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.

VENTURA C; LEON I; ASUAJE A; ENRIQUE N; MARTIN P; NUÑEZ M; COCCA C

Congreso; LXIII Reunión Científica Anual de la Sociedad Argentina de Investigación Clínica (SAIC); 2018

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] affect cell proliferation in tumorigenic (MCF-7 and MDA-MB-231) and non-tumorigenic (MCF-10A) human breast cell lines, differentially. Here, we show 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, the 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: The 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