Conclusion 1β-OH-ABF induces mitochondrial apoptosis through the suppression of mTOR signaling in vitro and in vivo, indicating that 1β-OH-ABF may serve as a potential agent for the treatment of liver cancer.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping