College of Veterinary Medicine, Yangzhou University, Yangzhou, Jiangsu, 225009, PR China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, Jiangsu, 225009, PR China; Joint International Research Laboratory of Agriculture and Agri-Product Safety of the Ministry of Education of China, Yangzhou University, PR China.
College of Veterinary Medicine, Yangzhou University, Yangzhou, Jiangsu, 225009, PR China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, Jiangsu, 225009, PR China; Joint International Research Laboratory of Agriculture and Agri-Product Safety of the Ministry of Education of China, Yangzhou University, PR China.
Environ Pollut. 2020 Jun;261:114149. doi: 10.1016/j.envpol.2020.114149. Epub 2020 Feb 11.
With the development of modern industry, the problem of cadmium (Cd) pollution cannot be ignored and its toxicity has caused great personal injury to humans. Poly (ADP-ribose) polymerase 1 (PARP-1) protein is a research hotspot in recent years, the research we have published shows that 5 μM of Cd-treated NRK-52E cells activated PARP-1, but the specific effects of PARP-1 on DNA damage and cell cycle is unclear. Therefore, the purpose of this study is to reveal the effect of Cd on DNA damage and cell cycle arrest in NRK-52E cells, in addition, to investigate the role of PARP-1 in mediating this effect. Western blotting, comet assay, QRT-PCR, immunofluorescence, and co-immunoprecipitation were used to detect DNA damage and cell cycle-associated protein expression. Flow cytometry was used to assess cell cycle distribution and the apoptosis rates. Results showed that after the increase in treatment time and Cd concentration, the degree of DNA damage was significantly increased, and a transition from G0/G1 to S phase arrest was observed. In addition, inhibition of PARP-1 expression exacerbated cell damage and cell cycle arrest when DNA damage was low, but attenuated cell damage and even cell cycle arrest when DNA damage was severe. These findings in this study indicate that Cd causes DNA damage in NRK-52E cells, leading to cell cycle arrest at different phases depending on the degree of DNA damage. Moreover, PARP-1 plays an important role in mediating this effect, when DNA damage is low, it functions in DNA repair, however, when DNA damage is severe, it aggravates cell damage and induces cell death.
随着现代工业的发展，镉（Cd）污染问题不容忽视，其毒性已对人类造成了严重的人身伤害。聚（ADP-核糖）聚合酶 1（PARP-1）蛋白是近年来的研究热点，我们发表的研究表明，5 μM 的 Cd 处理 NRK-52E 细胞激活了 PARP-1，但 PARP-1 对 DNA 损伤和细胞周期的具体影响尚不清楚。因此，本研究旨在揭示 Cd 对 NRK-52E 细胞中 DNA 损伤和细胞周期阻滞的影响，此外，还研究了 PARP-1 在介导这种作用中的作用。Western blot、彗星实验、QRT-PCR、免疫荧光和共免疫沉淀用于检测 DNA 损伤和细胞周期相关蛋白的表达。流式细胞术用于评估细胞周期分布和细胞凋亡率。结果表明，随着处理时间和 Cd 浓度的增加，DNA 损伤程度显著增加，并观察到从 G0/G1 期向 S 期阻滞的转变。此外，PARP-1 表达的抑制在 DNA 损伤较低时加剧了细胞损伤和细胞周期阻滞，但在 DNA 损伤严重时减弱了细胞损伤甚至细胞周期阻滞。本研究中的这些发现表明，Cd 导致 NRK-52E 细胞中的 DNA 损伤，导致细胞周期在不同阶段停滞，具体取决于 DNA 损伤的程度。此外，PARP-1 在介导这种作用中起着重要作用，当 DNA 损伤较低时，它起 DNA 修复作用，但当 DNA 损伤严重时，它会加重细胞损伤并诱导细胞死亡。
