Basic Study
Copyright ©The Author(s) 2023. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. May 7, 2023; 29(17): 2642-2656
Published online May 7, 2023. doi: 10.3748/wjg.v29.i17.2642
Establishment and characterization of a new human ampullary carcinoma cell line, DPC-X1
Hao Xu, Chang-Peng Chai, Xin Miao, Huan Tang, Jin-Jing Hu, Hui Zhang, Wen-Ce Zhou
Hao Xu, Chang-Peng Chai, Jin-Jing Hu, The Forth Department of General Surgery, The First Hospital of Lanzhou University, Lanzhou 730000, Gansu Province, China
Xin Miao, State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Animal Virology of the Ministry of Agriculture, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730000, Gansu Province, China
Huan Tang, Wen-Ce Zhou, The Second Clinical Medical College, Lanzhou University, Lanzhou 730000, Gansu Province, China
Hui Zhang, Wen-Ce Zhou, Department of General Surgery, The Second Hospital of Lanzhou University, Lanzhou 730000, Gansu Province, China
Author contributions: Xu H designed and coordinated the study; Xu H, Chai CP, Miao X and Tang H performed the experiments, acquired and analyzed data; Xu H, Hu JJ and Zhang H interpreted the data; Xu H, Chai CP and Zhou WC wrote the manuscript; All authors have read and approve the final manuscript.
Supported by National Natural Science Foundation of China, No. 82260555; Gansu Provincial Science and Technology Plan, No. 1606RJZA139, No. 21JR11RA096, No. 21JR1RA099, and No. 21JR1RA113; Gansu Health Industry Project, No. GSWSKY2020-21; Traditional Chinese Medicine Scientific Research Project of Gansu Province, No. GZKP-2020-28; Talent Innovation and Entrepreneurship Project of Lanzhou, No. 2020-RC-46; Intra-Hospital Fund of the First Hospital of Lanzhou University, No. ldyyyn2019-97.
Institutional review board statement: The study was conducted according to the guidelines of the Declaration of Helsinki, and approved by the Ethics Committee of the First Hospital of Lanzhou University, No. LDYYLL-2022-487.
Institutional animal care and use committee statement: The animal study was conducted according to the guidelines of the Declaration of Helsinki, and approved by the Ethics Committee of the First Hospital of Lanzhou University, No. LDYYLL-2022-487.
Informed consent statement: All study participants or their legal guardian provided informed written consent about personal and medical data collection prior to study enrolment.
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: Hao Xu, Doctor, MD, Associate Chief Physician, Surgeon, The Forth Department of General Surgery, The First Hospital of Lanzhou University, No. 1 Donggang West Road, Lanzhou 730000, Gansu Province, China. amoyxu@126.com
Received: December 26, 2022
Peer-review started: December 26, 2022
First decision: February 8, 2023
Revised: February 17, 2023
Accepted: April 13, 2023
Article in press: April 13, 2023
Published online: May 7, 2023
Processing time: 132 Days and 2.8 Hours
Abstract
BACKGROUND

An in-depth study of the pathogenesis and biological characteristics of ampullary carcinoma is necessary to identify appropriate treatment strategies. To date, only eight ampullary cancer cell lines have been reported, and a mixed-type ampullary carcinoma cell line has not yet been reported.

AIM

To establish a stable mixed-type ampullary carcinoma cell line originating from Chinese.

METHODS

Fresh ampullary cancer tissue samples were used for primary culture and subculture. The cell line was evaluated by cell proliferation assays, clonal formation assays, karyotype analysis, short tandem repeat (STR) analysis and transmission electron microscopy. Drug resistances against oxaliplatin, paclitaxel, gemcitabine and 5-FU were evaluated by cell counting kit-8 assay. Subcutaneous injection 1 × 106 cells to three BALB/c nude mice for xenograft studies. The hematoxylin-eosin staining was used to detect the pathological status of the cell line. The expression of biomarkers cytokeratin 7 (CK7), cytokeratin 20 (CK20), cytokeratin low molecular weight (CKL), Ki67 and carcinoembryonic antigen (CEA) were determined by immunocytochemistry assay.

RESULTS

DPC-X1 was continuously cultivated for over a year and stably passaged for more than 80 generations; its population doubling time was 48 h. STR analysis demonstrated that the characteristics of DPC-X1 were highly consistent with those of the patient’s primary tumor. Furthermore, karyotype analysis revealed its abnormal sub-tetraploid karyotype. DPC-X1 could efficiently form organoids in suspension culture. Under the transmission electron microscope, microvilli and pseudopods were observed on the cell surface, and desmosomes were visible between the cells. DPC-X1 cells inoculated into BALB/C nude mice quickly formed transplanted tumors, with a tumor formation rate of 100%. Their pathological characteristics were similar to those of the primary tumor. Moreover, DPC-X1 was sensitive to oxaliplatin and paclitaxel and resistant to gemcitabine and 5-FU. Immunohistochemistry showed that the DPC-X1 cells were strongly positive for CK7, CK20, and CKL; the Ki67 was 50%, and CEA was focally expressed.

CONCLUSION

Here, we have constructed a mixed-type ampullary carcinoma cell line that can be used as an effective model for studying the pathogenesis of ampullary carcinoma and drug development.

Keywords: Ampullary carcinoma; Cell line; Xenograft; Drug resistance

Core Tip: A new ampullary carcinoma cell line has been established, making up for the shortage of Chinese ampullary carcinoma cell lines. And this is the the first study to report a mixed-type ampullary carcinoma cell line. The cell line inoculated into BALB/c nude mice can quickly form xenograft tumors. It is an excellent model for studying the mechanisms of invasion, metastasis and other mechanisms of ampullary carcinoma. The cell line is sensitive to oxaliplatin and paclitaxel but is naturally resistant to gemcitabine and fluorouracil, which can be used for drug resistance mechanism research and new drug development.