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©The Author(s) 2025. Published by Baishideng Publishing Group Inc. All rights reserved.
Establishment and characterization of a new human gallbladder cancer cell line, OCUG-2
Qiang Wang, Canfeng Fan, Gen Tsujio, Takashi Sakuma, Koji Maruo, Yurie Yamamoto, Daiki Imanishi, Kyoka Kawabata, Hinano Nishikubo, Saki Kanei, Rika Aoyama, Syuhei Kushiyama, Masaichi Ohira, Masakazu Yashiro
Qiang Wang, Canfeng Fan, Gen Tsujio, Takashi Sakuma, Koji Maruo, Yurie Yamamoto, Daiki Imanishi, Kyoka Kawabata, Hinano Nishikubo, Saki Kanei, Rika Aoyama, Syuhei Kushiyama, Masaichi Ohira, Masakazu Yashiro, Department of Molecular Oncology and Therapeutics, Osaka Metropolitan University Graduate School of Medicine, Osaka 545-0051, Japan
Qiang Wang, Canfeng Fan, Gen Tsujio, Takashi Sakuma, Koji Maruo, Yurie Yamamoto, Kyoka Kawabata, Hinano Nishikubo, Saki Kanei, Rika Aoyama, Syuhei Kushiyama, Masaichi Ohira, Masakazu Yashiro, Cancer Center for Translational Research, Osaka Metropolitan University Graduate School of Medicine, Osaka 545-0051, Japan
Gen Tsujio, Takashi Sakuma, Koji Maruo, Daiki Imanishi, Syuhei Kushiyama, Masaichi Ohira, Masakazu Yashiro, Department of Gastroenterological Surgery, Osaka Metropolitan University Graduate School of Medicine, Osaka 545-0051, Japan
Author contributions: Wang Q and Yashiro M designed and performed the experiments; Fan C, Tsujio G, Sakuma T, Maruo K, and Yamamoto Y performed the sample preparation; Imanishi D, Kawabata K, Nishikubo H, Kanei S, Aoyama R, and Kushiyama S conducted the material sampling; Ohira M performed the data collection; Yashiro M performed the manuscript review. All authors read and approved the final manuscript.
Institutional review board statement: The patient provided written informed consent in accordance with our IRB-approved protocol by Osaka Metropolitan University Graduate School of Medicine (approval number # 0924 and # 4391).
Institutional animal care and use committee statement: The principles of laboratory animal care and animal use were observed for the present study with the approval of the Ethical Committee for Animal Research of Osaka Metropolitan University Graduate School of Medicine (Protocol # 19001).
Conflict-of-interest statement: The authors declare that there is no conflict of interest regarding the publication of this paper.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript was prepared and revised according to the ARRIVE guidelines.
Data sharing statement: sharing statement: All relevant data are within the manuscript and its Supporting Information file.
Open Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See:
https://creativecommons.org/Licenses/by-nc/4.0/ Corresponding author: Masakazu Yashiro, MD, PhD, Associate Professor, Department of Molecular Oncology and Therapeutics, Osaka Metropolitan University Graduate School of Medicine, 1-4-3 Asahi-machi, Abeno-ku, Osaka 545-0051, Japan.
i21496f@omu.ac.jp
Received: August 20, 2024
Revised: November 19, 2024
Accepted: January 7, 2025
Published online: June 20, 2025
Processing time: 242 Days and 18.9 Hours
BACKGROUND
Gallbladder cancer (GBC) is a highly aggressive malignant tumor originating from the biliary tract. As one of the most common malignancies in the biliary system, GBC is particularly challenging due to its tendency to remain asymptomatic, which often results in delayed diagnoses even at advanced stages. Combined with its invasive potential and poor response to conventional therapies, GBC has a high mortality rate, highlighting the critical need for innovative therapeutic approaches. Identifying molecular biomarkers for early detection and discovering novel therapeutic targets might be essential to improving outcomes of patients with GBC.
AIM
To establish a novel GBC cell line to investigate the molecular mechanisms underlying GBC progression and evaluate potential therapeutic targets.
METHODS
We developed a unique GBC cell line, named OCUG-2, derived from a metastatic peritoneal implant, and verified its authenticity using short tandem repeat (STR) profiling. RT-PCR and RNA sequencing (RNA-seq) were performed to assess gene expression profiles, with functional enrichment analyzed through Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses. The MTT cell proliferation assay and an invasion assay were performed to evaluate response to nine inhibitors. Immunohistochemistry (IHC) was conducted on 34 GBC samples to analyze insulin-like growth factor 1 receptor (IGF1R) expression.
RESULTS
OCUG-2 cells displayed adhesive growth with dendritic morphology and a 30-hour doubling time. Subcutaneous inoculation of OCUG-2 cells into mice confirmed their tumorigenic potential. STR analysis authenticated the cell line, and there was high mRNA and protein expression of IGF1R in OCUG-2 cells. The IGF1R inhibitor picropodophyllotoxin significantly inhibited OCUG-2 cell proliferation, yielding an IC50 of 0.49 μM. RNA-seq analysis identified gene fusions, and GO/KEGG functional enrichment analyses revealed pathways implicated in cancer progression. IHC analysis showed IGF1R positivity in 18 of 34 GBC cases, with significant association between IGF1R expression and poor prognosis. In invasion assays, an IGF1R inhibitor effectively reduced OCUG-2 cell invasiveness.
CONCLUSION
IGF1R might be a promising target for GBC. The newly established OCUG-2 cell line serves as a valuable model for investigating the molecular mechanisms of GBC and evaluating therapeutic strategies.
Core Tip: We established a new gallbladder cancer (GBC) cell line derived from a metastatic peritoneal implant of GBC. These cells, named OCUG-2, showed adhesive growth with a dendritic morphology. Doubling time of OCUG-2 cells was 30 hours. OCUG-2 cells were positive for insulin-like growth factor 1 receptor (IGF1R), and an IGF1R inhibitor reduced the proliferation of OCUG-2 cells. Immunohistochemistry found IGF1R positivity in 18 of 34 cases of GBC, and IGF1R expression was associated with poor prognosis. These findings suggest that IGF1R may be a promising target for GBC and OCUG-2 might be useful for analysis of GBC.
