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Proliferation assay (MTT)

MN María del Mar Noblejas-López
CN Cristina Nieto-Jiménez
EG Eva M. Galán-Moya
DT David Tebar-García
JM Juan Carlos Montero
AP Atanasio Pandiella
MB Miguel Burgos
AO Alberto Ocaña
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Anti-proliferative effect was evaluated through MTT (Sigma Aldrich) colorimetric assay. Cells were seeded on 48-wells plates and were treated 24 h later with indicated doses of each drug, alone or in combination. Cell medium was replaced at 72 h with MTT solution (red phenol-free DMEM with MTT 0.5 μg/μL) for 45 min at 37 °C. DMSO was then used to solubilize the samples. Absorbances at 555 nm values were recorded in a spectrophotometer multiwell plate reader. A reference wavelength of 690 nm was used.

To analyze if the effect of the compounds was additive or synergic, we combined different doses of MZ1 and trastuzumab and we used the Calcusyn 2.0 software (Biosoft). The mathematical expression used is the Chou-Talalay algorithm, which allows to obtain the combination index (CI) indicating synergistic effect (< 1), additive effect (=1) and antagonistic effect (> 1).

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