Abstract
Acetylcholinesterase (AChE), an enzyme responsible for degradation of acetylcholine, has been identified as a prognostic marker in liver cancer. Although in vivo Ache tumorigenicity assays in mouse are present, no established liver cancer xenograft model in zebrafish using an ache mutant background exists. Herein, we developed an embryonic zebrafish xenograft model using epithelial (Hep3B) and mesenchymal (SKHep1) liver cancer cell lines in wild-type and achesb55 sibling mutant larvae after characterization of cholinesterase expression and activity in cell lines and zebrafish larvae. The comparison of fluorescent signal reflecting tumor size at 3-days post-injection (dpi) revealed an enhanced tumorigenic potential and a reduced migration capacity in cancer cells injected into homozygous achesb55 mutants when compared with the wild-type. Increased tumor load was confirmed using an ALU based tumor DNA quantification method modified for use in genotyped xenotransplanted zebrafish embryos. Confocal microscopy using the Huh7 cells stably expressing GFP helped identify the distribution of tumor cells in larvae. Our results imply that acetylcholine accumulation in the microenvironment directly or indirectly supports tumor growth in liver cancer. Use of this model system for drug screening studies holds potential in discovering new cholinergic targets for treatment of liver cancers.
Highlights
The most commonly occurring liver cancer is hepatocellular carcinoma (HCC), with over half a million new cases emerging yearly and a high death rate[1,2]
We developed an effective xenograft model using liver cancer cells in ache mutant background (Fig. 1)
Larvae were collected at the 3 dpi to extract DNA for ALU DNA quantification and zebrafish ache genotyping (Fig. 1g)
Summary
The most commonly occurring liver cancer is hepatocellular carcinoma (HCC), with over half a million new cases emerging yearly and a high death rate[1,2]. Recombinant AChE treatment decreases cell proliferation in liver cancer cell lines and overexpressing AChE in a nude mice xenograft model decreases tumor size[5]. In light of these findings decreased AChE amount is associated with tumorigenesis and studying its role in tumor microenvironment has potential implications in cancer diagnosis and therapy. ALU based transplant DNA quantification methods have been used to quantify tumor load in mammalian xenotransplantation studies[26,27,28,29] These methods provide high detection power and accuracy even when considerably lower proportion of human cells are found in host (
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.
