Double-ratiometric fluorescence imaging of H2Se and O2˙− under hypoxia for exploring Na2SeO3-induced HepG2 cells' apoptosis

Abstract

Sodium selenite (Na₂SeO₃), as an anti-tumor drug for inducing tumor cells' apoptosis, has been widely studied under normoxic conditions and has been shown to exhibit oxidative stress process-induced apoptosis. However, since the real tumor environment is hypoxic, the actual mechanism is still unclear. Hence, considering the main metabolite of Na₂SeO₃ in the metabolic process to be hydrogen selenide (H₂Se) and also that it can be converted to superoxide anion (O₂˙⁻) instantaneously in the presence of oxygen, a dual-ratiometric fluorescence imaging system for simultaneous monitoring of the changes of H₂Se and O₂˙⁻ induced by Na₂SeO₃-guided tumor cell apoptosis under hypoxic conditions was constructed. Two molecular probes NIR-H₂Se and dihydroethidium were used to detect H₂Se and O₂˙⁻, respectively, whereas Rhodamine 110 was used as the reference fluorophore. Notably, H₂Se levels significantly increased under hypoxic conditions, but there was no change in the level of O₂˙⁻, which is inconsistent with the results of the previous researches. Therefore, we hypothesize that the mechanism of Na₂SeO₃-induced apoptosis for tumor cells is caused by reductive stress; also, this method can be applied for the future study of other anti-cancer selenium compounds.