Abstract
Clonorchis sinensis is a carcinogenic human liver fluke, prolonged infection which provokes chronic inflammation, epithelial hyperplasia, periductal fibrosis, and even cholangiocarcinoma (CCA). These effects are driven by direct physical damage caused by the worms, as well as chemical irritation from their excretory-secretory products (ESPs) in the bile duct and surrounding liver tissues. We investigated the C. sinensis ESP-mediated malignant features of CCA cells (HuCCT1) in a three-dimensional microfluidic culture model that mimics an in vitro tumor microenvironment. This system consisted of a type I collagen extracellular matrix, applied ESPs, GFP-labeled HuCCT1 cells and quiescent biliary ductal plates formed by normal cholangiocytes (H69 cells). HuCCT1 cells were attracted by a gradient of ESPs in a concentration-dependent manner and migrated in the direction of the ESPs. Meanwhile, single cell invasion by HuCCT1 cells increased independently of the direction of the ESP gradient. ESP treatment resulted in elevated secretion of interleukin-6 (IL-6) and transforming growth factor-beta1 (TGF-β1) by H69 cells and a cadherin switch (decrease in E-cadherin/increase in N-cadherin expression) in HuCCT1 cells, indicating an increase in epithelial-mesenchymal transition-like changes by HuCCT1 cells. Our findings suggest that C. sinensis ESPs promote the progression of CCA in a tumor microenvironment via the interaction between normal cholangiocytes and CCA cells. These observations broaden our understanding of the progression of CCA caused by liver fluke infection and suggest a new approach for the development of chemotherapeutic for this infectious cancer.
Highlights
Cholangiocarcinoma (CCA) is an aggressive malignancy of the bile duct epithelia associated with local invasiveness and a high rate of metastases
To assess its contribution to the progression of cholangiocarcinoma (CCA), we developed a 3-dimensional (3D) in vitro culture model that consists of CCA cells (HuCCT1) in direct contact with normal cholangiocytes (H69), which are subsequently exposed to C. sinensis excretory-secretory products (ESPs); this model represents a C. sinensis-associated CCA microenvironment
Proinflammatory cytokines such as IL-6 and TGF-β1 secreted by H69 cells exhibited a crosstalk effect regarding the epithelial-mesenchymal transition of HuCCT1 cells, promoting an increase in the metastatic characteristics of CCA cells
Summary
Cholangiocarcinoma (CCA) is an aggressive malignancy of the bile duct epithelia associated with local invasiveness and a high rate of metastases. The proposed mechanisms of liver fluke-associated cholangiocarcinogenesis include mechanical damage to bile duct epithelia resulting from the feeding activities of the worms, infection-related inflammation, and pathological effects from their excretory-secretory products (ESPs), consisting of a complex mixture of proteins and other metabolites) [4]. These coordinated actions provoke epithelial desquamation, adenomatous hyperplasia, goblet cell metaplasia, periductal fibrosis, and granuloma formation, all contributing to the production of a conducive tumor microenvironment. Malignant cholangiocytes undergo uncontrolled proliferation that leads to the initiation and progression of CCA [5]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
