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Published on: June 26, 2018
Antitumor Activity of Ruditapes philippinarum Polysaccharides Through Mitochondrial Apoptosis in Cellular and
Mengyue Liu1, Weixia Wang2, Haoran Wang3
1College of Life Sciences, Qingdao University, Qingdao 266071, China.
Abstract:
Colorectal cancer (CRC) remains a predominant cause of global cancer-related mortality, highlighting the pressing demand for innovative therapeutic strategies. Natural polysaccharides have emerged as promising candidates in cancer research due to their multifaceted anticancer mechanisms and tumor-suppressive potential across diverse malignancies. In this study, we enzymatically extracted a polysaccharide, named ERPP, from Ruditapes philippinarum and comprehensively evaluated its anti-colorectal cancer activity. We conducted in vitro assays, including CCK-8 proliferation, clonogenic survival, scratch wound healing, and Annexin V-FITC/PI apoptosis staining, and the results demonstrated that ERPP significantly inhibited HT-29 cell proliferation, suppressed colony formation, impaired migratory capacity, and induced apoptosis. JC-1 fluorescence assays provided further evidence of mitochondrial membrane potential (MMP) depolarization, as manifested by a substantial reduction in the red/green fluorescence ratio (from 10.87 to 0.35). These antitumor effects were further validated in vivo using a zebrafish HT-29 xenograft model. Furthermore, ERPP treatment significantly attenuated tumor angiogenesis and downregulated the expression of the vascular endothelial growth factor A (Vegfaa) gene in the zebrafish xenograft model. Mechanistic investigations revealed that ERPP primarily activated the mitochondrial apoptosis pathway. RT-qPCR analysis showed an upregulation of the pro-apoptotic gene Bax and a downregulation of the anti-apoptotic gene Bcl-2, leading to cytochrome c (CYCS) release and caspase-3 (CASP-3) activation. Additionally, ERPP exhibited potent antioxidant capacity, achieving an 80.2% hydroxyl radical scavenging rate at 4 mg/mL. ERPP also decreased reactive oxygen species (ROS) levels within the tumor cells, thereby augmenting anticancer efficacy through its antioxidant activity. Collectively, these findings provide mechanistic insights into the properties of ERPP, underscoring its potential as a functional food component or adjuvant therapy for colorectal cancer management.
Insights
A novel polysaccharide, ERPP, from Ruditapes philippinarum shows significant anti-colorectal cancer (CRC) activity by inducing apoptosis and inhibiting tumor growth. This natural compound also possesses antioxidant properties, highlighting its potential for CRC management.
Area of Science:
- Marine Biotechnology
- Cancer Research
- Natural Product Chemistry
Background:
- Colorectal cancer (CRC) is a leading cause of cancer mortality globally, necessitating novel therapeutic approaches.
- Natural polysaccharides are recognized for their diverse anticancer mechanisms and tumor-suppressive potential.
- Ruditapes philippinarum, a marine resource, offers a potential source for bioactive compounds.
Purpose of the Study:
- To isolate and characterize a polysaccharide (ERPP) from Ruditapes philippinarum.
- To evaluate the in vitro and in vivo anti-colorectal cancer (CRC) efficacy of ERPP.
- To elucidate the underlying mechanisms of ERPP's anticancer activity.
Main Methods:
- Enzymatic extraction of ERPP from Ruditapes philippinarum.
- In vitro assays: CCK-8 proliferation, clonogenic survival, scratch wound healing, Annexin V-FITC/PI apoptosis, JC-1 fluorescence for mitochondrial membrane potential (MMP).
- In vivo studies using a zebrafish HT-29 xenograft model, including angiogenesis assessment and gene expression analysis (Vegfaa, Bax, Bcl-2, CYCS, CASP-3) via RT-qPCR.
- Antioxidant capacity evaluation (hydroxyl radical scavenging, ROS levels).
Main Results:
- ERPP significantly inhibited HT-29 cell proliferation, colony formation, and migration while inducing apoptosis in vitro.
- ERPP treatment led to MMP depolarization, evidenced by a reduced red/green fluorescence ratio.
- In vivo, ERPP attenuated tumor angiogenesis and downregulated Vegfaa expression in the zebrafish model.
- ERPP activated the mitochondrial apoptosis pathway, increasing Bax/Bcl-2 ratio, promoting cytochrome c release, and activating caspase-3.
- ERPP demonstrated potent antioxidant activity, scavenging hydroxyl radicals and reducing intracellular ROS levels.
Conclusions:
- ERPP exhibits significant anti-CRC properties through the induction of mitochondrial apoptosis and antioxidant mechanisms.
- ERPP effectively suppresses tumor growth, angiogenesis, and metastasis.
- ERPP shows promise as a functional food component or adjuvant therapy for colorectal cancer management.
