Wittig Anja, Gehrke Helge, Del Favero Giorgia, Fritz Eva-Maria, Al-Rawi Marco, Diabaté Silvia, Weiss Carsten, Sami Haider, Ogris Manfred, Marko Doris
Department of Food Chemistry and Toxicology, Faculty of Chemistry, University of Vienna, Waehringer Strasse 38, 1090 Vienna, Austria.
Institute of Toxicology and Genetics (ITG), Karlsruhe Institute of Technology (KIT), Campus Nord, Hermann-von-Helmholtz-Platz 1, 76344 Eggenstein-Leopoldshafen, Germany.
Nanomaterials (Basel). 2017 Jan 13;7(1):18. doi: 10.3390/nano7010018.
Nanostructured silica particles are commonly used in biomedical and biotechnical fields, as well as, in cosmetics and food industry. Thus, their environmental and health impacts are of great interest and effects after oral uptake are only rarely investigated. In the present study, the toxicological effects of commercially available nano-scaled silica with a nominal primary diameter of 12 nm were investigated on the human gastric carcinoma cell line GXF251L. Besides the analysis of cytotoxic and proliferative effects and the comparison with effects of particles with a nominal primary diameter of 200 nm, emphasis was also given to their influence on the cellular epidermal growth factor receptor (EGFR) and mitogen-activated protein kinases (MAPK) signaling pathways-both of them deeply involved in the regulation of cellular processes like cell cycle progression, differentiation or proliferation. The investigated silica nanoparticles (NPs) were found to stimulate cell proliferation as measured by microscopy and the sulforhodamine B assay. In accordance, the nuclear level of the proliferation marker Ki-67 was enhanced in a concentration-dependent manner. At high particle concentrations also necrosis was induced. Finally, silica NPs affected the EGFR and MAPK pathways at various levels dependent on concentration and time. However, classical activation of the EGFR, to be reflected by enhanced levels of phosphorylation, could be excluded as major trigger of the proliferative stimulus. After 45 min of incubation the level of phosphorylated EGFR did not increase, whereas enhanced levels of total EGFR protein were observed. These results indicate interference with the complex homeostasis of the EGFR protein, whereby up to 24 h no impact on the transcription level was detected. In addition, downstream on the level of the MAP kinases ERK1/2 short term incubation appeared to affect total protein levels without clear increase in phosphorylation. Depending on the concentration range, enhanced levels of ERK1/2 phosphorylation were only observed after 24 h of incubation. Taken together, the present study demonstrates the potential of the tested silica particles to enhance the growth of gastric carcinoma cells. Although interference with the EGFR/MAPK cascade is observed, additional mechanisms are likely to be involved in the onset of the proliferative stimulus.
纳米结构二氧化硅颗粒常用于生物医学和生物技术领域,以及化妆品和食品工业。因此,它们对环境和健康的影响备受关注,而口服摄入后的影响却很少被研究。在本研究中,我们研究了标称初级直径为12 nm的市售纳米级二氧化硅对人胃癌细胞系GXF251L的毒理学效应。除了分析细胞毒性和增殖效应,并与标称初级直径为200 nm的颗粒的效应进行比较外,还重点研究了它们对细胞表皮生长因子受体(EGFR)和丝裂原活化蛋白激酶(MAPK)信号通路的影响——这两条信号通路都深度参与细胞周期进程、分化或增殖等细胞过程的调控。通过显微镜检查和磺基罗丹明B测定法发现,所研究的二氧化硅纳米颗粒(NPs)能刺激细胞增殖。相应地,增殖标志物Ki-67的核水平以浓度依赖的方式升高。在高颗粒浓度下也会诱导坏死。最后,二氧化硅NPs在不同水平上影响EGFR和MAPK信号通路,这取决于浓度和时间。然而,可以排除EGFR的经典激活(以磷酸化水平升高为特征)是增殖刺激的主要触发因素。孵育45分钟后,磷酸化EGFR的水平没有增加,而总EGFR蛋白水平却有所升高。这些结果表明对EGFR蛋白复杂的内稳态产生了干扰,在此过程中,长达24小时未检测到对转录水平的影响。此外,在MAP激酶ERK1/2水平的下游,短期孵育似乎会影响总蛋白水平,而磷酸化水平没有明显增加。根据浓度范围,仅在孵育24小时后才观察到ERK1/2磷酸化水平升高。综上所述,本研究证明了所测试的二氧化硅颗粒具有促进胃癌细胞生长的潜力。尽管观察到对EGFR/MAPK级联反应有干扰,但增殖刺激的起始可能还涉及其他机制。