Department of Life Sciences, College of Health, Medicine and Life Sciences, Brunel University London, Kingston Lane, Uxbridge UB8 3PH, UK.
Department of Life Sciences, College of Health, Medicine and Life Sciences, Brunel University London, Kingston Lane, Uxbridge UB8 3PH, UK.
Mutat Res Genet Toxicol Environ Mutagen. 2022 Jun;878:503476. doi: 10.1016/j.mrgentox.2022.503476. Epub 2022 Feb 24.
We examined frequencies of radiation-induced chromosomal aberrations, using classical cytological methods, and DNA damage in interphase and metaphase cells, using a combination of FISH, CO-FISH, TIF (telomere dysfunction induced assay) and simultaneous detection of DNA damage and telomeric sequences in metaphase chromosomes, in Chinese hamster cells defective in BRCA2 and control cells. Given that the Chinese hamster genome contains large blocks of interstitial telomeric sites, our results allow us to examine the role of BRCA2 in the potential fragility of these sites, but also whether BRCA2 affects DNA repair within terminal telomeric sequences. BRCA2 defective cells exhibited greater frequencies of DNA damage within interstitial telomeric sites, as well as within terminal telomeric sites, relative to control cells. Therefore, BRCA2 deficiency contributes to the telomere dysfunction phenotype in Chinese hamster cells.
我们使用经典的细胞遗传学方法检测了染色体辐射诱导的畸变频率，并用 FISH、CO-FISH、TIF（端粒功能障碍诱导分析）以及同时检测中期染色体中 DNA 损伤和端粒序列的方法检测了间期和中期细胞中的 DNA 损伤，这些实验都是在 BRCA2 缺陷的中国仓鼠细胞和对照细胞中进行的。由于中国仓鼠基因组中含有大量的染色体间端粒位点，我们的结果不仅允许我们研究 BRCA2 在这些位点潜在不稳定性中的作用，还研究了 BRCA2 是否影响端粒序列内的 DNA 修复。与对照细胞相比，BRCA2 缺陷细胞在染色体间端粒位点和端粒末端的 DNA 损伤频率更高。因此，BRCA2 缺陷导致中国仓鼠细胞中端粒功能障碍表型。
