Тип публикации: статья из журнала
Год издания: 2014
Идентификатор DOI: 10.1134/S1990747813050061
Ключевые слова: antitumor effect, apoptosis, DNA-aptamers, Ehrlich ascites adenocarcinoma, mass spectrometry, proliferation, protein targets of aptamers, aptamer, filamin A, phosphatidylserine, proton transporting adenosine triphosphatase, animal cell, animal tissue, article, binding affinity, cancer cell culture, cancer growth, cell cycle arrest, cell proliferation, controlled study, DNA library, Ehrlich ascites tumor cell, G1 phase cell cycle checkpoint, high performance liquid chromatography, metastasis, molecular evolution, mouse, nonhuman, oxidative phosphorylation, phospholipid transfer, priority journal, protein binding, protein DNA interaction, protein folding, protein secondary structure, protein targeting, tandem mass spectrometry
Аннотация: New prospects for the applications of single-stranded DNA and RNA as therapeutic agents have been discovered in the recent years. Aptamers are the oligonucleotides that bind to their targets with high affinity and specificity due to the well-defined tertiary structures and spatial charge distribution. Aptamers can be selected for aПоказать полностьюny molecules, virus particles, bacteria, cells, and tissues. They have a wide range of applications from target identification to drug delivery. Aptamers themselves can affect various cell functions by affecting certain proteins and receptors. Here, we present the technique for selecting aptamers with antitumor activity in cancer cell cultures and identifying their target proteins by mass spectrometry analysis. The evolved aptamers showed the following antitumor properties: AS-14 (K d = 3.8 nM) induced apoptosis (phosphatidylserine translocation determined with Annexin V Alexa Fluor 488) and AS-9 (K d = 0.75 nM) stopped proliferation (as determined with CellTrace™ Far Red DDAO-SE) in the culture of Ehrlich ascites adenocarcinoma cells. Using high performance liquid chromatography and high resolution tandem mass spectrometry, we have identified the proteins affected by the AS-14 and AS-9 aptamers. One of the most likely targets for AS-14 was filamin A, which is involved in metastasis formation, tumor development, and cell proliferation. According to mass spectrometry data, the AS-9 aptamer influences the ?-subunit of mitochondrial ATP synthase, the key component of mitochondrial oxidative phosphorylation, stimulation of which leads to tumor growth suppression. Thus, mass spectrometry data confirmed the results of the experiments on cell cultures showing that the aptamer binding to specific protein targets causes apoptosis and stops proliferation of cancer cells. However, the mechanisms of action of aptamers in vitro and in vivo are not clear enough and still need to be determined. Our study opens up new possibilities for creation of non-toxic drugs based on DNA aptamers for targeted anticancer therapy. © 2014 Pleiades Publishing, Ltd.
Журнал: Biochemistry (Moscow) Supplement Series A: Membrane and Cell Biology
Выпуск журнала: Vol. 8, Is. 1
Номера страниц: 60-72