Cargando…
Establishment and Characterization of the Novel High-Grade Serous Ovarian Cancer Cell Line OVPA8
High-grade serous ovarian carcinoma (HGSOC) is the most frequent histological type of ovarian cancer and the one with worst prognosis. Unfortunately, the majority of established ovarian cancer cell lines which are used in the research have unclear histological origin and probably do not represent HG...
Autores principales: | , , , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2018
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6073376/ https://www.ncbi.nlm.nih.gov/pubmed/30018258 http://dx.doi.org/10.3390/ijms19072080 |
_version_ | 1783344175891087360 |
---|---|
author | Tudrej, Patrycja Olbryt, Magdalena Zembala-Nożyńska, Ewa Kujawa, Katarzyna A. Cortez, Alexander J. Fiszer-Kierzkowska, Anna Pigłowski, Wojciech Nikiel, Barbara Głowala-Kosińska, Magdalena Bartkowska-Chrobok, Aleksandra Smagur, Andrzej Fidyk, Wojciech Lisowska, Katarzyna M. |
author_facet | Tudrej, Patrycja Olbryt, Magdalena Zembala-Nożyńska, Ewa Kujawa, Katarzyna A. Cortez, Alexander J. Fiszer-Kierzkowska, Anna Pigłowski, Wojciech Nikiel, Barbara Głowala-Kosińska, Magdalena Bartkowska-Chrobok, Aleksandra Smagur, Andrzej Fidyk, Wojciech Lisowska, Katarzyna M. |
author_sort | Tudrej, Patrycja |
collection | PubMed |
description | High-grade serous ovarian carcinoma (HGSOC) is the most frequent histological type of ovarian cancer and the one with worst prognosis. Unfortunately, the majority of established ovarian cancer cell lines which are used in the research have unclear histological origin and probably do not represent HGSOC. Thus, new and reliable models of HGSOC are needed. Ascitic fluid from a patient with recurrent HGSOC was used to establish a stable cancer cell line. Cells were characterized by cytogenetic karyotyping and short tandem repeat (STR) profiling. New generation sequencing was applied to test for hot-spot mutations in 50 cancer-associated genes and fluorescence in situ hybridization (FISH) analysis was used to check for TP53 status. Cells were analyzed for expression of several marker genes/proteins by reverse-transcription polymerase chain reaction (RT-PCR), fluorescence-activated cell sorting (FACS), and immunocytochemistry (ICC). Functional tests were performed to compare OVPA8 cells with five commercially available and frequently used ovarian cancer cell lines: SKOV3, A2780, OVCAR3, ES2, and OAW42. Our newly-established OVPA8 cell line shows morphologic and genetic features consistent with HGSOC, such as epithelial morphology, multiple chromosomal aberrations, TP53 mutation, BRCA1 mutation, and loss of one copy of BRCA2. The OVPA8 line has a stable STR profile. Cells are positive for EpCAM, CK19, and CD44; they have relatively low plating efficiency/ability to form spheroids, a low migration rate, and intermediate invasiveness in matrigel, as compared to other ovarian cancer lines. OVPA8 is sensitive to paclitaxel and resistant to cisplatin. We also tested two FGFR inhibitors; OVPA8 cells were resistant to AZD4547 (AstraZeneca, London, UK), but sensitive to the new inhibitor CPL304-110-01 (Celon Pharma, Łomianki/Kiełpin, Poland). We have established and characterized a novel cell line, OVPA8, which can be a valuable preclinical model for studies on high-grade serous ovarian cancer. |
format | Online Article Text |
id | pubmed-6073376 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-60733762018-08-13 Establishment and Characterization of the Novel High-Grade Serous Ovarian Cancer Cell Line OVPA8 Tudrej, Patrycja Olbryt, Magdalena Zembala-Nożyńska, Ewa Kujawa, Katarzyna A. Cortez, Alexander J. Fiszer-Kierzkowska, Anna Pigłowski, Wojciech Nikiel, Barbara Głowala-Kosińska, Magdalena Bartkowska-Chrobok, Aleksandra Smagur, Andrzej Fidyk, Wojciech Lisowska, Katarzyna M. Int J Mol Sci Article High-grade serous ovarian carcinoma (HGSOC) is the most frequent histological type of ovarian cancer and the one with worst prognosis. Unfortunately, the majority of established ovarian cancer cell lines which are used in the research have unclear histological origin and probably do not represent HGSOC. Thus, new and reliable models of HGSOC are needed. Ascitic fluid from a patient with recurrent HGSOC was used to establish a stable cancer