Cargando…

Polymethoxylated Flavones Target Cancer Stemness and Improve the Antiproliferative Effect of 5-Fluorouracil in a 3D Cell Model of Colorectal Cancer

Polymethoxylated flavones (PMFs) from citrus fruits are reported to present anticancer potential. However, there is a lack of information regarding their effect on cancer stem cell (CSC) populations, which has been recognized as responsible for tumor initiation, relapse, and chemoresistance. In this...

Descripción completa

Detalles Bibliográficos
Autores principales: Pereira, Carolina V., Duarte, Marlene, Silva, Patrícia, Bento da Silva, Andreia, Duarte, Catarina M. M., Cifuentes, Alejandro, García-Cañas, Virginia, Bronze, Maria R., Albuquerque, Cristina, Serra, Ana Teresa
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6412836/
https://www.ncbi.nlm.nih.gov/pubmed/30717428
http://dx.doi.org/10.3390/nu11020326
_version_ 1783402698000826368
author Pereira, Carolina V.
Duarte, Marlene
Silva, Patrícia
Bento da Silva, Andreia
Duarte, Catarina M. M.
Cifuentes, Alejandro
García-Cañas, Virginia
Bronze, Maria R.
Albuquerque, Cristina
Serra, Ana Teresa
author_facet Pereira, Carolina V.
Duarte, Marlene
Silva, Patrícia
Bento da Silva, Andreia
Duarte, Catarina M. M.
Cifuentes, Alejandro
García-Cañas, Virginia
Bronze, Maria R.
Albuquerque, Cristina
Serra, Ana Teresa
author_sort Pereira, Carolina V.
collection PubMed
description Polymethoxylated flavones (PMFs) from citrus fruits are reported to present anticancer potential. However, there is a lack of information regarding their effect on cancer stem cell (CSC) populations, which has been recognized as responsible for tumor initiation, relapse, and chemoresistance. In this study, we evaluated the effect of an orange peel extract (OPE) and its main PMFs, namely, nobiletin, sinensetin, tangeretin, and scutellarein tetramethylether in targeting cell proliferation and stemness using a 3D cell model of colorectal cancer composed of HT29 cell spheroids cultured for 7 days in stirred conditions. Soft agar assay, ALDH1 activity, and relative quantitative gene expression analysis of specific biomarkers were carried out to characterize the stemness, self-renewal, and mesenchymal features of HT29 cell spheroids. Then, the impact of OPE and PMFs in reducing cell proliferation and modulating cancer stemness and self-renewal was assessed. Results showed that, when compared with monolayer cultures, HT29 cell spheroids presented higher ALDH1 activity (81.97% ± 5.27% compared to 63.55% ± 17.49% for 2D), upregulation of CD44, PROM1, SOX9, and SNAI1 genes (1.83 ± 0.34, 2.54 ± 0.51, 2.03 ± 0.15, and 6.12 ± 1.59 times) and high self-renewal capability (352 ± 55 colonies compared to 253 ± 42 for 2D). Incubation with OPE (1 mg/mL) significantly inhibited cell proliferation and modulated cancer stemness and self-renewal ability: colony formation, ALDH1 activity, and the expression of cancer stemness biomarkers PROM1 and LGR5 were significantly reduced (0.66 ± 0.15 and 0.51 ± 0.14 times, respectively). Among all PMFs, tangeretin was the most efficient in targeting the CSC population by decreasing colony formation and the expression of PROM1 and LGR5. Scutellarein tetramethylether was shown to modulate markers of mesenchymal/metastatic transition (increasing CDH1 and reducing ZEB1 and SNAI1) and nobiletin was capable of downregulating PROM1 and SNAI1 expression. Importantly, all PMFs and OPE were shown to synergistically interact with 5-fluorouracil, improving the antiproliferative response of this drug.
format Online
Article
Text
id pubmed-6412836
institution National Center for Biotechnology Information
language English
publishDate 2019
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-64128362019-04-09 Polymethoxylated Flavones Target Cancer Stemness and Improve the Antiproliferative Effect of 5-Fluorouracil in a 3D Cell Model of Colorectal Cancer Pereira, Carolina V. Duarte, Marlene Silva, Patrícia Bento da Silva, Andreia Duarte, Catarina M. M. Cifuentes, Alejandro García-Cañas, Virginia Bronze, Maria R. Albuquerque, Cristina Serra, Ana Teresa Nutrients Article Polymethoxylated flavones (PMFs) from citrus fruits are reported to present anticancer potential. However, there is a lack of information regarding their effect on cancer stem cell (CSC) populations, which has been recognized as responsible for tumor initiation, relapse, and chemoresistance. In this study, we evaluated the effect of an orange peel extract (OPE) and its main PMFs, namely, nobiletin, sinensetin, tangeretin, and scutellarein tetramethylether in targeting cell proliferation and stemness using a 3D cell model of colorectal cancer composed of HT29 cell