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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...

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Autores principales: 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.
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
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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.
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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
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