Department of Pathology, Faculty of Medicine, Pontificia Universidad Católica de Chile, Santiago, Chile.
Applied Molecular and Cellular Biology PhD Program, Universidad de La Frontera, Temuco, Chile.
Biol Res. 2020 Apr 15;53(1):13. doi: 10.1186/s40659-020-00282-7.
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.
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.
The first cell lines established from a Chilean GBC patient represent a new model for studying GBC in patients of Native American descent.
胆囊癌（GBC）是胆道系统最常见的肿瘤。GBC 的发病率具有很大的地理变异性，在美洲原住民人群中尤为常见。在智利，GBC 是女性癌症相关死亡的第二大原因。我们在此描述了从一名智利 GBC 患者的腹水建立的三个新的细胞系，该患者具有 46%的欧洲人、36%的马普切人、12%的艾玛拉人和 6%的非洲人血统。
在原代细胞培养的免疫细胞化学染色后，我们通过短串联重复 DNA 分析和 RNA 测序以及核型、倍增时间、化疗敏感性、体外迁移能力和体内致瘤性测定，分离并全面表征了三个独立的克隆（PUC-GBC1、PUC-GBC2 和 PUC-GBC3）。原代培养细胞高表达 CK7、CK19、CA 19-9、MUC1 和 MUC16，且阴性表达间皮标志物。三个分离的克隆显示出上皮表型和异常的染色体结构和数量。RNA 测序证实了细胞角蛋白和粘蛋白基因的表达增加，以及 TP53 和 ERBB2 的表达增加，三个细胞系之间存在一些差异，并揭示了 NF1 中的一个新的外显子突变。根据组织病理学特征和 NSG 小鼠的致瘤能力，PUC-GBC3 克隆是最具侵袭性的。
从智利 GBC 患者建立的第一批细胞系为研究美洲原住民血统的 GBC 患者提供了一个新的模型。
