Yurova M N, Fedoros E I, Danilova A B, Nekhaeva T L, Emelyanova N V, Grigorevskaya A V, Popovich I G, Baldueva I A
N. N. Petrov National Medical Research Center of Oncology, Ministry of Health of Russian Federation, St. Petersburg, Russia.
Bull Exp Biol Med. 2025 Aug 28. doi: 10.1007/s10517-025-06465-0.
The aim was to test the protocol for obtaining lymphoid tissue from mouse lymph nodes and characterize the level of DNA damage with analysis of the repair and apoptosis protein markers in ex vivo model of cell irradiation. The lymph node cell suspension was exposed to X-ray irradiation at a dose of 2 Gy and compared with a sham-irradiated (0 Gy) control. Ex vivo irradiation at a dose of 2 Gy after 3 h caused a 7-fold increase in DNA damage level determined in DNA comet assay. A significant increase in the level of repair proteins γH2AX (by 3.7 times), Rad51 (by 2.2 times), and XPA (by 28 times), cell cycle arrest proteins Cdc25A (by 2.6 times), p21 (by 10 times), and p16 (by 5 times), and pro-apoptotic protein Bax (by 3.8 times) was noted, without significant changes in Bcl-2 level. The proposed technique allows quick obtaining lymphoid tissue samples suitable for analysis of the radiosensitivity of lymphoid cells in mice of various strains, changes in their sensitivity after or in combination with the primary damaging exposure.
目的是测试从小鼠淋巴结获取淋巴组织的方案，并通过分析细胞照射体外模型中的修复和凋亡蛋白标志物来表征DNA损伤水平。将淋巴结细胞悬液暴露于2 Gy的X射线照射下，并与假照射（0 Gy）对照进行比较。3小时后2 Gy剂量的体外照射导致DNA彗星试验中测定的DNA损伤水平增加了7倍。观察到修复蛋白γH2AX（增加3.7倍）、Rad51（增加2.2倍）和XPA（增加28倍）、细胞周期阻滞蛋白Cdc25A（增加2.6倍）、p21（增加10倍）和p16（增加5倍）以及促凋亡蛋白Bax（增加3.8倍）的水平显著增加，而Bcl-2水平无显著变化。所提出的技术能够快速获取适合分析各种品系小鼠淋巴细胞放射敏感性、其在初次损伤暴露后或与之联合时敏感性变化的淋巴组织样本。
