The proton channel Hv1 inhibition induces differential effects on tumorigenic (2D and 3D) and non-tumorigenic human breast cells.

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

Bratislava

Congreso; 5th Annual Meeting ? Metabolic Adaptations and Targets in Cancer ?; 2018

International Society of Cancer Metabolism

Metabolic reprogramming of cancer cells conduces to a high production of acidic substances that must be extruded to maintain cell viability. In this work, we studied the effect of an inhibitor of the proton channel HVCN1 [2-(6-chloro-1H-benzimidazol-2-yl)guanidine (ClGBI)], on intracellular pH (pHi), proliferation and cell cycle of tumorigenic (MDA-MB-231 and MCF-7) and non-tumorigenic (MCF-10A) human breast cell lines in 2D culture. Concentration-response curve and time-course experiments were assayed to study pHi (by BCEF ratiometric method) cell proliferation (clonogenic assay), cell viability (MTT), cell cycle distribution (flow cytometry) and the percentage of mitotic cells (immunofluorescence using DAPI and anti-α-tubulin antibody). Additionally, the effect of ClGBI was also assayed on cell viability of tumorigenic cells in 3D culture of MCF-7 cells (hanging drop method) by flow cytometry using propidium iodide as a viability marker. Results: First, we tested if ClGBI was able to modify the basal pHi of the three cell lines, observing that 10 µM ClGBI did not affect the MCF-10A but significantly induces acidification in tumorigenic cells MCF-7 and MDA-MB-231 (a decrease of 0.06±0.003 and 0.16±0.036 pHi units, respectively p