2. Academic Validation
  3. Microcystin-leucine arginine induces the proliferation of cholangiocytes and cholangiocarcinoma cells through the activation of the Wnt/β-catenin signaling pathway

Microcystin-leucine arginine induces the proliferation of cholangiocytes and cholangiocarcinoma cells through the activation of the Wnt/β-catenin signaling pathway

  • Heliyon. 2024 Apr 26;10(9):e30104. doi: 10.1016/j.heliyon.2024.e30104.
Suppakrit Kongsintaweesuk 1 2 3,  Sirinapha Klungsaeng 4 3,  Kitti Intuyod 5 3,  Anchalee Techasen 1 6 3,  Chawalit Pairojkul 5 3,  Vor Luvira 7 3,  Somchai Pinlaor 4 3,  Porntip Pinlaor 1 6 3
Affiliations

Affiliations

  • 1 Centre for Research and Development of Medical Diagnostic Laboratories, Faculty of Associated Medical Sciences, Khon Kaen University, Khon Kaen 40002, Thailand.
  • 2 Medical Sciences Program, Faculty of Associated Medical Sciences, Khon Kaen University, Khon Kaen 40002, Thailand.
  • 3 Cholangiocarcinoma Research Institute, Khon Kaen University, Khon Kaen 40002, Thailand.
  • 4 Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand.
  • 5 Department of Pathology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand.
  • 6 School of Medical Technology, Faculty of Associated Medical Sciences, Khon Kaen University, Khon Kaen 40002, Thailand.
  • 7 Department of Surgery, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand.
PMID: 38720699 DOI: 10.1016/j.heliyon.2024.e30104
Abstract

Background: Microcystin-leucine arginine (MC-LR) is a cyanobacterial hepatotoxic toxin found in water sources worldwide, including in northeastern Thailand, where opisthorchiasis-associated cholangiocarcinoma (CCA) is most prevalent. MC-LR is a potential carcinogen; however, its involvement in liver fluke-associated CCA remains ambiguous. Here, we aimed to evaluate the effect of MC-LR on the progression of CCA via the Wnt/β-catenin pathway in vitro.

Methods: Cell division, migration, cell cycle transition, and MC-LR transporter expression were evaluated in vitro through MTT assay, wound healing assay, flow cytometry, and immunofluorescence staining, respectively. Following a 24-h treatment of cultured cells with 1, 10, 100, and 1,000 nM of MC-LR, the proliferative effect of MC-LR on the Wnt/β-catenin signaling pathway was investigated using immunoblotting and qRT-PCR analysis. Immunohistochemistry was used to determine β-catenin expression in CCA tissue compared to adjacent tissue.

Results: Human immortalized cholangiocyte cells (MMNK-1) and a human cell line established from opisthorchiasis-associated CCA (KKU-213B) expressed the MC-LR transporter and internalized MC-LR. Exposure to 10 nM and 100 nM of MC-LR notably enhanced cells division and migration in both cell lines (P < 0.05) and markedly elevated the percentage of S phase cells (P < 0.05). MC-LR elevated PP2A expression by activating the Wnt/β-catenin signaling pathway and suppressing Phosphatase activity. Inhibition of the β-catenin destruction complex genes (Axin1 and APC) led to the upregulation of β-catenin and its downstream target genes (Cyclin D1 and c-Jun). Inhibition of Wnt/β-catenin signaling by MSAB confirmed these results. Additionally, β-catenin was significantly expressed in cancerous tissue compared to adjacent areas (P < 0.001).

Conclusions: Our findings suggest that MC-LR promotes cell proliferation and progression of CCA through Wnt/β-catenin pathway. Further evaluation using invivo experiments is needed to confirm this observation. This finding could promote health awareness regarding MC-LR intake and risk of CCA.

Keywords

Bile duct cancer; Carcinogenesis; Cyanobacteria; Cyanotoxin; Environmental exposure.

Figures
Products
  • Cat. No.
    Product Name
    Description
    Target
    Research Area
  • HY-120697
    99.77%, Wnt/β-Catenin Inhibitor