title: The evidence of HeLa cell apoptosis induced with tetraethylammonium using proteomics and various analytical methods.
abstract: Tetraethylammonium (TEA) is a potassium channel (KCh) blocker applied in the functional and pharmacological studies of the KChs. The MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, a colorimetric assay to quantitatively measure living cells, demonstrated that TEA reduced the HeLa cell viability dose-dependently. Flow cytometry analysis indicated an increased apoptosis rate of the HeLa cell after exposing to TEA. The 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 approach set up cross-talk 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 quantitative real-time PCR approaches further verified these differential proteins. Ingenuity Pathways Analysis software, a tool to analyze "omics" 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.
authors.affiliation: From the State Key Laboratory of Cellular Stress Biology, School of Life Sciences, State Key Laboratory of Marine Environmental Science, Xiamen University, Xiamen 361005, China and.
authors.forename: Lin
authors.initials: L
authors.lastname: Huang
authors.forename: Qing-Yu
authors.initials: QY
authors.lastname: Huang
authors.forename: He-Qing
authors.initials: HQ
authors.lastname: Huang
mesh_terms: D015761:4-Aminopyridine / Q000494:pharmacology; D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D002945:Cisplatin / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D006367:HeLa Cells; D006801:Humans; D011188:Potassium / Q000378:metabolism*; D026902:Potassium Channel Blockers / Q000494:pharmacology*; D020543:Proteome / Q000096:biosynthesis*; D040901:Proteomics; D060888:Real-Time Polymerase Chain Reaction; D019789:Tetraethylammonium / Q000494:pharmacology*
chemical_list: D000970:Antineoplastic Agents; D026902:Potassium Channel Blockers; D020543:Proteome; D019789:Tetraethylammonium; D015761:4-Aminopyridine; D002945:Cisplatin; D011188:Potassium
keywords: Apoptosis; Bioinformatics; Flow Cytometry; HeLa Cell Line; Patch Clamp Technology; Pharmacology; Proteomics; RT-qPCR; Tetraethylammonium; Western Blotting