Basic Study
Copyright ©The Author(s) 2018. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Sep 7, 2018; 24(33): 3749-3759
Published online Sep 7, 2018. doi: 10.3748/wjg.v24.i33.3749
Establishment, functional and genetic characterization of a colon derived large cell neuroendocrine carcinoma cell line
Michael Gock, Christina S Mullins, Christine Harnack, Friedrich Prall, Robert Ramer, Anja Göder, Oliver H Krämer, Ernst Klar, Michael Linnebacher
Michael Gock, Ernst Klar, Department of General Surgery, University of Rostock, Rostock 18055, Germany
Christina S Mullins, Christine Harnack, Michael Linnebacher, Department of General Surgery, Section of Molecular Oncology and Immunotherapy, University of Rostock, Rostock 18055, Germany
Friedrich Prall, Institute of Pathology, University of Rostock, Rostock 18055, Germany
Robert Ramer, Institute of Pharmacology, University of Rostock, Rostock 18055, Germany
Anja Göder, Oliver H Krämer, Institute of Toxicology, University Medical Center Mainz, Mainz 55131, Germany
Author contributions: Linnebacher M and Klar E designed research; Mullins CS, Harnack C, Prall F, Ramer R and Göder A performed research; Gock M, Krämer OH and Linnebacher M analyzed data; Gock M, Mullins CS, Krämer OH and Linnebacher M wrote the paper.
Institutional review board statement: This study was reviewed and approved by Ethikkommission an der Medizinischen Fakultät der Universität Rostock, St.-Georg-Str. 108, Rostock 18055, Germany.
Informed consent statement: All procedures were approved by Ethikkommission an der Medizinischen Fakultät der Universität Rostock (reference number II HV 43/2004) in accordance with generally accepted guidelines for the use of human material.
Conflict-of-interest statement: To the best of our knowledge, no conflict of interest exists.
Data sharing statement: No additional data are available.
ARRIVE guidelines statement: The ARRIVE Guidelines have been adopted.
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (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: http://creativecommons.org/licenses/by-nc/4.0/
Correspondence to: Michael Linnebacher, PhD, Postdoc, Research Scientist, Department of General Surgery, Section of Molecular Oncology and Immunotherapy, University of Rostock, Schillingallee 35, Rostock 18055, Germany. michael.linnebacher@med.uni-rostock.de
Telephone: +49-381-4946043 Fax: +49-381-4946002
Received: February 28, 2018
Peer-review started: March 1, 2018
First decision: April 18, 2018
Revised: June 14, 2018
Accepted: June 16, 2018
Article in press: June 16, 2018
Published online: September 7, 2018
Processing time: 189 Days and 15.9 Hours
Abstract
AIM

To establish cell line and patient-derived xenograft (PDX) models for neuroendocrine carcinomas (NEC) which is highly desirable for gaining insight into tumor development as well as preclinical research including biomarker testing and drug response prediction.

METHODS

Cell line establishment was conducted from direct in vitro culturing of colonic NEC tissue (HROC57). A PDX could also successfully be established from vitally frozen tumor samples. Morphological features, invasive and migratory behavior of the HROC57 cells as well as expression of neuroendocrine markers were vastly analyzed. Phenotypic analysis was done by microscopy and multicolor flow cytometry. The extensive molecular-pathological profiling included mutation analysis, assessment of chromosomal and microsatellite instability; and in addition, fingerprinting (i.e., STR analysis) was performed from the cell line in direct comparison to primary patient-derived tissues and the PDX model established. Drug responsiveness was examined for a panel of chemotherapeutics in clinical use for the treatment of solid cancers.

RESULTS

The established cell line HROC57 showed distinct morphological and molecular features of a poorly differentiated large-cell NEC with KI-67 > 50%. Molecular-pathological analysis revealed a CpG island promoter methylation positive cell line with microsatellite instability being absent. The following mutation profile was observed: KRAS (wt), BRAF (mut). A high sensitivity to etoposide, cisplatin and 5-FU could be demonstrated while it was more resistant towards rapamycin.

CONCLUSION

We successfully established and characterized a novel patient-derived NEC cell line in parallel to a PDX model as a useful tool for further analysis of the biological characteristics and for development of novel diagnostic and therapeutic options for NEC.

Keywords: Patient-derived tumor model; Large cell neuroendocrine carcinoma; Individualized medicine

Core tip: Since incidence of G3 poorly differentiated neuroendocrine carcinomas (NEC) is very low, data is substantially scarcer than on G1 or G2 neuroendocrine tumors. Herein we describe an ultra-low passage NEC cell line and corresponding patient-derived xenograft model established directly from patient derived colonic tumor samples. We characterized our model according to phenotype, molecular, morphological and growth characteristics, as well as drug response and radiation response profiles. We present a useful tool for further analysis of the biological characteristics and for development of novel diagnostic and therapeutic options for NEC. The model is available on request.