SUMOylation Protects FASN Against Proteasomal Degradation in Breast Cancer Cells Treated with Grape Leaf Extract

Existing therapeutic strategies for breast cancer are limited by tumor recurrence and drug-resistance. Antioxidant plant-derived compounds such as flavonoids reduce adverse outcomes and have been identified as a potential source of antineoplastic agent with less undesirable side effects. Here, we de...

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Main Authors: Andrea Floris, Michael Mazarei, Xi Yang, Aaron Elias Robinson, Jennifer Zhou, Antonio Barberis, Guy D’hallewin, Emanuela Azara, Ylenia Spissu, Ainhoa Iglesias-Ara, Sandro Orrù, Maria Lauda Tomasi
Format: Article
Language:English
Published: MDPI AG 2020-03-01
Series:Biomolecules
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Online Access:https://www.mdpi.com/2218-273X/10/4/529
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author Andrea Floris
Michael Mazarei
Xi Yang
Aaron Elias Robinson
Jennifer Zhou
Antonio Barberis
Guy D’hallewin
Emanuela Azara
Ylenia Spissu
Ainhoa Iglesias-Ara
Sandro Orrù
Maria Lauda Tomasi
author_facet Andrea Floris
Michael Mazarei
Xi Yang
Aaron Elias Robinson
Jennifer Zhou
Antonio Barberis
Guy D’hallewin
Emanuela Azara
Ylenia Spissu
Ainhoa Iglesias-Ara
Sandro Orrù
Maria Lauda Tomasi
author_sort Andrea Floris
collection DOAJ
description Existing therapeutic strategies for breast cancer are limited by tumor recurrence and drug-resistance. Antioxidant plant-derived compounds such as flavonoids reduce adverse outcomes and have been identified as a potential source of antineoplastic agent with less undesirable side effects. Here, we describe the novel regulation of fatty-acid synthase (FASN), the key enzyme in de novo fatty-acid synthesis, whereby <i>Vitis vinifera</i> L. cv Vermentino leaf hydroalcoholic extract lowers its protein stability that is regulated by small ubiquitin-like modifier (SUMO)ylation. The phenolic compounds characterization was performed by liquid chromatography–mass spectrometry (LC–MS), whereas mass spectrometry (LC–MS/MS), Western blotting/co-immunoprecipitation (Co-IP) and RT-PCR, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), clonogenicity assays, and FACS analysis were used to measure the expression of targets and tumorigenicity. Vermentino extract exhibits antitumorigenic effects, and we went on to determine that FASN and ubiquitin-conjugating enzyme 9 (UBC9), the sole E2 enzyme required for SUMOylation, were significantly reduced. Moreover, FASN was found SUMOylated in human breast cancer tissues and cell lines, and lack of SUMOylation caused by SUMO2 silencing reduced FASN protein stability. These results suggest that SUMOylation protects FASN against proteasomal degradation and may exert oncogenic activity through alteration of lipid metabolism, whereas Vermentino extract inhibits these effects which supports the additional validation of the therapeutic value of this compound in breast cancer.
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spelling doaj.art-736950741b6443fd84243eaf2c7876212023-11-19T20:14:03ZengMDPI AGBiomolecules2218-273X2020-03-0110452910.3390/biom10040529SUMOylation Protects FASN Against Proteasomal Degradation in Breast Cancer Cells Treated with Grape Leaf ExtractAndrea Floris0Michael Mazarei1Xi Yang2Aaron Elias Robinson3Jennifer Zhou4Antonio Barberis5Guy D’hallewin6Emanuela Azara7Ylenia Spissu8Ainhoa Iglesias-Ara9Sandro Orrù10Maria Lauda Tomasi11Department of Medicine, Los Angeles, CA 90048, USADepartment of Medicine, Los Angeles, CA 90048, USADepartment of Medicine, Los Angeles, CA 90048, USACedars-Sinai Medical Center, Los Angeles, CA 90048, USADepartment of Medicine, Los Angeles, CA 90048, USAInstitute of Sciences of Food Production (ISPA), 07100 Sassari, ItalyInstitute of Sciences of Food Production (ISPA), 07100 Sassari, ItalyNational Research Council (CNR), 07100 Sassari, ItalyInstitute of Sciences of Food Production (ISPA), 07100 Sassari, ItalyDepartment of Genetics, Physical Anthropology and Animal Physiology, University of the Basque Country (UPV/EHU), 48940 Bilbao, SpainDepartment of Medical Sciences and Public Health, University of Cagliari, 09042 Cagliari, ItalyDepartment of Medicine, Los Angeles, CA 90048, USAExisting therapeutic strategies for breast cancer are limited by tumor recurrence and drug-resistance. Antioxidant plant-derived compounds such as flavonoids reduce adverse outcomes and have been identified as a potential source of antineoplastic agent with less undesirable side effects. Here, we describe the novel regulation of fatty-acid synthase (FASN), the key enzyme in de novo fatty-acid synthesis, whereby <i>Vitis vinifera</i> L. cv Vermentino leaf hydroalcoholic extract lowers its protein stability that is regulated by small ubiquitin-like modifier (SUMO)ylation. The phenolic compounds characterization was performed by liquid chromatography–mass spectrometry (LC–MS), whereas mass spectrometry (LC–MS/MS), Western blotting/co-immunoprecipitation (Co-IP) and RT-PCR, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), clonogenicity assays, and FACS analysis were used to measure the expression of targets and tumorigenicity. Vermentino extract exhibits antitumorigenic effects, and we went on to determine that FASN and ubiquitin-conjugating enzyme 9 (UBC9), the sole E2 enzyme required for SUMOylation, were significantly reduced. Moreover, FASN was found SUMOylated in human breast cancer tissues and cell lines, and lack of SUMOylation caused by SUMO2 silencing reduced FASN protein stability. These results suggest that SUMOylation protects FASN against proteasomal degradation and may exert oncogenic activity through alteration of lipid metabolism, whereas Vermentino extract inhibits these effects which supports the additional validation of the therapeutic value of this compound in breast cancer.https://www.mdpi.com/2218-273X/10/4/529antioxidantbreast cancerFASNlipid metabolismpolyphenolsprotein degradation
spellingShingle Andrea Floris
Michael Mazarei
Xi Yang
Aaron Elias Robinson
Jennifer Zhou
Antonio Barberis
Guy D’hallewin
Emanuela Azara
Ylenia Spissu
Ainhoa Iglesias-Ara
Sandro Orrù
Maria Lauda Tomasi
SUMOylation Protects FASN Against Proteasomal Degradation in Breast Cancer Cells Treated with Grape Leaf Extract
Biomolecules
antioxidant
breast cancer
FASN
lipid metabolism
polyphenols
protein degradation
title SUMOylation Protects FASN Against Proteasomal Degradation in Breast Cancer Cells Treated with Grape Leaf Extract
title_full SUMOylation Protects FASN Against Proteasomal Degradation in Breast Cancer Cells Treated with Grape Leaf Extract
title_fullStr SUMOylation Protects FASN Against Proteasomal Degradation in Breast Cancer Cells Treated with Grape Leaf Extract
title_full_unstemmed SUMOylation Protects FASN Against Proteasomal Degradation in Breast Cancer Cells Treated with Grape Leaf Extract
title_short SUMOylation Protects FASN Against Proteasomal Degradation in Breast Cancer Cells Treated with Grape Leaf Extract
title_sort sumoylation protects fasn against proteasomal degradation in breast cancer cells treated with grape leaf extract
topic antioxidant
breast cancer
FASN
lipid metabolism
polyphenols
protein degradation
url https://www.mdpi.com/2218-273X/10/4/529
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