Activation of DNA damage repair pathways in response to nitrogen mustard-induced DNA damage and toxicity in skin keratinocytes.
Activation of DNA damage repair pathways in response to nitrogen mustard-induced DNA damage and toxicity in skin keratinocytes.
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DOI：
10.1016/j.mrfmmm.2014.04.002
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发表时间：
2014-05
影响因子：
2.3
通讯作者：
Agarwal, Rajesh
中科院分区：
文献类型：
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作者：
Inturi, Swetha;Tewari-Singh, Neera;Agarwal, Chapla;White, Carl W.;Agarwal, Rajesh
氮芥（NM）是化学战剂硫芥（SM）的结构类似物，可与DNA、RNA和蛋白质形成加合物和交联物。在这里，我们研究了NM诱导的皮肤毒性的机制，响应于双链断裂（DSB），导致细胞周期停滞，以促进DNA修复，作为一个模型，开发对抗水疱剂诱导的皮肤损伤的对策。NM暴露的小鼠表皮JB 6细胞的细胞生长下降，并导致S期阻滞。与这些生物学结果一致，NM暴露也增加了彗星尾延伸矩和DNA DSB修复分子磷酸化H2A.X Ser 139和p53 Ser 15的水平，表明NM诱导的DNA DSB。由于DNA DSB修复通过非同源末端连接途径（NHEJ）或同源重组修复（HRR）途径发生，接下来我们研究了这两种途径，并注意到它们的激活，分别通过磷酸化和总DNA-PK水平的增加以及Rad 51灶的形成来定义。为了进一步分析这些途径在细胞对NM诱导的细胞毒性反应中的作用，NHEJ和HRR分别被DNA-PK抑制剂NU 7026和Rad 51抑制剂BO 2抑制。NHEJ的抑制并不使细胞对NM诱导的细胞生长和细胞周期停滞的减少敏感。然而，HRR通路的抑制引起细胞死亡的显着增加，并延长NM暴露后的G2 M期阻滞。总之，我们的研究结果表明，HRR是参与NM诱导的DNA DSB修复的关键途径，可能有助于开发新的治疗策略来对抗水疱剂诱导的皮肤损伤。
Nitrogen mustard (NM), a structural analog of chemical warfare agent sulfur mustard (SM), forms adducts and crosslinks with DNA, RNA and proteins. Here we studied the mechanism of NM-induced skin toxicity in response to double strand breaks (DSBs) resulting in cell cycle arrest to facilitate DNA repair, as a model for developing countermeasures against vesicant-induced skin injuries. NM exposure of mouse epidermal JB6 cells decreased cell growth and caused S-phase arrest. Consistent with these biological outcomes, NM exposure also increased comet tail extent moment and the levels of DNA DSB repair molecules phospho H2A.X Ser139 and p53 Ser15 indicating NM-induced DNA DSBs. Since DNA DSB repair occurs via non homologous end joining pathway (NHEJ) or homologous recombination repair (HRR) pathways, next we studied these two pathways and noted their activation as defined by an increase in phospho- and total DNA-PK levels, and the formation of Rad51 foci, respectively. To further analyze the role of these pathways in the cellular response to NM-induced cytotoxicity, NHEJ and HRR were inhibited by DNA-PK inhibitor NU7026 and Rad51 inhibitor BO2, respectively. Inhibition of NHEJ did not sensitize cells to NM-induced decrease in cell growth and cell cycle arrest. However, inhibition of the HRR pathway caused a significant increase in cell death, and prolonged G2M arrest following NM exposure. Together, our findings, indicating that HRR is the key pathway involved in the repair of NM-induced DNA DSBs, could be useful in developing new therapeutic strategies against vesicant-induced skin injury.
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影响因子：
5.3
作者：
Ding, Q;Reddy, YVR;Meek, K
通讯作者：
Meek, K
DOI：
10.1073/pnas.1003449107
发表时间：
2010-05-04
影响因子：
11.1
作者：
Callegari, A. John;Clark, Emily;Kelly, Thomas J.
通讯作者：
Kelly, Thomas J.
影响因子：
3.6
作者：
GOLUMBIC, C;FRUTON, JS;BERGMANN, M
通讯作者：
BERGMANN, M
影响因子：
3.8
作者：
KU, WW;BERNSTEIN, IA
通讯作者：
BERNSTEIN, IA
DNA-PKcs 在响应 DNA 损伤和细胞周期进展的 H2AX 磷酸化调节中起主导作用。
影响因子：
--
作者：
An J;Huang YC;Xu QZ;Zhou LJ;Shang ZF;Huang B;Wang Y;Liu XD;Wu DC;Zhou PK
通讯作者：
Zhou PK