spheroids cultured for 7 days in stirred conditions. Soft agar assay, ALDH1 activity, and relative quantitative gene expression analysis of specific biomarkers were carried out to characterize the stemness, self-renewal, and mesenchymal features of HT29 cell spheroids. Then, the impact of OPE and PMFs in reducing cell proliferation and modulating cancer stemness and self-renewal was assessed. Results showed that, when compared with monolayer cultures, HT29 cell spheroids presented higher ALDH1 activity (81.97% ± 5.27% compared to 63.55% ± 17.49% for 2D), upregulation of CD44, PROM1, SOX9, and SNAI1 genes (1.83 ± 0.34, 2.54 ± 0.51, 2.03 ± 0.15, and 6.12 ± 1.59 times) and high self-renewal capability (352 ± 55 colonies compared to 253 ± 42 for 2D). Incubation with OPE (1 mg/mL) significantly inhibited cell proliferation and modulated cancer stemness and self-renewal ability: colony formation, ALDH1 activity, and the expression of cancer stemness biomarkers PROM1 and LGR5 were significantly reduced (0.66 ± 0.15 and 0.51 ± 0.14 times, respectively). Among all PMFs, tangeretin was the most efficient in targeting the CSC population by decreasing colony formation and the expression of PROM1 and LGR5. Scutellarein tetramethylether was shown to modulate markers of mesenchymal/metastatic transition (increasing CDH1 and reducing ZEB1 and SNAI1) and nobiletin was capable of downregulating PROM1 and SNAI1 expression. Importantly, all PMFs and OPE were shown to synergistically interact with 5-fluorouracil, improving the antiproliferative response of this drug. MDPI 2019-02-02 /pmc/articles/PMC6412836/ /pubmed/30717428 http://dx.doi.org/10.3390/nu11020326 Text en © 2019 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
Pereira, Carolina V.
Duarte, Marlene
Silva, Patrícia
Bento da Silva, Andreia
Duarte, Catarina M. M.
Cifuentes, Alejandro
García-Cañas, Virginia
Bronze, Maria R.
Albuquerque, Cristina
Serra, Ana Teresa
Polymethoxylated Flavones Target Cancer Stemness and Improve the Antiproliferative Effect of 5-Fluorouracil in a 3D Cell Model of Colorectal Cancer
title Polymethoxylated Flavones Target Cancer Stemness and Improve the Antiproliferative Effect of 5-Fluorouracil in a 3D Cell Model of Colorectal Cancer
title_full Polymethoxylated Flavones Target Cancer Stemness and Improve the Antiproliferative Effect of 5-Fluorouracil in a 3D Cell Model of Colorectal Cancer
title_fullStr Polymethoxylated Flavones Target Cancer Stemness and Improve the Antiproliferative Effect of 5-Fluorouracil in a 3D Cell Model of Colorectal Cancer
title_full_unstemmed Polymethoxylated Flavones Target Cancer Stemness and Improve the Antiproliferative Effect of 5-Fluorouracil in a 3D Cell Model of Colorectal Cancer
title_short Polymethoxylated Flavones Target Cancer Stemness and Improve the Antiproliferative Effect of 5-Fluorouracil in a 3D Cell Model of Colorectal Cancer
title_sort polymethoxylated flavones target cancer stemness and improve the antiproliferative effect of 5-fluorouracil in a 3d cell model of colorectal cancer
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6412836/
https://www.ncbi.nlm.nih.gov/pubmed/30717428
http://dx.doi.org/10.3390/nu11020326
work_keys_str_mv AT pereiracarolinav polymethoxylatedflavonestargetcancerstemnessandimprovetheantiproliferativeeffectof5fluorouracilina3dcellmodelofcolorectalcancer
AT duartemarlene polymethoxylatedflavonestargetcancerstemnessandimprovetheantiproliferativeeffectof5fluorouracilina3dcellmodelofcolorectalcancer
AT silvapatricia polymethoxylatedflavonestargetcancerstemnessandimprovetheantiproliferativeeffectof5fluorouracilina3dcellmodelofcolorectalcancer
AT bentodasilvaandreia polymethoxylatedflavonestargetcancerstemnessandimprovetheantiproliferativeeffectof5fluorouracilina3dcellmodelofcolorectalcancer
AT duartecatarinamm polymethoxylatedflavonestargetcancerstemnessandimprovetheantiproliferativeeffectof5fluorouracilina3dcellmodelofcolorectalcancer
AT cifuentesalejandro polymethoxylatedflavonestargetcancerstemnessandimprovetheantiproliferativeeffectof5fluorouracilina3dcellmodelofcolorectalcancer
AT garciacanasvirginia polymethoxylatedflavonestargetcancerstemnessandimprovetheantiproliferativeeffectof5fluorouracilina3dcellmodelofcolorectalcancer
AT bronzemariar polymethoxylatedflavonestargetcancerstemnessandimprovetheantiproliferativeeffectof5fluorouracilina3dcellmodelofcolorectalcancer
AT albuquerquecristina polymethoxylatedflavonestargetcancerstemnessandimprovetheantiproliferativeeffectof5fluorouracilina3dcellmodelofcolorectalcancer
AT serraanateresa polymethoxylatedflavonestargetcancerstemnessandimprovetheantiproliferativeeffectof5fluorouracilina3dcellmodelofcolorectalcancer