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Inhibition of ABCC6 Transporter Modifies Cytoskeleton and Reduces Motility of HepG2 Cells via Purinergic Pathway

ABCC6, belonging to sub-family C of ATP-binding cassette transporter, is an ATP-dependent transporter mainly present in the basolateral plasma membrane of hepatic and kidney cells. Although the substrates transported are still uncertain, ABCC6 has been shown to promote ATP release. The extracellular...

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Autores principales: Ostuni, Angela, Carmosino, Monica, Miglionico, Rocchina, Abruzzese, Vittorio, Martinelli, Fabio, Russo, Daniela, Laurenzana, Ilaria, Petillo, Agata, Bisaccia, Faustino
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7349786/
https://www.ncbi.nlm.nih.gov/pubmed/32517079
http://dx.doi.org/10.3390/cells9061410
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author Ostuni, Angela
Carmosino, Monica
Miglionico, Rocchina
Abruzzese, Vittorio
Martinelli, Fabio
Russo, Daniela
Laurenzana, Ilaria
Petillo, Agata
Bisaccia, Faustino
author_facet Ostuni, Angela
Carmosino, Monica
Miglionico, Rocchina
Abruzzese, Vittorio
Martinelli, Fabio
Russo, Daniela
Laurenzana, Ilaria
Petillo, Agata
Bisaccia, Faustino
author_sort Ostuni, Angela
collection PubMed
description ABCC6, belonging to sub-family C of ATP-binding cassette transporter, is an ATP-dependent transporter mainly present in the basolateral plasma membrane of hepatic and kidney cells. Although the substrates transported are still uncertain, ABCC6 has been shown to promote ATP release. The extracellular ATP and its derivatives di- and mono-nucleotides and adenosine by acting on specific receptors activate the so-called purinergic pathway, which in turn controls relevant cellular functions such as cell immunity, inflammation, and cancer. Here, we analyzed the effect of Abcc6 knockdown and probenecid-induced ABCC6 inhibition on cell cycle, cytoskeleton, and motility of HepG2 cells. Gene and protein expression were evaluated by quantitative Reverse Transcription PCR (RT-qPCR) and western blot, respectively. Cellular cycle analysis was evaluated by flow cytometry. Actin cytoskeleton dynamics was evaluated by laser confocal microscopy using fluorophore-conjugated phalloidin. Cell motility was analyzed by in vitro wound-healing migration assay. Cell migration is reduced both in Abcc6 knockdown HepG2 cells and in probenecid treated HepG2 cells by interfering with the extracellular reserve of ATP. Therefore, ABCC6 could contribute to cytoskeleton rearrangements and cell motility through purinergic signaling. Altogether, our findings shed light on a new role of the ABCC6 transporter in HepG2 cells and suggest that its inhibitor/s could be considered potential anti-metastatic drugs.
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spelling pubmed-73497862020-07-15 Inhibition of ABCC6 Transporter Modifies Cytoskeleton and Reduces Motility of HepG2 Cells via Purinergic Pathway Ostuni, Angela Carmosino, Monica Miglionico, Rocchina Abruzzese, Vittorio Martinelli, Fabio Russo, Daniela Laurenzana, Ilaria Petillo, Agata Bisaccia, Faustino Cells Article ABCC6, belonging to sub-family C of ATP-binding cassette transporter, is an ATP-dependent transporter mainly present in the basolateral plasma membrane of hepatic and kidney cells. Although the substrates transported are still uncertain, ABCC6 has been shown to promote ATP release. The extracellular ATP and its derivatives di- and mono-nucleotides and adenosine by acting on specific receptors activate the so-called purinergic pathway, which in turn controls relevant cellular functions such as cell immunity, inflammation, and cancer. Here, we analyzed the effect of Abcc6 knockdown and probenecid-induced ABCC6 inhibition on cell cycle, cytoskeleton, and motility of HepG2 cells. Gene and protein expression were evaluated by quantitative Reverse Transcription PCR (RT-qPCR) and western blot, respectively. Cellular cycle analysis was evaluated by flow cytometry. Actin cytoskeleton dynamics was evaluated by laser confocal microscopy using fluorophore-conjugated phalloidin. Cell motility was analyzed by in vitro wound-healing migration assay. Cell migration is reduced both in Abcc6 knockdown HepG2 cells and in probenecid treated HepG2 cells by interfering with the extracellular reserve of ATP. Therefore, ABCC6 could contribute to cytoskeleton rearrangements and cell motility through purinergic signaling. Altogether, our findings shed light on a new role of the ABCC6 transporter in HepG2 cells and suggest that its inhibitor/s could be considered potential anti-metastatic drugs. MDPI 2020-06-05 /pmc/articles/PMC7349786/ /pubmed/32517079 http://dx.doi.org/10.3390/cells9061410 Text en © 2020 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
Ostuni, Angela
Carmosino, Monica
Miglionico, Rocchina
Abruzzese, Vittorio
Martinelli, Fabio
Russo, Daniela
Laurenzana, Ilaria
Petillo, Agata
Bisaccia, Faustino
Inhibition of ABCC6 Transporter Modifies Cytoskeleton and Reduces Motility of HepG2 Cells via Purinergic Pathway
title Inhibition of ABCC6 Transporter Modifies Cytoskeleton and Reduces Motility of HepG2 Cells via Purinergic Pathway
title_full Inhibition of ABCC6 Transporter Modifies Cytoskeleton and Reduces Motility of HepG2 Cells via Purinergic Pathway
title_fullStr Inhibition of ABCC6 Transporter Modifies Cytoskeleton and Reduces Motility of HepG2 Cells via Purinergic Pathway
title_full_unstemmed Inhibition of ABCC6 Transporter Modifies Cytoskeleton and Reduces Motility of HepG2 Cells via Purinergic Pathway
title_short Inhibition of ABCC6 Transporter Modifies Cytoskeleton and Reduces Motility of HepG2 Cells via Purinergic Pathway
title_sort inhibition of abcc6 transporter modifies cytoskeleton and reduces motility of hepg2 cells via purinergic pathway
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7349786/
https://www.ncbi.nlm.nih.gov/pubmed/32517079
http://dx.doi.org/10.3390/cells9061410
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