Differential effects of HV1 proton channel inhibition on intracellular pH and cell cycle progression of tumorigenic and non-tumorigenic human breast cells. 2d and 3d studies.

ASUAJE, AGUSTÍN; NUÑEZ, MARIEL; VENTURA, CLARA; ENRIQUE, NICOLÁS; COCCA, CLAUDIA; LEÓN, IGNACIO E.; MARTÍN, PEDRO; MILESI, VERÓNICA

Mar del Plata

Congreso; Reunion Conjunta SAIC SAI SAFIS 2018; 2018

SAIC SAI SAFIS

Metabolic reprogramming conduces to a large production of acidic substances which must be extruded of the cells. We have previously reported that the inhibition of the proton channel Hv1 by ClGBI [2-(6-chloro-1H-benzimidazol-2-yl)guanidine] differentially affect cell proliferation in tumorigenic (MCF-7 and MDA-MB-231) and non-tumorigenic (MCF-10A) human breast cell lines. Here, we studied the effect of ClGBI on intracellular pH (pHi), cell viability and cell cycle of tumorigenic and non-tumorigenic human breast cell lines using 2D and 3D cultures. In monolayers, pHi (BCECF ratiometric assay) and the percentage of mitotic cells (immunofluorescence using DAPI and anti-α-tubulin antibody) were determined. The effect of ClGBI on cell viability of MCF-7 cells in 3D culture (hanging drop method) was assayed by flow cytometry using propidium iodide. Results: pHi was differentially affected in the three cell lines after 0.5, 24 and 48h of Hv1 inhibition. Significant pHi alterations were observed only after 48h of 10μM ClGBI exposure, achieving a greater acidification in MDA-MB-231 (-0.61±0.04 pH units, p