Functional and transcriptomic characterization of carboplatin-resistant A2780 ovarian cancer cell lines

Abstract Background: Ovarian cancer is a significant cancer-related cause of death in women worldwide. The most used chemotherapeutic regimen is based on carboplatin (CBDCA). However, CBDCA resistance is the main obstacle to a better prognosis. An in vitro drug-resistant cell model would help in th...

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Autores principales:	Viscarra,Tamara, Buchegger,Kurt, Jofre,Ignacio, Riquelme,Ismael, Zanella,Louise, Abanto,Michel, Parker,Alyssa C., Piccolo,Stephen R., Roa,Juan Carlos, Ili,Carmen, Brebi,Priscilla
Lenguaje:	English
Publicado:	Sociedad de Biología de Chile 2019
Materias:	A2780 cell line Carboplatin Drug resistance Wnt/β-catenin-signaling pathway Integrin signaling pathway Ovarian cancer
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602019000100211
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spelling	oai:scielo:S0716-976020190001002112019-10-10Functional and transcriptomic characterization of carboplatin-resistant A2780 ovarian cancer cell linesViscarra,TamaraBuchegger,KurtJofre,IgnacioRiquelme,IsmaelZanella,LouiseAbanto,MichelParker,Alyssa C.Piccolo,Stephen R.Roa,Juan CarlosIli,CarmenBrebi,Priscilla A2780 cell line Carboplatin Drug resistance Wnt/β-catenin-signaling pathway Integrin signaling pathway Ovarian cancer Abstract Background: Ovarian cancer is a significant cancer-related cause of death in women worldwide. The most used chemotherapeutic regimen is based on carboplatin (CBDCA). However, CBDCA resistance is the main obstacle to a better prognosis. An in vitro drug-resistant cell model would help in the understanding of molecular mechanisms underlying this drug-resistance phenomenon. The aim of this study was to characterize cellular and molecular changes of induced CBDCA-resistant ovarian cancer cell line A2780. Methods: The cell selection strategy used in this study was a dose-per-pulse method using a concentration of 100 μM for 2 h. Once 20 cycles of exposure to the drug were completed, the cell cultures showed a resistant phenotype. Then, the ovarian cancer cell line A2780 was grown with 100 μM of CBDCA (CBDCA-resistant cells) or without CBDCA (parental cells). After, a drug sensitivity assay, morphological analyses, cell death assays and a RNA-seq analysis were performed in CBDCA-resistant A2780 cells. Results: Microscopy on both parental and CBDCA-resistant A2780 cells showed similar characteristics in morphology and F-actin distribution within cells. In cell-death assays, parental A2780 cells showed a significant increase in phosphatidylserine translocation and caspase-3/7 cleavage compared to CBDCA-resistant A2780 cells (P < 0.05 and P < 0.005, respectively). Cell viability in parental A2780 cells was significantly decreased compared to CBDCA-resistant A2780 cells (P < 0.0005). The RNA-seq analysis showed 156 differentially expressed genes (DEGs) associated mainly to molecular functions. Conclusion: CBDCA-resistant A2780 ovarian cancer cells is a reliable model of CBDCA resistance that shows several DEGs involved in molecular functions such as transmembrane activity, protein binding to cell surface receptor and catalytic activity. Also, we found that the Wnt/3-catenin and integrin signaling pathway are the main metabolic pathway dysregulated in CBDCA-resistant A2780 cells.info:eu-repo/semantics/openAccessSociedad de Biología de ChileBiological Research v.52 20192019-01-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602019000100211en10.1186/s40659-019-0220-0
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	A2780 cell line Carboplatin Drug resistance Wnt/β-catenin-signaling pathway Integrin signaling pathway Ovarian cancer
spellingShingle	A2780 cell line Carboplatin Drug resistance Wnt/β-catenin-signaling pathway Integrin signaling pathway Ovarian cancer Viscarra,Tamara Buchegger,Kurt Jofre,Ignacio Riquelme,Ismael Zanella,Louise Abanto,Michel Parker,Alyssa C. Piccolo,Stephen R. Roa,Juan Carlos Ili,Carmen Brebi,Priscilla Functional and transcriptomic characterization of carboplatin-resistant A2780 ovarian cancer cell lines
description	Abstract Background: Ovarian cancer is a significant cancer-related cause of death in women worldwide. The most used chemotherapeutic regimen is based on carboplatin (CBDCA). However, CBDCA resistance is the main obstacle to a better prognosis. An in vitro drug-resistant cell model would help in the understanding of molecular mechanisms underlying this drug-resistance phenomenon. The aim of this study was to characterize cellular and molecular changes of induced CBDCA-resistant ovarian cancer cell line A2780. Methods: The cell selection strategy used in this study was a dose-per-pulse method using a concentration of 100 μM for 2 h. Once 20 cycles of exposure to the drug were completed, the cell cultures showed a resistant phenotype. Then, the ovarian cancer cell line A2780 was grown with 100 μM of CBDCA (CBDCA-resistant cells) or without CBDCA (parental cells). After, a drug sensitivity assay, morphological analyses, cell death assays and a RNA-seq analysis were performed in CBDCA-resistant A2780 cells. Results: Microscopy on both parental and CBDCA-resistant A2780 cells showed similar characteristics in morphology and F-actin distribution within cells. In cell-death assays, parental A2780 cells showed a significant increase in phosphatidylserine translocation and caspase-3/7 cleavage compared to CBDCA-resistant A2780 cells (P < 0.05 and P < 0.005, respectively). Cell viability in parental A2780 cells was significantly decreased compared to CBDCA-resistant A2780 cells (P < 0.0005). The RNA-seq analysis showed 156 differentially expressed genes (DEGs) associated mainly to molecular functions. Conclusion: CBDCA-resistant A2780 ovarian cancer cells is a reliable model of CBDCA resistance that shows several DEGs involved in molecular functions such as transmembrane activity, protein binding to cell surface receptor and catalytic activity. Also, we found that the Wnt/3-catenin and integrin signaling pathway are the main metabolic pathway dysregulated in CBDCA-resistant A2780 cells.
author	Viscarra,Tamara Buchegger,Kurt Jofre,Ignacio Riquelme,Ismael Zanella,Louise Abanto,Michel Parker,Alyssa C. Piccolo,Stephen R. Roa,Juan Carlos Ili,Carmen Brebi,Priscilla
author_facet	Viscarra,Tamara Buchegger,Kurt Jofre,Ignacio Riquelme,Ismael Zanella,Louise Abanto,Michel Parker,Alyssa C. Piccolo,Stephen R. Roa,Juan Carlos Ili,Carmen Brebi,Priscilla
author_sort	Viscarra,Tamara
title	Functional and transcriptomic characterization of carboplatin-resistant A2780 ovarian cancer cell lines
title_short	Functional and transcriptomic characterization of carboplatin-resistant A2780 ovarian cancer cell lines
title_full	Functional and transcriptomic characterization of carboplatin-resistant A2780 ovarian cancer cell lines
title_fullStr	Functional and transcriptomic characterization of carboplatin-resistant A2780 ovarian cancer cell lines
title_full_unstemmed	Functional and transcriptomic characterization of carboplatin-resistant A2780 ovarian cancer cell lines
title_sort	functional and transcriptomic characterization of carboplatin-resistant a2780 ovarian cancer cell lines
publisher	Sociedad de Biología de Chile
publishDate	2019
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602019000100211
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Functional and transcriptomic characterization of carboplatin-resistant A2780 ovarian cancer cell lines

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