Di Somma Sarah, Amato Jussara, Iaccarino Nunzia, Pagano Bruno, Randazzo Antonio, Portella Giuseppe, Malfitano Anna Maria
Department of Translational Medical Sciences, University of Naples Federico II, 80131 Naples, Italy.
Department of Pharmacy, University of Naples Federico II, 80131 Naples, Italy.
Cancers (Basel). 2019 Nov 15;11(11):1797. doi: 10.3390/cancers11111797.
DNA G-quadruplex (G4) structures represent potential anti-cancer targets. In this study, we compared the effect of two G4-targeting compounds, C066-3108 and the gold standard BRACO-19.
In breast and prostate cancer cells, cytotoxicity induced by both molecules was measured by a sulforhodamine B assay. In breast cancer cells, cycle, apoptosis, the formation of G4 structures, calreticulin and high mobility group box 1 (HMGB1), as well as T cell activation, were analyzed by flow cytometry and adenosine triphosphate (ATP) by luminescence.
Both ligands inhibited cell survival and induced DNA damage. In MCF-7 cells, G4 ligands increased the subG0/G1 phase of the cell cycle inducing apoptosis and reduced intracellular ATP. In untreated MCF-7 cells, we observed a slight presence of G4 structures associated with the G2/M phase. In MDA-MB231 cells, G4 ligands decreased the G1 and enhanced the G2/M phase. We observed a decrease of intracellular ATP, calreticulin cell surface exposure and an increase of HMGB1, accompanied by T cell activation. Both compounds induced G4 structure formation in the subG0/G1 phase.
Our data report similar effects for both compounds and the first evidence that G4 ligands induce the release of danger signals associated with immunogenic cell death and induction of T cell activation.
DNA G-四链体（G4）结构是潜在的抗癌靶点。在本研究中，我们比较了两种靶向G4的化合物C066-3108和金标准化合物BRACO-19的作用效果。
在乳腺癌和前列腺癌细胞中，通过磺酰罗丹明B测定法检测两种分子诱导的细胞毒性。在乳腺癌细胞中，通过流式细胞术分析细胞周期、凋亡、G4结构的形成、钙网蛋白和高迁移率族蛋白B1（HMGB1），并通过发光法检测三磷酸腺苷（ATP）。
两种配体均抑制细胞存活并诱导DNA损伤。在MCF-7细胞中，G4配体增加了细胞周期的亚G0/G1期，诱导凋亡并降低细胞内ATP。在未处理的MCF-7细胞中，我们观察到与G2/M期相关的G4结构略有存在。在MDA-MB231细胞中，G4配体减少了G1期并增强了G2/M期。我们观察到细胞内ATP减少、钙网蛋白细胞表面暴露减少以及HMGB1增加，同时伴有T细胞活化。两种化合物均在亚G0/G1期诱导G4结构形成。
我们的数据报告了两种化合物的相似作用效果，并且首次证明G4配体诱导与免疫原性细胞死亡相关的危险信号释放并诱导T细胞活化。
