The Evidence of HeLa Cell Apoptosis Induced with Tetraethylammonium using Proteomics and Various Analytical Methods [Molecular Bases of Disease]

December 2nd, 2013 by Huang, L., Huang, Q.-Y., Huang, H.-Q.

Tetraethylammonium (TEA) is a potassium channel (KCh) blocker applied in the functional and pharmacological studies of the KChs. The MTT assay, a colorimetric assay to quantitatively measure living cells, demonstrated that TEA reduced the HeLa cell viability dose-dependently. Flow cytometry analysis indicated an increase apoptosis rate of the HeLa cell after exposing to TEA. Patch clamp technique revealed that the K+ current of the HeLa cell was inhibited up to 80% when exposed to TEA. In addition, quantitative real-time PCR(RT-qPCR) approach set up a crosstalk among the cytotoxicity of TEA, 4-aminopyridine and anti-cancer drug such as cisplatin. Using comparative proteomics combined with MALDI-TOF MS/MS, 33 significantly changed proteins were found from TEA treatment group, among these proteins, 12 were up-regulated and 21 were down-regulated. Here, we indicated that these proteins were closely connected with many biological functions such as oxidative stress response, signal transduction, metabolism, protein synthesis and degradation. Both western-blotting and RT-qPCR approaches further verified these differential proteins. IPA software, a tool to analyze proteomic data and model biological system, was applied to analyze the interaction pathways of these proteins. The subcellular locations of the differential proteins are also predicted from Uniprot. All results above can help in our understanding of the mechanism of TEA-induced cytotoxicity and provide potential cancer biomarkers. Various experimental results in this study (like those for cisplatin) indicated that TEA is not only a KCh blocker, but also a potential anti-cancer drug.
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