The ovarian DNA damage repair response is induced prior to phosphoramide mustard-induced follicle depletion, and ataxia telangiectasia mutated inhibition prevents PM-induced follicle depletion
The ovarian DNA damage repair response is induced prior to phosphoramide mustard-induced follicle depletion, and ataxia telangiectasia mutated inhibition prevents PM-induced follicle depletion
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DOI：
10.1016/j.taap.2015.12.010
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发表时间：
2016-02-01
影响因子：
3.8
通讯作者：
Keating, Aileen F.
中科院分区：
文献类型：
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作者：
Ganesan, Shanthi;Keating, Aileen F.
磷酰胺芥（PM）是环磷酰胺的一种卵毒性代谢产物，可通过诱导DNA损伤破坏原始卵泡和初级卵泡。PM诱导DNA损伤和启动卵巢对DNA损伤的反应的时间模式尚未得到很好的表征。本研究使用新生大鼠卵巢培养系统研究了DNA损伤的启动、DNA修复反应以及卵泡死亡的诱导。此外，为了描述卵巢对PM暴露的反应中涉及的特定机制，使用PKC δ（PKC δ）缺陷小鼠以及ATM抑制剂（KU 55933; AI）。将Fisher 344 PND 4大鼠卵巢在含有DMSO +/- 60 μ M PM或KU 55933（48 h; 10 nM）的培养基中培养12、24、48或96 h。早在暴露后12 h就观察到PM诱导的DNA损伤修复基因的激活。PM暴露96 h后，ATM、PARP 1、E2 F7、P73和CASP 3蛋白表达量增加，而RAD 51和BCL 2蛋白表达量减少。PKC δ缺乏减少了所有卵泡阶段的数量，但对PM诱导的卵毒性没有附加影响。ATM抑制保护所有卵泡阶段从PM诱导的消耗。总之，卵巢DNA损伤修复反应是积极的PM暴露后，支持DNA损伤有助于PM诱导的卵毒性。(C)2015爱思唯尔公司All rights reserved.
Phosphoramide mustard (PM) is an ovotoxic metabolite of cyclophosphamide and destroys primordial and primary follicles potentially by DNA damage induction. The temporal pattern by which PM induces DNA damage and initiation of the ovarian response to DNA damage has not yet been well characterized. This study investigated DNA damage initiation, the DNA repair response, as well as induction of follicular demise using a neonatal rat ovarian culture system. Additionally, to delineate specific mechanisms involved in the ovarian response to PM exposure, utility was made of PKC delta (PKC delta) deficient mice as well as an ATM inhibitor (KU 55933; AI). Fisher 344 PND4 rat ovaries were cultured for 12, 24, 48 or 96 h in medium containing DMSO +/- 60 mu M PM or KU 55933 (48 h; 10 nM). PM-induced activation of DNA damage repair genes was observed as early as 12 h post exposure. ATM, PARP1, E2F7, P73 and CASP3 abundance were increased but RAD51 and BCL2 protein decreased after 96 h of PM exposure. PKC delta deficiency reduced numbers of all follicular stages, but did not have an additive impact on PM-induced ovotoxicity. ATM inhibition protected all follicle stages from PM-induced depletion. In conclusion, the ovarian DNA damage repair response is active post-PM exposure, supporting that DNA damage contributes to PM-induced ovotoxicity. (C) 2015 Elsevier Inc. All rights reserved.