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

Cells
Angela OstuniFaustino Bisaccia

Abstract

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 cytos...Continue Reading

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Citations

Jan 21, 2021·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Fabiana GiglioAngela Ostuni
Feb 18, 2021·Scientific Reports·Christopher BramptonOlivier Le Saux
Nov 2, 2020·FEBS Letters·Shana VerschuereOlivier M Vanakker
May 1, 2021·International Journal of Molecular Sciences·Léa VeyruneNiki Christou
May 1, 2021·International Journal of Molecular Sciences·Vittorio AbruzzeseAngela Ostuni
Jan 15, 2022·Medical & Biological Engineering & Computing·Xiongshi DengLei Wang

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Methods Mentioned

BETA
transfection
PCR
electrophoresis
Fluorescence
MDA
fluorescence microscopy

Software Mentioned

GelAnalizer
ImageJ
Leica Confocal Software
Image Lab
GraphPad Prism
GraphPad
ModFit LT

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