Miwa Shinji, Hoffman Robert M
Department of Orthopaedic Surgery, Kanazawa University Graduate School of Medicine, Kanazawa, Japan.
AntiCancer Inc., San Diego, CA, USA.
Methods Mol Biol. 2018;1760:199-203. doi: 10.1007/978-1-4939-7745-1_18.
DNA damage repair in response to UVC irradiation was imaged in cancer cells growing in Gelfoam histoculture. UVC-induced DNA damage repair was imaged with green fluorescent protein (GFP) fused to the DNA damage response (DDR)-related binding protein 53BP1 in MiaPaCa-2 human pancreatic cancer cells. Three-dimensional Gelfoam histocultures and confocal imaging enabled 53BP1-GFP nuclear foci to be observed within 1 h after UVC irradiation, indicating the onset of DNA damage repair response. Induction of UV-induced 53BP1-GFP focus formation was limited up to a depth of 40 μm in Gelfoam histoculture of MiaPaCa-2 cells, indicating this was the depth limit of UVC irradiation.
在Gelfoam组织培养中生长的癌细胞中，对紫外线C（UVC）照射后的DNA损伤修复进行了成像。在MiaPaCa-2人胰腺癌细胞中，用与DNA损伤反应（DDR）相关的结合蛋白53BP1融合的绿色荧光蛋白（GFP）对UVC诱导的DNA损伤修复进行成像。三维Gelfoam组织培养和共聚焦成像能够在UVC照射后1小时内观察到53BP1-GFP核灶，表明DNA损伤修复反应开始。在MiaPaCa-2细胞的Gelfoam组织培养中，紫外线诱导的53BP1-GFP灶形成的诱导在深度达40μm时受到限制，表明这是UVC照射的深度极限。
