Hübinger Lisa, Runge Roswitha, Rosenberg Tobias, Freudenberg Robert, Kotzerke Jörg, Brogsitter Claudia
Department of Nuclear Medicine, University Hospital Carl Gustav Carus, Technische Universität Dresden, 01307 Dresden, Germany.
Int J Mol Sci. 2022 Dec 3;23(23):15233. doi: 10.3390/ijms232315233.
Possible enhancements of DNA damage with light of different wavelengths and ionizing radiation (Rhenium-188-a high energy beta emitter (Re-188)) on plasmid DNA and FaDu cells via psoralen were investigated. The biophysical experimental setup could also be used to investigate additional DNA damage due to photodynamic effects, resulting from Cherenkov light. Conformational changes of plasmid DNA due to DNA damage were detected and quantified by gel electrophoresis and fluorescent staining. The clonogene survival of the FaDu cells was analyzed with colony formation assays. Dimethyl sulfoxide was chosen as a chemical modulator, and Re-188 was used to evaluate the radiotoxicity and light (UVC: = 254 nm and UVA: = 366 nm) to determine the phototoxicity. Psoralen did not show chemotoxic effects on the plasmid DNA or FaDu cells. After additional treatment with light (only 366 nm-not seen with 254 nm), a concentration-dependent increase in single strand breaks (SSBs) was visible, resulting in a decrease in the survival fraction due to the photochemical activation of psoralen. Whilst UVC light was phototoxic, UVA light did not conclude in DNA strand breaks. Re-188 showed typical radiotoxic effects with SSBs, double strand breaks, and an overall reduced cell survival for both the plasmid DNA and FaDu cells. While psoralen and UVA light showed an increased toxicity on plasmid DNA and human cancer cells, Re-188, in combination with psoralen, did not provoke additional DNA damage via Cherenkov light.
研究了通过补骨脂素,不同波长的光和电离辐射(铼 - 188 - 一种高能β发射体(Re - 188))对质粒DNA和FaDu细胞的DNA损伤的可能增强作用。该生物物理实验装置还可用于研究由切伦科夫光引起的光动力效应导致的额外DNA损伤。通过凝胶电泳和荧光染色检测并量化了由于DNA损伤导致的质粒DNA构象变化。用集落形成试验分析了FaDu细胞的克隆基因存活率。选择二甲基亚砜作为化学调节剂,并用Re - 188评估放射毒性和光(UVC:λ = 254 nm和UVA:λ = 366 nm)以确定光毒性。补骨脂素对质粒DNA或FaDu细胞未显示化学毒性作用。在用光(仅366 nm - 254 nm未见)进行额外处理后,可见单链断裂(SSB)呈浓度依赖性增加,由于补骨脂素的光化学活化导致存活分数降低。虽然UVC光具有光毒性,但UVA光并未导致DNA链断裂。Re - 188对质粒DNA和FaDu细胞均显示出典型的放射毒性作用,包括SSB、双链断裂以及总体细胞存活率降低。虽然补骨脂素和UVA光对质粒DNA和人类癌细胞显示出增加的毒性,但Re - 188与补骨脂素联合使用时,并未通过切伦科夫光引发额外的DNA损伤。