Tracking the cellular uptake and phototoxicity of Ru(ii)-polypyridyl-1,8-naphthalimide Tröger's base conjugates†

IF 4.2 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY
Sandra A. Bright, MariaLuisa Erby, Fergus E. Poynton, Daniel Monteyne, David Pérez-Morga, Thorfinnur Gunnlaugsson, D. Clive Williams and Robert B. P. Elmes
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Abstract

Ruthenium(II) complexes are attracting significant research attention as a promising class of photosensitizers (PSs) in photodynamic therapy (PDT). Having previously reported the synthesis of two novel Ru(II)-polypyridyl-1,8-naphthalimide Tröger's base compounds 1 and 2 with interesting photophysical properties, where the emission from either the Ru(II) polypyridyl centres or the naphthalimide moieties could be used to monitor binding to nucleic acids, we sought to use these compounds to investigate further and in more detail their biological profiling, which included unravelling their mechanism of cellular uptake, cellular trafficking and cellular responses to photoexcitation. Here we demonstrate that these compounds undergo rapid time dependent uptake in HeLa cells that involved energy dependent, caveolae and lipid raft-dependent mediated endocytosis, as demonstrated by confocal imaging, and transmission and scanning electron microscopy. Following endocytosis, both compounds were shown to localise to mostly lysosomal and Golgi apparatus compartments with some accumulation in mitochondria but no localisation was found to the nucleus. Upon photoactivation, the compounds increased ROS production and induced ROS-dependent apoptotic cell death. The photo-activated compounds subsequently induced DNA damage and altered tubulin, but not actin structures, which was likely to be an indirect effect of ROS production and induced apoptosis. Furthermore, by changing the concentration of the compounds or the laser used to illuminate the cells, the mechanism of cell death could be changed from apoptosis to necrosis. This is the first detailed biological study of Ru(II)-polypyridyl Tröger's bases and clearly suggests caveolae-dependent endocytosis is responsible for cell uptake – this may also explain the lack of nuclear uptake for these compounds and similar results observed for other Ru(II)-polypyridyl complexes. These conjugates are potential candidates for further development as PDT agents and may also be useful in mechanistic studies on cell uptake and trafficking.

Abstract Image

Abstract Image

追踪 Ru(II)-Polypyridyl-1,8-naphthalimide 特罗格氏碱共轭物的细胞吸收和光毒性
钌(II)配合物作为光动力疗法(PDT)中一类前景广阔的光敏剂(PSs),正吸引着大量研究人员的关注。此前,我们曾报道合成了两种新型 Ru(II)-聚吡啶-1,8-萘二甲酰亚胺特罗格氏碱化合物 1 和 2,它们具有有趣的光物理特性,其中 Ru(II)聚吡啶中心或萘二甲酰亚胺分子的发射可用于监测与核酸的结合。在这里,我们证明了这些化合物在 HeLa 细胞中的快速摄取具有时间依赖性,涉及能量依赖性、洞穴和脂筏依赖性介导的内吞,共聚焦成像以及透射和扫描电子显微镜都证明了这一点。内吞后,这两种化合物主要定位于溶酶体和高尔基体,并在线粒体中有所积累,但在细胞核中未发现定位。光激活后,这两种化合物会增加 ROS 的产生,并诱导 ROS 依赖性细胞凋亡。光活化化合物随后会诱导 DNA 损伤并改变微管蛋白,但不会改变肌动蛋白结构,这可能是 ROS 产生和诱导细胞凋亡的间接影响。此外,通过改变化合物的浓度或照射细胞的激光,细胞死亡的机制可以从凋亡转变为坏死。这是首次对 Ru(II)- 聚吡啶特罗格氏碱基进行详细的生物学研究,清楚地表明细胞摄取是依赖于洞穴内吞作用的,这也可以解释这些化合物缺乏核摄取的原因,以及在其他 Ru(II)- 聚吡啶复合物中观察到的类似结果。这些共轭物有可能被进一步开发为 PDT 制剂,也可能有助于细胞摄取和迁移的机理研究。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
CiteScore
6.10
自引率
0.00%
发文量
128
审稿时长
10 weeks
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