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©The Author(s) 2023. Published by Baishideng Publishing Group Inc. All rights reserved.
First report on establishment and characterization of the extrahepatic cholangiocarcinoma sarcoma cell line CBC2T-2
Ning-Zu Jiang, Ming-Zhen Bai, Chong-Fei Huang, Ze-Long Ma, Ru-Yang Zhong, Wen-Kang Fu, Long Gao, Liang Tian, Ning-Ning Mi, Hai-Dong Ma, Ya-Wen Lu, Zi-Ang Zhang, Jin-Yu Zhao, Hai-Ying Yu, Bao-Ping Zhang, Xian-Zhuo Zhang, Yan-Xian Ren, Chao Zhang, Yong Zhang, Ping Yue, Yan-Yan Lin, Wen-Bo Meng
Ning-Zu Jiang, Ming-Zhen Bai, Chong-Fei Huang, Ze-Long Ma, Ru-Yang Zhong, Wen-Kang Fu, Long Gao, Liang Tian, Ning-Ning Mi, Hai-Dong Ma, Ya-Wen Lu, Zi-Ang Zhang, Jin-Yu Zhao, Hai-Ying Yu, Bao-Ping Zhang, Xian-Zhuo Zhang, Yan-Xian Ren, Chao Zhang, Yong Zhang, Ping Yue, Yan-Yan Lin, The First Clinical Medical College, Lanzhou University, Lanzhou 730000, Gansu Province, China
Ning-Zu Jiang, Ming-Zhen Bai, Chong-Fei Huang, Ze-Long Ma, Ru-Yang Zhong, Wen-Kang Fu, Long Gao, Liang Tian, Ning-Ning Mi, Hai-Dong Ma, Ya-Wen Lu, Zi-Ang Zhang, Jin-Yu Zhao, Hai-Ying Yu, Bao-Ping Zhang, Xian-Zhuo Zhang, Yan-Xian Ren, Chao Zhang, Yong Zhang, Ping Yue, Yan-Yan Lin, Department of General Surgery, The First Hospital of Lanzhou University, Lanzhou 730000, Gansu Province, China
Ning-Zu Jiang, Ming-Zhen Bai, Chong-Fei Huang, Ze-Long Ma, Ru-Yang Zhong, Wen-Kang Fu, Long Gao, Liang Tian, Ning-Ning Mi, Hai-Dong Ma, Ya-Wen Lu, Zi-Ang Zhang, Jin-Yu Zhao, Hai-Ying Yu, Bao-Ping Zhang, Xian-Zhuo Zhang, Yan-Xian Ren, Chao Zhang, Yong Zhang, Ping Yue, Yan-Yan Lin, Laboratory of Biological Therapy and Regenerative Medicine Transformation Gansu Province, The First Hospital of Lanzhou University, Lanzhou 730000, Gansu Province, China
Wen-Bo Meng, Department of General Surgery, The First Hospital of Lanzhou University and Laboratory of Biological Therapy and Regenerative Medicine Transformation Gansu Province, Lanzhou 730000, Gansu Province, China
Author contributions: Jiang NZ and Ren YX interpreted the study design; Bai MZ and Gao L supervised our study; Yue P, Lin YY, and Meng WB obtained the research fund; Huang CF, Ma ZL, and Mi NN screened the publications, performed statistics, and drafted the manuscript; Ma HD and Lu YW helped perform statistics; Fu WK, Tian L, Zhao JY, Zhang BP, Zhang XZ, Zhang C, and Zhang Y revised the manuscript; and all authors contributed to the article and approved the submitted version.
Supported by the National Natural Science Foundation of China, No. 82060551; and Lanzhou Chengguan District Science and Technology Planning Project, No. 2019JSCX0092.
Institutional review board statement: This study was approved by the Ethics Committee of the First Hospital of Lanzhou University (LDYYLL-2022-489) and informed consent was obtained from all the patients. This study adhered to the 1964 Declaration of Helsinki and its subsequent amendments and similar ethical standards.
Institutional animal care and use committee statement: All procedures involving animals were reviewed and approved by the Institutional Animal Care and Use Committee of the First Hospital of Lanzhou University.
Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article.
Data sharing statement: The datasets used and/or analysed during the current study are available from the corresponding author upon reasonable request.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript was prepared and revised according to the ARRIVE guidelines.
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: Wen-Bo Meng, MD, PhD, Doctor, Professor, Department of General Surgery, The First Hospital of Lanzhou University and Laboratory of Biological Therapy and Regenerative Medicine Transformation Gansu Province, No. 1 Donggang West Road, Chengguan District, Lanzhou 730000, Gansu Province, China.
mengwb@lzu.edu.cn
Received: August 3, 2023
Peer-review started: August 3, 2023
First decision: September 18, 2023
Revised: September 20, 2023
Accepted: October 11, 2023
Article in press: October 11, 2023
Published online: November 7, 2023
Processing time: 93 Days and 17.3 Hours
BACKGROUND
Extrahepatic cholangiocarcinoma sarcoma is extremely rare in clinical practice. These cells consist of both epithelial and mesenchymal cells. Patient-derived cell lines that maintain tumor characteristics are valuable tools for studying the molecular mechanisms associated with carcinosarcoma. However, cholangiocarcinoma sarcoma cell lines are not available in cell banks.
AIM
To establish and characterize a new extrahepatic cholangiocarcinoma sarcoma cell line, namely CBC2T-2.
METHODS
We conducted a short tandem repeat (STR) test to confirm the identity of the CBC2T-2 cell line. Furthermore, we assessed the migratory and invasive properties of the cells and performed clonogenicity assay to evaluate the ability of individual cells to form colonies. The tumorigenic potential of CBC2T-2 cells was tested in vivo using non-obese diabetic/severe combined immunodeficient (NOD/SCID) mice. The cells were injected subcutaneously and tumor formation was observed. In addition, immunohistochemical analysis was carried out to examine the expression of epithelial marker CK19 and mesenchymal marker vimentin in both CBC2T-2 cells and xenografts. The CBC2T-2 cell line was used to screen the potential therapeutic effects of various clinical agents in patients with cholangiocarcinoma sarcoma. Lastly, whole-exome sequencing was performed to identify genetic alterations and screen for somatic mutations in the CBC2T-2 cell line.
RESULTS
The STR test showed that there was no cross-contamination and the results were identical to those of the original tissue. The cells showed round or oval-shaped epithelioid cells and mesenchymal cells with spindle-shaped or elongated morphology. The cells exhibited a high proliferation ratio with a doubling time of 47.11 h. This cell line has migratory, invasive, and clonogenic abilities. The chromosomes in the CBC2T-2 cells were polyploidy, with numbers ranging from 69 to 79. The subcutaneous tumorigenic assay confirmed the in vivo tumorigenic ability of CBC2T-2 cells in NOD/SCID mice. CBC2T-2 cells and xenografts were positive for both the epithelial marker, CK19, and the mesenchymal marker, vimentin. These results suggest that CBC2T-2 cells may have both epithelial and mesenchymal characteristics. The cells were also used to screen clinical agents in patients with cholangiocarcinoma sarcoma, and a combination of paclitaxel and gemcitabine was found to be the most effective treatment option.
CONCLUSION
We established the first human cholangiocarcinoma sarcoma cell line, CBC2T-2, with stable biogenetic traits. This cell line, as a research model, has a high clinical value and would facilitate the understanding of the pathogenesis of cholangiocarcinoma sarcoma.
Core Tip: The study established and characterized the CBC2T-2 cell line as a potential model for studying human extrahepatic cholangiocarcinoma sarcomas. The cells exhibited both epithelial and mesenchymal characteristics and demonstrated high proliferation, migration, invasion, and clonogenic abilities. Chromosomal analysis revealed polyploidy with varying chromosome numbers and in vivo tumorigenicity was confirmed in non-obese diabetic/severe combined immunodeficient mice. Immunohistochemistry indicated positive expression for both epithelial marker CK19 and mesenchymal marker vimentin. The cell line was also used to screen clinical agents, and paclitaxel and gemcitabine exhibited optimal effects. Whole-exome sequencing further revealed genetic insights.
