Copyright
©The Author(s) 2018. Published by Baishideng Publishing Group Inc. All rights reserved.
Establishment, functional and genetic characterization of three novel patient-derived rectal cancer cell lines
Michael Gock, Christina S Mullins, Carina Bergner, Friedrich Prall, Robert Ramer, Anja Göder, Oliver H Krämer, Falko Lange, Bernd J Krause, Ernst Klar, Michael Linnebacher
Michael Gock, Ernst Klar, Department of General Surgery, University Medical Center, Rostock 18055, Germany
Christina S Mullins, Michael Linnebacher, Section of Molecular Oncology and Immunotherapy, University Medical Center, Rostock 18055, Germany
Carina Bergner, Bernd J Krause, Department of Nuclear Medicine, University Medical Center, Rostock 18055, Germany
Friedrich Prall, Institute of Pathology, University Medical Center, Rostock 18055, Germany
Robert Ramer, Institute of Pharmacology, University Medical Center, Rostock 18055, Germany
Anja Göder, Oliver H Krämer, Institute of Toxicology, University Medical Center Mainz, Mainz 55131, Germany
Falko Lange, Oscar-Langendorff-Institute of Physiology, University Medical Center, Rostock 18055, Germany
Author contributions: Gock M, Lange F, Krause BJ and Linnebacher M designed research; Mullins CS, Prall F, Ramer R, Göder A and Lange F performed research; Gock M, Prall F, Krämer OH, Lange F, Krause BJ and Linnebacher M analyzed and reviewed data; Gock M, Bergner C and Klar E provided materials necessary to perform the experiments; Gock M and Linnebacher M wrote the manuscript; Mullins CS, Krämer OH, Lange F and Krause BJ contributed parts of the manuscript and critically reviewed the final version of the manuscript.
Supported by the German Research Foundation to Oliver H Krämer, No. KR 2291/7-1.
Institutional animal care and use committee 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 to share.
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 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/
Institutional review board statement: This study was reviewed and approved by Ethikkommission an der Medizinischen Fakultät der Universität Rostock, Germany.
Correspondence to: Michael Linnebacher, PhD, Postdoc, Research Scientist, Department of General Surgery, Section of Molecular Oncology and Immunotherapy, University Medical Center, Schillingallee 35, Rostock 18055, Germany. michael.linnebacher@med.uni-rostock.de
Telephone: +49-381-4946043 Fax: +49-381-4946002
Received: August 3, 2018
Peer-review started: August 3, 2018
First decision: October 8, 2018
Revised: October 22, 2018
Accepted: November 2, 2018
Article in press: November 2, 2018
Published online: November 21, 2018
AIM
To establish patient-individual tumor models of rectal cancer for analyses of novel biomarkers, individual response prediction and individual therapy regimens.
METHODS
Establishment of cell lines was conducted by direct in vitro culturing and in vivo xenografting with subsequent in vitro culturing. Cell lines were in-depth characterized concerning morphological features, invasive and migratory behavior, phenotype, molecular profile including mutational analysis, protein expression, and confirmation of origin by DNA fingerprint. Assessment of chemosensitivity towards an extensive range of current chemotherapeutic drugs and of radiosensitivity was performed including analysis of a combined radio- and chemotherapeutic treatment. In addition, glucose metabolism was assessed with 18F-fluorodeoxyglucose (FDG) and proliferation with 18F-fluorothymidine.
RESULTS
We describe the establishment of ultra-low passage rectal cancer cell lines of three patients suffering from rectal cancer. Two cell lines (HROC126, HROC284Met) were established directly from tumor specimens while HROC239 T0 M1 was established subsequent to xenografting of the tumor. Molecular analysis classified all three cell lines as CIMP-0/ non-MSI-H (sporadic standard) type. Mutational analysis revealed following mutational profiles: HROC126: APCwt, TP53wt, KRASwt, BRAFwt, PTENwt; HROC239 T0 M1: APCmut, P53wt, KRASmut, BRAFwt, PTENmut and HROC284Met: APCwt, P53mut, KRASmut, BRAFwt, PTENmut. All cell lines could be characterized as epithelial (EpCAM+) tumor cells with equivalent morphologic features and comparable growth kinetics. The cell lines displayed a heterogeneous response toward chemotherapy, radiotherapy and their combined application. HROC126 showed a highly radio-resistant phenotype and HROC284Met was more susceptible to a combined radiochemotherapy than HROC126 and HROC239 T0 M1. Analysis of 18F-FDG uptake displayed a markedly reduced FDG uptake of all three cell lines after combined radiochemotherapy.
CONCLUSION
These newly established and in-depth characterized ultra-low passage rectal cancer cell lines provide a useful instrument for analysis of biological characteristics of rectal cancer.
Core tip: Ultra-low passage and in-depth characterized tumor models are highly desirable for basic research and assessment of individual response prediction to current or novel therapy regimens. Here, for the first time, we describe three patient-derived rectal cancer cell lines established either directly from patient’s tumor samples or after xenografting. These tumor models were characterized according to phenotype, molecular-, as well as growth and morphological features and sensitivity to chemotherapeutic drugs and radiation, including radiochemotherapy. In addition, glucose metabolism was assessed with 18F-fluorodeoxyglucose and proliferation with 18F-fluorothymidine. These cell lines provide excellent tools for basic and translational research of rectal cancers’ biological characteristics.
