Detection of glutathione in oral squamous cell carcinoma cells with a fluorescent probe during the course of oxidative stress and apoptosis

Publication date: Available online 24 August 2016
Source:Journal of Oral and Maxillofacial Surgery
Author(s): Yu Zhou, Shengyun Huang, Hongzhou Shen, Meng Ma, Baocun Zhu, Dongsheng Zhang
PurposeTo detect glutathione (GSH) in oral squamous carcinoma cells (OSCCs) with a GSH selective fluorescent probe during the course of oxidative stress and apoptosis.Materials and MethodsA novel GSH probe was applied to assess GSH in human tongue squamous cell carcinoma cells cal-27. The cellular GSH and reactive oxygen species (ROS) levels were assessed with GSH probe and DCFH-DA (2,7-dichlorofuorescin diacetate) probe. The mitochondrial GSH and ROS were assessed with GSH probe, DCF-DA probe and Mitotracker Red CM-H2XRos probe. To further study whether oxidative stress would induce apoptosis of OSCCs, we then applied GSH probe and Annexin V-FITC probe to assess cellular GSH levels and eversion of phosphatidylserine, and, the cellular GSH levels and mitochondrial membrane potential (ΔΨm) were assessed with GSH probe and JC-1 probe during the course of oxidative stress and apoptosis induced by H2O2 and ethacrynic acid (EA). The fluorescence was observed under the laser confocal fluorescence microscopy.ResultsThe intensity of fluorescence which represented intracellular alteration of GSH levels, cellular ROS formation, mitochondrial ROS formation and apoptosis occurrence, respectively, could be visualized under laser confocal fluorescence microscopy.ConclusionThe GSH selective fluorescent probe can evaluate cellular GSH levels sensitively during the course of oxidative stress and apoptosis of OSCCs induced by exogenous H2O2 which could be enhanced by depletion of mtGSH.



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