Anticancer Activity of Fucoidan via Apoptosis and Cell Cycle Arrest on Cholangiocarcinoma Cell.

Pathanin Chantree,Pongsakorn Martviset,Kesara Na-Bangchang

doi:10.31557/apjcp.2021.22.1.209

Pathanin Chantree, Pongsakorn Martviset + Show 1 more

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https://doi.org/10.31557/apjcp.2021.22.1.209

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Abstract

Objective:Many previous studies reported that fucoidan has antitumor activities. The objective of the present study was to determine the cytotoxic effects and related mechanisms of cell death induced by fucoidan extracted from Fucus vesiculosus on CL-6 cholangiocarcinoma cell. Methods:CL-6 and OUMS cells were treated with 0, 100, 200, and 300 μg/mL of fucoidan. MTT assay was used to determine cytotoxicity. Flow cytometry-based assay was used to examine the distribution of apoptosis and cell cycle. The changes in nuclear morphology were determined using Hoechst 33,342 staining. Mitochondrial membrane potential (ΔΨm) was evaluated using the JC-1 kit. The apoptotic, anti-apoptotic, and cell cycle-related proteins study were examined by Western blot analysis. Results:The relative viable cell number of treated CL-6 cells was decreased but no effect was observed in OUMS normal cells. Furthermore, treated cells were arrested in the G0/G1 phase with down-regulation of cyclin D1 and CDK4. Annexin V/PI staining with flow cytometry analysis suggested that fucoidan could induce apoptosis in CL-6 cells. Western blot study revealed the up-regulation of apoptotic markers including Bax, cleaved PARP, cleaved caspase-3, but down-regulation of anti-apoptotic markers, cl-2. Moreover, fucoidan could induce nuclear fragmentation and chromatin condensation with alteration of ΔΨm. Conclusion:Fucoidan exerts antitumor properties against CL-6 cholangiocarcinoma cells illustrated by the induction of apoptosis and cell cycle arrest.

Highlights

Cholangiocarcinoma (CCA) is a highly offensive malignancy arising from the epithelium of the biliary tract which consists of intrahepatic and extrahepatic cholangiocarcinoma (Valle et al, 2016)
Cytotoxicity of fucoidan on CL-6 and OUMS cells After treatment for 24 h with fucoidan, cell viability percents of CL-6 cells were decreased in a concentrationdependent manner from 50 to 400 μg/mL compared with the untreated group (Figure 1A)
According to IC50, CL-6 and OUMS cells were subsequently treated with fucoidan (0, 100, 200, and 300 μg/mL) for 24, 48, and 72 h

Summary

Introduction

Cholangiocarcinoma (CCA) is a highly offensive malignancy arising from the epithelium of the biliary tract which consists of intrahepatic (iCCA) and extrahepatic cholangiocarcinoma (eCCA) (Valle et al, 2016). The cell surface receptor tyrosine kinase c-Met only presents in progenitor and stem cells involved in organogenesis and wound healing This receptor is anomalously high in cholangiocarcinoma along with its ligand hepatocyte growth factor (HGF) leading to enhance cell proliferation, angiogenesis, and metastasis (Socoteanu et al, 2008; Leelawat et al, 2006). A key molecule in angiogenesis, vascular endothelial growth factor (VEGF), is highly expressed in cholangiocarcinoma both in the tissue samples and cell lines (Ogasawara et al, 2001) This molecular characteristic is facilitated by the presentation of the estrogen receptors that respond to 17-β estradiol resulting in increased VEGF production and angiogenesis (Alvaro et al, 2006; Mancino et al, 2009). A follow-up study with an international multicenter cohort of patients found a 5-year survival of 65% in patients with very early CCA (Sapisochin et al, 2016)

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