Entity of In Vitro Toxicology and Dermato-Cosmetology, Department of Pharmaceutical and Pharmacological Sciences, Vrije Universiteit Brussel, Laarbeeklaan 103, 1090 Brussels, Belgium.
Department of Pathology, School of Veterinary Medicine and Animal Science, University of São Paulo, Av. Prof. Dr. Orlando Marques de Paiva 87, Cidade Universitária, São Paulo 05508-270, Brazil.
Int J Mol Sci. 2021 Oct 29;22(21):11724. doi: 10.3390/ijms222111724.
Connexin-based channels play key roles in cellular communication and can be affected by deleterious chemicals. In this study, the effects of various genotoxic carcinogenic compounds, non-genotoxic carcinogenic compounds and non-carcinogenic compounds on the expression and functionality of connexin-based channels, both gap junctions and connexin hemichannels, were investigated in human hepatoma HepaRG cell cultures. Expression of connexin26, connexin32, and connexin43 was evaluated by means of real-time reverse transcription quantitative polymerase chain reaction analysis, immunoblot analysis and in situ immunostaining. Gap junction functionality was assessed via a scrape loading/dye transfer assay. Opening of connexin hemichannels was monitored by measuring extracellular release of adenosine triphosphate. It was found that both genotoxic and non-genotoxic carcinogenic compounds negatively affect connexin32 expression. However, no specific effects related to chemical type were observed at gap junction or connexin hemichannel functionality level.
基于连接子的通道在细胞通讯中发挥着关键作用，并且可能受到有害化学物质的影响。在这项研究中，研究了各种遗传毒性致癌化合物、非遗传毒性致癌化合物和非致癌化合物对人肝癌 HepaRG 细胞培养物中基于连接子的通道（间隙连接和连接子半通道）表达和功能的影响。通过实时逆转录定量聚合酶链反应分析、免疫印迹分析和原位免疫染色评估连接子 26、32 和 43 的表达。通过划痕加载/染料转移测定评估间隙连接功能。通过测量细胞外三磷酸腺苷的释放来监测连接子半通道的开放。结果发现，遗传毒性和非遗传毒性致癌化合物都对连接子 32 的表达产生负面影响。然而，在间隙连接或连接子半通道功能水平上，没有观察到与化学类型相关的特定影响。
