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Revisiting CFTR inhibition: a comparative study of CFTRinh -172 and GlyH-101 inhibitors.

Melis N, Tauc M, Cougnon M, Bendahhou S, Giuliano S, Rubera I, Duranton C - Br. J. Pharmacol. (2014)

Bottom Line: We also explored the effect of both inhibitors on cell viability using live/dead and cell proliferation assays in two different cell lines.The CFTRinh -172 did not affect the CaCC but did inhibit the VSORC, at concentrations higher than 5 µM.Our results provided insights into their use in mouse models.

View Article: PubMed Central - PubMed

Affiliation: University of Nice-Sophia Antipolis, LP2M CNRS-UMR7370, Faculté de médecine, Nice, France.

No MeSH data available.


Related in: MedlinePlus

Effect of CFTRinh-172 and GlyH-101 inhibitors on cellular toxicity. (A and B). Representative fluorescent dye staining (A) and related quantification of cell death (B) in CFTR-expressing cells (confluent kidney PCT cell monolayers) exposed for 24 h to increasing concentrations of CFTRinh-172 or GlyH-101 (concentrations ranging from 0.5 to 50 µM). Live cells labelled with calcein-AM appeared green while dead cells labelled with homodimeric propidium iodide appeared red. Scale bars represents 80 µm. Values were normalized to the 100% of live cells in control experiments and were means (±SEM) of three to six individual experiments. (C) MTT assay performed on CFTR-expressing cells (kidney PCT cells) exposed as in (A) to increasing concentrations of both CFTR inhibitors. Values were normalized to control experiments and represent means (±SEM) of eight individual experiments. (D) MTT assay performed on non–CFTR-expressing cells (PS120) exposed to increasing concentrations of both inhibitors. Values were normalized to vehicle experiments and represent means (SEM) of eight individual experiments. *P < 0.05, Tukey's HSD test.
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fig01: Effect of CFTRinh-172 and GlyH-101 inhibitors on cellular toxicity. (A and B). Representative fluorescent dye staining (A) and related quantification of cell death (B) in CFTR-expressing cells (confluent kidney PCT cell monolayers) exposed for 24 h to increasing concentrations of CFTRinh-172 or GlyH-101 (concentrations ranging from 0.5 to 50 µM). Live cells labelled with calcein-AM appeared green while dead cells labelled with homodimeric propidium iodide appeared red. Scale bars represents 80 µm. Values were normalized to the 100% of live cells in control experiments and were means (±SEM) of three to six individual experiments. (C) MTT assay performed on CFTR-expressing cells (kidney PCT cells) exposed as in (A) to increasing concentrations of both CFTR inhibitors. Values were normalized to control experiments and represent means (±SEM) of eight individual experiments. (D) MTT assay performed on non–CFTR-expressing cells (PS120) exposed to increasing concentrations of both inhibitors. Values were normalized to vehicle experiments and represent means (SEM) of eight individual experiments. *P < 0.05, Tukey's HSD test.

Mentions: We first evaluated the cytotoxicity of CFTRinh-172 and GlyH-101 in kidney cells. Confluent cell cultures were exposed to increasing concentrations of GlyH-101 and CFTRinh-172 (1–50 µM), and the fluorescent dye assay used to determine cytotoxicity (live/dead labelling) after incubation for 24 h. Figure 1A illustrates confluent cell monolayers in the absence and presence of CFTRinh-172 (upper part) or GlyH-101 (lower part). From 1 to 20 µM, CFTRinh-172 and GlyH-101 had no significant effect on cell viability as indicated by a homogeneous green labeling of the monolayers (Figure 1A and B). However at a higher concentration (50 µM), both substances induced cell death as revealed by the decrease of green-labelled areas and a simultaneous increase in red positive cells (dead cells). To confirm the toxicity of both substances, we performed MTT assays on confluent CFTR-expressing cells monolayers. These experiments confirmed for both substances a dramatic decrease of the cell viability at 50 µM but revealed also a marked effect for lower concentrations (20 µM and even 10 µM, Figure 1C). MTT assays were also performed on non–CFTR-expressing PS120 cells. GlyH-101 also decreased cell viability (Figure 1D) at concentrations higher than 5 µM. Interestingly, CFTRinh-172 was more cytotoxic than GlyH-101 and exhibited a significant effect at 5 µM.


