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.

C VENTURA; I LEON; ASUAJE A; N ENRRIQUE; MARTIN P; M NUÑEZ; C COCCA; V MILESI

Mar del Plata

Congreso; LXIII Reunión Anual de la Sociedad Argetina de Investigaciones Clínicas (SAIC); 2018

Sociedad Argentina de Investigaciones Clínicas (SAIC)

In tumoral cells, metabolic reprogramming conduces to a large productionof acidic substances which must be extruded to avoid theintracellular acidification. We previously reported that the inhibitionof the proton channel (Hv1) by ClGBI [2-(6-chloro-1H-benzimidazol-2-yl)guanidine] differentially inhibited cell proliferation in tumorigenic(MCF-7 and MDA-MB-231) and non-tumorigenic (MCF-10A)human breast cells. Here, we show the effect of ClGBI on intracellularpH (pHi), cell viability and cell cycle of these cell lines using2D and 3D cultures. In monolayers, the pHi (BCECF ratiometric assay)and the percentage of mitotic cells (immunofluorescence usingDAPI and anti-α-tubulin antibody) were determined. The effect ofClGBI on cell viability of 3D culture MCF-7 cells (hanging drop method)was assayed by flow cytometry using propidium iodide. Results:The pHi was tested in the three cell lines after 0.5, 24 and 48h ofHv1 inhibition. Significant pHi alterations were observed only after48h of 10μM ClGBI exposure, achieving a greater acidification inMDA-MB-231 (-0.61±0.04 pH units, p