Establishment and characterization of patient-derived primary cell lines as preclinical models for gallbladder carcinoma

Feiling Feng; Chuncui Huang; Mingjia Xiao; Huizhen Wang; Qingxiang Gao; Zishuo Chen; Xiaoya Xu; Jun Zhou; Fugen Li; Yan Li; Dadong Zhang; Yanxin Chang; Xiaoqing Jiang

doi:10.21037/tcr.2020.02.04

Establishment and characterization of patient-derived primary cell lines as preclinical models for gallbladder carcinoma

Transl Cancer Res. 2020 Mar;9(3):1698-1710. doi: 10.21037/tcr.2020.02.04.

Authors

Feiling Feng^#¹, Chuncui Huang^#², Mingjia Xiao^#¹, Huizhen Wang^#³, Qingxiang Gao¹, Zishuo Chen³, Xiaoya Xu³, Jun Zhou³, Fugen Li³, Yan Li⁴, Dadong Zhang³, Yanxin Chang⁵, Xiaoqing Jiang¹

Affiliations

¹ Department of Biliary I, Shanghai Eastern Hepatobiliary Surgery Hospital, Navy Military Medical University, Shanghai 200438, China.
² Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.
³ Research and Development Institute of Precision Medicine, 3D Medicines Inc., Shanghai 201114, China.
⁴ University of Chinese Academy of Sciences, Beijing 100049, China.
⁵ Biliary Tract Surgery Department, Shanghai Eastern Hepatobiliary Surgery Hospital, Navy Military Medical University, Shanghai 200438, China.

^# Contributed equally.

Abstract

Background: Gallbladder carcinoma (GBC) is one of the most lethal malignancies which do not have a targeted drug in the clinic. Patient-derived primary cell lines (PDCs) are useful in assessment of cancer complexity and heterogeneity, drug-sensitivity tests, and personalized-drug-selection guidance. The aim of this study is to establish GBC PDCs and characterize their biological features.

Methods: The characterization of PDCs was defined by morphology, growth kinetics, chromosomal analysis, short tandem repeat (STR) analysis, RNA-seq and tumorigenicity. Glycosylation of PDCs derived from GBC was first studied, and the PDC model's performance were also tested and evaluated using seven molecular target inhibitors.

Results: Three novel GBC cell lines from three GBC patients were successfully established and denoted as JXQ-3D-902R4, JXQ-3D-4494R, and JXQ-3D-4786R. These cell lines demonstrated the heterogeneous characteristics of tumor morphology and phenotypes which are consistent with primary GBC, such as irregular cell shape, varied chromosomal numbers, and different STR patterns. Moreover, the growth activity and tumorigenicity ability varied among the cell lines, of which JXQ-3D-4494R exhibited the best growth rate. Furthermore, glycan profiling of whole proteins were detected and characterized. Unique N-glycans of each PDC were identified, JXQ-3D-902R4, JXQ-3D-4494R and JXQ-3D-4786R contained ten, four and seven unique glycans, respectively. The epithelial origins of three PDCs were confirmed using RNA-seq based on the highly expressed typical epithelial marker genes. Moreover, the drug-sensitivity results demonstrated that the three PDCs exhibited different responses to the seven-most commonly used targeted medicines belonging to three groups: cell-cycle inhibitors, PI3K/AKT/mTOR signaling-pathway inhibitors, and ErbB inhibitors. JXQ-3D-4494R was sensitive to most of the inhibitors, JXQ-3D-4786R was sensitive to ErbB inhibitors, and JXQ-3D-902R4 was sensitive to PI3K/AKT/mTOR inhibitors.

Conclusions: These results indicate that PDCs may be efficient preclinical models for further investigation of the biological behaviors and potential targeted therapies of human GBC.

Keywords: Gallbladder carcinoma (GBC); drug sensitivity; glycosylation; patient-derived primary cell line (PDC); preclinical model.