Revisiting CFTR inhibition: a comparative study of CFTRinh -172 and GlyH-101 inhibitors.

Melis N, Tauc M, Cougnon M, Bendahhou S, Giuliano S, Rubera I, Duranton C - Br. J. Pharmacol. (2014)

Effect of CFTRinh-172 and GlyH-101 inhibitors on cellular toxicity. (A and B). Representative fluorescent dye staining (A) and related quantification of cell death (B) in CFTR-expressing cells (confluent kidney PCT cell monolayers) exposed for 24 h to increasing concentrations of CFTRinh-172 or GlyH-101 (concentrations ranging from 0.5 to 50 µM). Live cells labelled with calcein-AM appeared green while dead cells labelled with homodimeric propidium iodide appeared red. Scale bars represents 80 µm. Values were normalized to the 100% of live cells in control experiments and were means (±SEM) of three to six individual experiments. (C) MTT assay performed on CFTR-expressing cells (kidney PCT cells) exposed as in (A) to increasing concentrations of both CFTR inhibitors. Values were normalized to control experiments and represent means (±SEM) of eight individual experiments. (D) MTT assay performed on non–CFTR-expressing cells (PS120) exposed to increasing concentrations of both inhibitors. Values were normalized to vehicle experiments and represent means (SEM) of eight individual experiments. *P < 0.05, Tukey's HSD test.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4128068&req=5

fig01: Effect of CFTRinh-172 and GlyH-101 inhibitors on cellular toxicity. (A and B). Representative fluorescent dye staining (A) and related quantification of cell death (B) in CFTR-expressing cells (confluent kidney PCT cell monolayers) exposed for 24 h to increasing concentrations of CFTRinh-172 or GlyH-101 (concentrations ranging from 0.5 to 50 µM). Live cells labelled with calcein-AM appeared green while dead cells labelled with homodimeric propidium iodide appeared red. Scale bars represents 80 µm. Values were normalized to the 100% of live cells in control experiments and were means (±SEM) of three to six individual experiments. (C) MTT assay performed on CFTR-expressing cells (kidney PCT cells) exposed as in (A) to increasing concentrations of both CFTR inhibitors. Values were normalized to control experiments and represent means (±SEM) of eight individual experiments. (D) MTT assay performed on non–CFTR-expressing cells (PS120) exposed to increasing concentrations of both inhibitors. Values were normalized to vehicle experiments and represent means (SEM) of eight individual experiments. *P < 0.05, Tukey's HSD test.
Mentions: We first evaluated the cytotoxicity of CFTRinh-172 and GlyH-101 in kidney cells. Confluent cell cultures were exposed to increasing concentrations of GlyH-101 and CFTRinh-172 (1–50 µM), and the fluorescent dye assay used to determine cytotoxicity (live/dead labelling) after incubation for 24 h. Figure 1A illustrates confluent cell monolayers in the absence and presence of CFTRinh-172 (upper part) or GlyH-101 (lower part). From 1 to 20 µM, CFTRinh-172 and GlyH-101 had no significant effect on cell viability as indicated by a homogeneous green labeling of the monolayers (Figure 1A and B). However at a higher concentration (50 µM), both substances induced cell death as revealed by the decrease of green-labelled areas and a simultaneous increase in red positive cells (dead cells). To confirm the toxicity of both substances, we performed MTT assays on confluent CFTR-expressing cells monolayers. These experiments confirmed for both substances a dramatic decrease of the cell viability at 50 µM but revealed also a marked effect for lower concentrations (20 µM and even 10 µM, Figure 1C). MTT assays were also performed on non–CFTR-expressing PS120 cells. GlyH-101 also decreased cell viability (Figure 1D) at concentrations higher than 5 µM. Interestingly, CFTRinh-172 was more cytotoxic than GlyH-101 and exhibited a significant effect at 5 µM.

Bottom Line: We also explored the effect of both inhibitors on cell viability using live/dead and cell proliferation assays in two different cell lines.The CFTRinh -172 did not affect the CaCC but did inhibit the VSORC, at concentrations higher than 5 µM.Our results provided insights into their use in mouse models.

View Article: PubMed Central - PubMed

Affiliation: University of Nice-Sophia Antipolis, LP2M CNRS-UMR7370, Faculté de médecine, Nice, France.

No MeSH data available.


Related in: MedlinePlus