Identification and Characterization of a Novel Protein ASP-3 Purified from Arca subcrenata and Its Antitumor Mechanism

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.

Developing natural marine products for treating liver diseases

“…Some data have shown that polypeptides derived from cockles can exert anti-tumor activity by inhibiting the MAPK signaling pathway[ 55 ]. Guo et al [ 56 ] found that the ASP-3 protein isolated from Arca subcrenata inhibited HepG2 cell proliferation.…”

Non-coding RNA-related antitumor mechanisms of marine-derived agents

“…ASP-3 is a protein purified from marine Arca subcrenata and exhibits a strong inhibitory effect on the proliferation of HCC cells ( Guo et al, 2019 ). Mechanistically, ASP-3 was found to effectively reduce vascular endothelial growth factor receptor (VEGFR) 2 phosphorylation (p-VEGFR2) in tumor cells and affect the vascular endothelial growth factor (VEGF) signaling pathway.…”

Marine Natural Products: A Potential Source of Anti-hepatocellular Carcinoma Drugs

“…Recently, the anti-cancer properties of A. subcrenata -derived active materials have been reported in HeLa and HT-29 cells. , Specifically, the protein ASP-3 (MW = 14774.7), isolated from A. subcrenata Lischke, exerts anti-proliferative activity in HepG2 cells, with an IC 50 of 171.18 ± 18.59 μg/mL (IC 50 = 11.59 ± 1.25 μM). Moreover, in vitro studies suggest that it may exert its anti-tumor effects by inhibiting the VEGF signaling pathway …”

“…JG3 (51) and JG6 (52) , two structurally novel marine-derived oligosaccharides, have been shown to possess anti-angiogenic and anti-metastatic effects , as well as anti-cancer effects against liver cancer in vivo and in vitro . − Specifically, JG6 suppresses BEL-7402 cell proliferation and migration in a concentration-dependent manner by inhibiting the C-X-C motif chemokine ligand 12/C-X-C motif chemokine receptor 4 axis. Additionally, the proliferation rates of BEL-7402 cells were inhibited by 22.2%, 46.5%, and 51.0% following treatment with 10, 50, and 100 μg/mL JG6, respectively.…”

“…Moreover, in vitro studies suggest that it may exert its antitumor effects by inhibiting the VEGF signaling pathway. 114 Since 1968, when bryostatin 1 (50) was first isolated from its marine source organism Bugula neritina, several methods have been developed for its artificial synthesis. 115−117 Bryostatin 1 (50) has demonstrated activity against human immunodeficiency virus, 116 leukemia, 119 and Alzheimer's disease.…”