Ujvarosi Kinga, Hunyadi Janos, Nagy Gabor, Pocsi Imre, Banfalvi Gaspar
Department of Microbial Biotechnology and Cell Biology, University of Debrecen, 1 Egyetem Square, Debrecen 4010, Hungary.
Apoptosis. 2007 Nov;12(11):2089-99. doi: 10.1007/s10495-007-0118-0. Epub 2007 Aug 14.
Exponentially growing human erythroleukemia K562 cells were permeabilized and the dose dependent decrease of DNA synthesis rate was measured after ultraviolet (UV B, 290 nm) irradiation. Cells were able to overcome 2 and 5 J/m2 UV doses, partial recovery was observed at 15 J/m2, while at high (25 J/m2) UV dose replicative DNA synthesis remained suppressed. K562 cells were subjected to synchronization prior to and after UV irradiation (24 J/m2) and 18 fractions were collected by centrifugal elutriation. Cell cycle analysis by flow cytometry did not show early apoptotic cells after UV irradiation. The gradual increase in DNA content typical for non-irradiated cells was contrasted by an early S phase block between 2.2 and 2.4 C-values after UV irradiation. Cell cycle dependent chromatin changes after ultraviolet irradiation were seen as a fine fibrillary network covering the mainly fibrous chromatin structures and incompletely folded primitive chromosomes. Based on observations after UV irradiation and on earlier results with cadmium treatment and gamma irradiation, we confirm that typical chromatin changes characteristic to genotoxic agents can be recognized and classified.
对数生长期的人红白血病K562细胞经通透处理后,测定紫外线(UV B,290 nm)照射后DNA合成速率的剂量依赖性降低。细胞能够耐受2 J/m²和5 J/m²的紫外线剂量,在15 J/m²时观察到部分恢复,而在高剂量(25 J/m²)紫外线照射下,复制性DNA合成仍受到抑制。在紫外线照射(24 J/m²)前后对K562细胞进行同步化处理,并通过离心淘析收集18个组分。流式细胞术进行的细胞周期分析未显示紫外线照射后有早期凋亡细胞。紫外线照射后,未照射细胞典型的DNA含量逐渐增加与早期S期阻滞形成对比,紫外线照射后在2.2至2.4 C值之间出现早期S期阻滞。紫外线照射后细胞周期依赖性染色质变化表现为覆盖主要为纤维状染色质结构和未完全折叠的原始染色体的精细纤维网络。基于紫外线照射后的观察结果以及镉处理和γ射线照射的早期结果,我们证实可以识别和分类遗传毒性剂特有的典型染色质变化。