PUBLICATION
Identification and Characterization of a Novel Protein ASP-3 Purified from Arca subcrenata and Its Antitumor Mechanism
- Authors
- Guo, Z., Shi, H., Li, C., Luo, Y., Bi, S., Yu, R., Wang, H., Liu, W., Zhu, J., Huang, W., Song, L.
- ID
- ZDB-PUB-190912-2
- Date
- 2019
- Source
- Marine drugs 17(9): (Journal)
- Registered Authors
- Keywords
- Arca subcrenata protein, antitumor mechanism, structural characterization
- MeSH Terms
-
- Angiogenesis Inhibitors/pharmacology
- Animals
- Animals, Genetically Modified
- Antineoplastic Agents/pharmacology*
- Arcidae/chemistry*
- Biological Products/pharmacology*
- Carcinoma, Hepatocellular/drug therapy
- Carcinoma, Hepatocellular/metabolism
- Cell Line
- Cell Line, Tumor
- Cell Proliferation/drug effects
- Female
- Hep G2 Cells
- Human Umbilical Vein Endothelial Cells
- Humans
- Liver Neoplasms/drug therapy
- Liver Neoplasms/metabolism
- Male
- Neovascularization, Pathologic/drug therapy
- Neovascularization, Pathologic/metabolism
- Proteins/pharmacology*
- Signal Transduction/drug effects
- Vascular Endothelial Growth Factor Receptor-2/metabolism
- Xenograft Model Antitumor Assays/methods
- Zebrafish
- PubMed
- 31505835 Full text @ Mar. Drugs
Citation
Guo, Z., Shi, H., Li, C., Luo, Y., Bi, S., Yu, R., Wang, H., Liu, W., Zhu, J., Huang, W., Song, L. (2019) Identification and Characterization of a Novel Protein ASP-3 Purified from Arca subcrenata and Its Antitumor Mechanism. Marine drugs. 17(9):.
Abstract
Diverse bioactive substances derived from marine organisms have been attracting growing attention. Besides small molecules and polypeptides, numerous studies have shown that marine proteins also exhibit antitumor activities. Small anticancer proteins can be expressed in vivo by viral vectors to exert local and long-term anticancer effects. Herein, we purified and characterized a novel protein (ASP-3) with unique antitumor activity from Arca subcrenata Lischke. The ASP-3 contains 179 amino acids with a molecular weight of 20.6 kDa. The spectral characterization of ASP-3 was elucidated using Fourier Transform infrared spectroscopy (FTIR) and Circular Dichroism (CD) spectroscopy. Being identified as a sarcoplasmic calcium-binding protein, ASP-3 exhibited strong inhibitory effects on the proliferation of Human hepatocellular carcinoma (HepG2) cells with an IC50 value of 171.18 ± 18.59 μg/mL, measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The RNA-seq analysis showed that ASP-3 regulated the vascular endothelial growth factor receptor (VEGFR) signaling pathway in HepG2 cells. Immunofluorescence results indicated that ASP-3 effectively reduced VEGFR2 phosphorylation in HepG2 cells and affected the downstream components of VEGF signaling pathways. The surface plasmon resonance (SPR) analysis further demonstrated that ASP-3 direct interacted with VEGFR2. More importantly, the therapeutic potential of ASP-3 as an anti-angiogenesis agent was further confirmed by an in vitro model using VEGF-induced tube formation assay of human umbilical vein endothelial cells (HUVECs), as well as an in vivo model using transgenic zebrafish model. Taken together, the ASP-3 provides a good framework for the development of even more potent anticancer proteins and provides important weapon for cancer treatment using novel approaches such as gene therapy.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping