Nickel nanoparticle-induced cell transformation: involvement of DNA damage and DNA repair defect through HIF-1α/miR-210/Rad52 pathway.
Nickel nanoparticle-induced cell transformation: involvement of DNA damage and DNA repair defect through HIF-1α/miR-210/Rad52 pathway.
复制标题
DOI：
10.1186/s12951-021-01117-7
复制
发表时间：
2021-11-17
影响因子：
10.2
通讯作者：
Zhang Q
中科院分区：
文献类型：
--
作者：
Mo Y;Zhang Y;Zhang Y;Yuan J;Mo L;Zhang Q
随着纳米技术的发展，纳米镍在工业和生物医学中的应用越来越广泛。然而，纳米镍的遗传毒性和致癌作用及其潜在机制尚不清楚。首先，采用real-time PCR或Western blot方法，对永生化人支气管上皮细胞BEAS-2B进行剂量反应（0、10、20和30 μg/mL）和时间反应（0、3、6、12和24 h）研究，观察纳米ni对DNA损伤反应（DDR）相关蛋白和HIF-1α/miR-210/Rad52通路的影响。然后，使用Hsp90抑制剂（1µM 17-AAG，间接HIF-1α抑制剂）、HIF-1α敲除（KO）细胞和miR-210抑制剂（20 nM）来确定纳米ni诱导的Rad52下调是否通过HIF-1α核积累和miR-210上调来实现。在长期实验中，分别用0.25和0.5µg/mL的纳米ni处理细胞21个周期（~ 150天），并通过软琼脂培养检测细胞的非锚定生长水平。将含有人Rad52 ORF的慢病毒颗粒转导到BEAS-2B细胞中，观察Rad52在纳米镍诱导细胞转化中的作用。通过气管内滴入50µg纳米ni，研究了纳米ni诱导的DNA损伤和HIF-1α/miR-210/Rad52通路的失调。采用gpt delta转基因小鼠分析纳米ni暴露后小鼠肺内突变频率和突变谱。纳米镍暴露在体外和体内都会引起DNA损伤，这反映在ddr相关蛋白的磷酸化增加，如Ser1981位点的ATM、Ser15位点的p53和H2AX。纳米ni暴露还诱导HIF-1α核积累，miR-210上调，同源重组修复（HRR）基因Rad52下调。抑制或敲除HIF-1α或miR-210可改善纳米ni诱导的Rad52下调。长期低剂量纳米镍暴露可导致细胞恶性转化，Rad52表达增加可显著降低纳米镍诱导的细胞转化。此外，在纳米镍暴露后第7天和第42天，小鼠肺细支气管上皮细胞和增生性肺细胞中细胞增殖标志物Ki-67和PCNA的免疫染色增加。最后，使用gpt delta转基因小鼠发现，纳米ni暴露不会导致gpt突变频率增加，某些DNA突变，如碱基置换和小碱基插入/缺失，并不是纳米ni诱导的DNA损伤的主要类型。本研究揭示了纳米镍诱导细胞恶性转化的机制；纳米镍诱导的DNA损伤和通过HIF-1α/miR-210/Rad52途径的DNA修复缺陷的共同作用可能导致纳米镍诱导的基因组不稳定并最终导致细胞转化。本研究结果将为进一步阐明纳米镍致遗传毒性和致癌性的分子机制提供信息。在线版本包含补充材料，可在10.1186/s12951-021-01117-7获得。
Nickel nanoparticles (Nano-Ni) are increasingly used in industry and biomedicine with the development of nanotechnology. However, the genotoxic and carcinogenic effects of Nano-Ni and the underlying mechanisms are still unclear. At first, dose–response (0, 10, 20, and 30 μg/mL) and time-response (0, 3, 6, 12, and 24 h) studies were performed in immortalized normal human bronchial epithelial cells BEAS-2B to observe the effects of Nano-Ni on DNA damage response (DDR)-associated proteins and the HIF-1α/miR-210/Rad52 pathway by real-time PCR or Western blot. Then, a Hsp90 inhibitor (1 µM of 17-AAG, an indirect HIF-1α inhibitor), HIF-1α knock-out (KO) cells, and a miR-210 inhibitor (20 nM) were used to determine whether Nano-Ni-induced Rad52 down-regulation was through HIF-1α nuclear accumulation and miR-210 up-regulation. In the long-term experiments, cells were treated with 0.25 and 0.5 µg/mL of Nano-Ni for 21 cycles (~ 150 days), and the level of anchorage-independent growth was determined by plating the cells in soft agar. Transduction of lentiviral particles containing human Rad52 ORF into BEAS-2B cells was used to observe the role of Rad52 in Nano-Ni-induced cell transformation. Nano-Ni-induced DNA damage and dysregulation of HIF-1α/miR-210/Rad52 pathway were also investigated in vivo by intratracheal instillation of 50 µg per mouse of Nano-Ni. gpt delta transgenic mice were used to analyze mutant frequency and mutation spectrum in mouse lungs after Nano-Ni exposure. Nano-Ni exposure caused DNA damage at both in vitro and in vivo settings, which was reflected by increased phosphorylation of DDR-associated proteins such as ATM at Ser1981, p53 at Ser15, and H2AX. Nano-Ni exposure also induced HIF-1α nuclear accumulation, miR-210 up-regulation, and down-regulation of homologous recombination repair (HRR) gene Rad52. Inhibition of or knocking-out HIF-1α or miR-210 ameliorated Nano-Ni-induced Rad52 down-regulation. Long-term low-dose Nano-Ni exposure led to cell malignant transformation, and augmentation of Rad52 expression significantly reduced Nano-Ni-induced cell transformation. In addition, increased immunostaining of cell proliferation markers, Ki-67 and PCNA, was observed in bronchiolar epithelial cells and hyperplastic pneumocytes in mouse lungs at day 7 and day 42 after Nano-Ni exposure. Finally, using gpt delta transgenic mice revealed that Nano-Ni exposure did not cause increased gpt mutant frequency and certain DNA mutations, such as base substitution and small base insertions/deletions, are not the main types of Nano-Ni-induced DNA damage. This study unraveled the mechanisms underlying Nano-Ni-induced cell malignant transformation; the combined effects of Nano-Ni-induced DNA damage and DNA repair defects through HIF-1α/miR-210/Rad52 pathway likely contribute to Nano-Ni-induced genomic instability and ultimately cell transformation. Our findings will provide information to further elucidate the molecular mechanisms of Nano-Ni-induced genotoxicity and carcinogenicity. The online version contains supplementary material available at 10.1186/s12951-021-01117-7.
登录
查看更多内容
影响因子：
2.8
作者：
Åkerlund E;Cappellini F;Di Bucchianico S;Islam S;Skoglund S;Derr R;Odnevall Wallinder I;Hendriks G;Karlsson HL
通讯作者：
Karlsson HL
DOI：
10.1016/bs.enz.2016.03.003
发表时间：
2016
期刊：
The Enzymes
影响因子：
--
作者：
Boehm EM;Gildenberg MS;Washington MT
通讯作者：
Washington MT
影响因子：
64.8
作者：
Cuylen S;Blaukopf C;Politi AZ;Müller-Reichert T;Neumann B;Poser I;Ellenberg J;Hyman AA;Gerlich DW
通讯作者：
Gerlich DW
影响因子：
2.1
作者：
Dick, CAJ;Brown, DM;Stone, V
通讯作者：
Stone, V
影响因子：
3.9
作者：
Alarifi, Saud;Ali, Daoud;Al-Qahtani, Ahmed A.
通讯作者：
Al-Qahtani, Ahmed A.