Han Wei, Zhu Lingyan, Jiang Erkang, Wang Jun, Chen Shaopeng, Bao Linzhi, Zhao Ye, Xu An, Yu Zengliang, Wu Lijun
Key Laboratory of Ion Beam Bioengineering, Institute of Plasma Physics, Chinese Academy of Sciences, Hefei 230031, People's Republic of China.
Mutat Res. 2007 Nov 1;624(1-2):124-31. doi: 10.1016/j.mrfmmm.2007.04.010. Epub 2007 May 5.
Previous studies have shown that high NaCl can be genotoxic, either alone or combined with irradiation. However, little is known about the relationship between environmental NaCl at elevated conditions and radiation-induced bystander effects (RIBE). RIBE, which has been considered as non-targeted bystander responses, has been demonstrated to occur widely in various cell lines. In the present study, RIBE under the elevated NaCl culture condition was assessed in AG 1522 cells by both the induction of gamma-H2AX, a reliable marker of DNA double-strand break (DSB) for the early process (<1h post irradiation), and the generation of micronuclei (MN), a sensitive marker for relative long process of RIBE. Our results showed that in the absence of irradiation, NaCl at elevated concentration such as 8.0, 9.0 and 10.0g/L did not significantly increase the frequency of gamma-H2AX foci-positive cells and the number of foci per positive cell comparing with that NaCl at a normal concentration (6.8g/L). However, with 0.2cGy alpha-particle irradiation, the induced fraction of gamma-H2AX foci-positive cells and the number of induced gamma-H2AX foci per positive cell were significantly increased in both irradiated and adjacent non-irradiated regions. Similarly, the induction of MN by 0.2cGy alpha-particle irradiation also increased with the elevated NaCl concentrations. With N(G)-methyl-l-arginine, an inhibitor of nitric oxide synthase, the induced fraction of foci-positive cells was effectively inhibited both in 0.2cGy alpha-particle irradiated and adjacent non-irradiated regions under either normal or elevated NaCl conditions. These results suggested that the cultures with elevated NaCl medium magnified the damage effects induced by the low dose alpha-particle irradiation and nitric oxide generated by irradiation was also very important in this process.
先前的研究表明,高浓度氯化钠无论是单独存在还是与辐射联合,都可能具有基因毒性。然而,关于环境中高浓度氯化钠与辐射诱导的旁观者效应(RIBE)之间的关系,人们所知甚少。RIBE被认为是一种非靶向性的旁观者反应,已被证明在各种细胞系中广泛存在。在本研究中,通过检测γ-H2AX(DNA双链断裂(DSB)早期过程(照射后<1小时)的可靠标志物)的诱导情况以及微核(MN)的产生(RIBE相对较长过程的敏感标志物),评估了AG 1522细胞在高浓度氯化钠培养条件下的RIBE。我们的结果显示,在无辐射情况下,与正常浓度(6.8g/L)的氯化钠相比,8.0、9.0和10.0g/L等较高浓度的氯化钠并未显著增加γ-H2AX焦点阳性细胞的频率以及每个阳性细胞的焦点数量。然而,在0.2cGyα粒子照射后,照射区域和相邻未照射区域中γ-H2AX焦点阳性细胞的诱导比例以及每个阳性细胞诱导的γ-H2AX焦点数量均显著增加。同样,0.2cGyα粒子照射诱导的微核也随着氯化钠浓度的升高而增加。使用一氧化氮合酶抑制剂N(G)-甲基-L-精氨酸后,在正常或高浓度氯化钠条件下,0.2cGyα粒子照射区域和相邻未照射区域中焦点阳性细胞的诱导比例均受到有效抑制。这些结果表明,高浓度氯化钠培养基培养放大了低剂量α粒子照射诱导的损伤效应,并且照射产生的一氧化氮在此过程中也非常重要。