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Functional and genomic characterization of three novel cell lines derived from a metastatic gallbladder cancer tumor

Journal article
Authors Patricia García
Carolina Bizama
Lorena Rosa
Jaime A. Espinoza
Helga Weber
Javier Cerda-Infante
Marianela Sánchez
Viviana P. Montecinos
Justo Lorenzo-Bermejo
Felix Boekstegers
Marcela Davila Lopez
Francisca Alfaro
Claudia Leiva-Acevedo
Zasha Parra
Diego Romero
Sumie Kato
Pamela Leal
Marcela Lagos
Juan Carlos Roa
Published in Biological Research
Volume 53
ISSN 07169760
Publication year 2020
Published at Core Facilities, Bioinformatics
Language en
Links https://doi.org/10.1186/s40659-020-...
Keywords Ascites, Cancer cell lines, Gallbladder cancer, Gene expression profile, Native American ancestry
Subject categories Clinical Medicine

Abstract

© 2020 The Author(s). Background: Gallbladder cancer (GBC) is the most common tumor of the biliary tract. The incidence of GBC shows a large geographic variability, being particularly frequent in Native American populations. In Chile, GBC represents the second cause of cancer-related death among women. We describe here the establishment of three novel cell lines derived from the ascitic fluid of a Chilean GBC patient, who presented 46% European, 36% Mapuche, 12% Aymara and 6% African ancestry. Results: After immunocytochemical staining of the primary cell culture, we isolated and comprehensively characterized three independent clones (PUC-GBC1, PUC-GBC2 and PUC-GBC3) by short tandem repeat DNA profiling and RNA sequencing as well as karyotype, doubling time, chemosensitivity, in vitro migration capability and in vivo tumorigenicity assay. Primary culture cells showed high expression of CK7, CK19, CA 19-9, MUC1 and MUC16, and negative expression of mesothelial markers. The three isolated clones displayed an epithelial phenotype and an abnormal structure and number of chromosomes. RNA sequencing confirmed the increased expression of cytokeratin and mucin genes, and also of TP53 and ERBB2 with some differences among the three cells lines, and revealed a novel exonic mutation in NF1. The PUC-GBC3 clone was the most aggressive according to histopathological features and the tumorigenic capacity in NSG mice. Conclusions: The first cell lines established from a Chilean GBC patient represent a new model for studying GBC in patients of Native American descent.

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