cell line. Cells were characterized by cytogenetic karyotyping and short tandem repeat (STR) profiling. New generation sequencing was applied to test for hot-spot mutations in 50 cancer-associated genes and fluorescence in situ hybridization (FISH) analysis was used to check for TP53 status. Cells were analyzed for expression of several marker genes/proteins by reverse-transcription polymerase chain reaction (RT-PCR), fluorescence-activated cell sorting (FACS), and immunocytochemistry (ICC). Functional tests were performed to compare OVPA8 cells with five commercially available and frequently used ovarian cancer cell lines: SKOV3, A2780, OVCAR3, ES2, and OAW42. Our newly-established OVPA8 cell line shows morphologic and genetic features consistent with HGSOC, such as epithelial morphology, multiple chromosomal aberrations, TP53 mutation, BRCA1 mutation, and loss of one copy of BRCA2. The OVPA8 line has a stable STR profile. Cells are positive for EpCAM, CK19, and CD44; they have relatively low plating efficiency/ability to form spheroids, a low migration rate, and intermediate invasiveness in matrigel, as compared to other ovarian cancer lines. OVPA8 is sensitive to paclitaxel and resistant to cisplatin. We also tested two FGFR inhibitors; OVPA8 cells were resistant to AZD4547 (AstraZeneca, London, UK), but sensitive to the new inhibitor CPL304-110-01 (Celon Pharma, Łomianki/Kiełpin, Poland). We have established and characterized a novel cell line, OVPA8, which can be a valuable preclinical model for studies on high-grade serous ovarian cancer. MDPI 2018-07-17 /pmc/articles/PMC6073376/ /pubmed/30018258 http://dx.doi.org/10.3390/ijms19072080 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Tudrej, Patrycja Olbryt, Magdalena Zembala-Nożyńska, Ewa Kujawa, Katarzyna A. Cortez, Alexander J. Fiszer-Kierzkowska, Anna Pigłowski, Wojciech Nikiel, Barbara Głowala-Kosińska, Magdalena Bartkowska-Chrobok, Aleksandra Smagur, Andrzej Fidyk, Wojciech Lisowska, Katarzyna M. Establishment and Characterization of the Novel High-Grade Serous Ovarian Cancer Cell Line OVPA8 |
title | Establishment and Characterization of the Novel High-Grade Serous Ovarian Cancer Cell Line OVPA8 |
title_full | Establishment and Characterization of the Novel High-Grade Serous Ovarian Cancer Cell Line OVPA8 |
title_fullStr | Establishment and Characterization of the Novel High-Grade Serous Ovarian Cancer Cell Line OVPA8 |
title_full_unstemmed | Establishment and Characterization of the Novel High-Grade Serous Ovarian Cancer Cell Line OVPA8 |
title_short | Establishment and Characterization of the Novel High-Grade Serous Ovarian Cancer Cell Line OVPA8 |
title_sort | establishment and characterization of the novel high-grade serous ovarian cancer cell line ovpa8 |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6073376/ https://www.ncbi.nlm.nih.gov/pubmed/30018258 http://dx.doi.org/10.3390/ijms19072080 |
work_keys_str_mv | AT tudrejpatrycja establishmentandcharacterizationofthenovelhighgradeserousovariancancercelllineovpa8 AT olbrytmagdalena establishmentandcharacterizationofthenovelhighgradeserousovariancancercelllineovpa8 AT zembalanozynskaewa establishmentandcharacterizationofthenovelhighgradeserousovariancancercelllineovpa8 AT kujawakatarzynaa establishmentandcharacterizationofthenovelhighgradeserousovariancancercelllineovpa8 AT cortezalexanderj establishmentandcharacterizationofthenovelhighgradeserousovariancancercelllineovpa8 AT fiszerkierzkowskaanna establishmentandcharacterizationofthenovelhighgradeserousovariancancercelllineovpa8 AT pigłowskiwojciech establishmentandcharacterizationofthenovelhighgradeserousovariancancercelllineovpa8 AT nikielbarbara establishmentandcharacterizationofthenovelhighgradeserousovariancancercelllineovpa8 AT głowalakosinskamagdalena establishmentandcharacterizationofthenovelhighgradeserousovariancancercelllineovpa8 AT bartkowskachrobokaleksandra establishmentandcharacterizationofthenovelhighgradeserousovariancancercelllineovpa8 AT smagurandrzej establishmentandcharacterizationofthenovelhighgradeserousovariancancercelllineovpa8 AT fidykwojciech establishmentandcharacterizationofthenovelhighgradeserousovariancancercelllineovpa8 AT lisowskakatarzynam establishmentandcharacterizationofthenovelhighgradeserousovariancancercelllineovpa8 |