Zinovkina Ludmila A, Galivondzhyan Alina K, Prikhodko Anastasia S, Galkin Ivan I, Zinovkin Roman A
Faculty of Bioengineering and Bioinformatics, Lomonosov Moscow State University, Moscow, Russia.
Institute of Mitoengineering, Moscow State University, Moscow, Russia.
PeerJ. 2020 Mar 25;8:e8803. doi: 10.7717/peerj.8803. eCollection 2020.
Targeting negatively charged mitochondria is often achieved using triphenylphosphonium (TPP) cations. These cationic vehicles may possess biological activity, and a docking study indicates that TPP-moieties may act as modulators of signaling through the estrogen receptor α (ERα). Moreover, in vivo and in vitro experiments revealed the estrogen-like effects of TPP-based compounds. Here, we tested the hypothesis that TPP-based compounds regulate the activity of ERα.
We used ERa-positive and ERα-negative human breast adenocarcinoma cell lines (MCF-7 and MDA-MB-231, respectively). Cell proliferation was measured using a resazurin cell growth assay and a real-time cell analyzer assay. Cell cycle progression was analyzed using flow cytometry. Real-time PCR was used to assess mRNA expression of endogenous estrogen-responsive genes. Luciferase activity was measured to evaluate transcription driven by estrogen-responsive promoters in cells transfected with an estrogen response element (ERE)-luciferase expression vector.
The TPP-based molecules SkQ1 and CTPP, as well as the rhodamine-based SkQR1, did not increase the proliferation or alter the cell cycle progression of MCF-7 cells. In contrast, 17β estradiol increased the proliferation of MCF-7 cells and the proportion of cells in the S/G2/M-phases of the cell cycle. TPP-based compounds did not affect the induction of transcription of an ERE-luciferase expression vector in vitro, and SkQ1 did not alter the levels of expression of estrogen-dependent genes encoding GREB1, TFF1, COX6, and IGFBP4.
TPP-based compounds do not possess properties typical of ERα agonists.
使用三苯基膦（TPP）阳离子通常可实现对带负电荷线粒体的靶向作用。这些阳离子载体可能具有生物活性，一项对接研究表明，TPP部分可能作为雌激素受体α（ERα）信号传导的调节剂。此外，体内和体外实验揭示了基于TPP的化合物具有雌激素样作用。在此，我们检验了基于TPP的化合物调节ERα活性这一假设。
我们使用了ERα阳性和ERα阴性的人乳腺癌细胞系（分别为MCF-7和MDA-MB-231）。使用刃天青细胞生长测定法和实时细胞分析仪测定法测量细胞增殖。使用流式细胞术分析细胞周期进程。使用实时PCR评估内源性雌激素反应基因的mRNA表达。在转染了雌激素反应元件（ERE）-荧光素酶表达载体的细胞中，测量荧光素酶活性以评估由雌激素反应启动子驱动的转录。
基于TPP的分子SkQ1和CTPP，以及基于罗丹明的SkQR1，均未增加MCF-7细胞的增殖或改变其细胞周期进程。相比之下，17β雌二醇增加了MCF-7细胞的增殖以及细胞周期S/G2/M期的细胞比例。基于TPP的化合物在体外不影响ERE-荧光素酶表达载体的转录诱导，并且SkQ1不改变编码GREB1、TFF1、COX6和IGFBP4的雌激素依赖性基因的表达水平。
基于TPP的化合物不具有ERα激动剂的典型特性。
