Biological and molecular mechanisms of sulfur mustard analogue-induced toxicity in JB6 and HaCaT cells: possible role of ataxia telangiectasia-mutated/ataxia telangiectasia-Rad3-related cell cycle checkpoint pathway.
Biological and molecular mechanisms of sulfur mustard analogue-induced toxicity in JB6 and HaCaT cells: possible role of ataxia telangiectasia-mutated/ataxia telangiectasia-Rad3-related cell cycle checkpoint pathway.
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DOI：
10.1021/tx100038b
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发表时间：
2010-06-21
影响因子：
4.1
通讯作者：
Agarwal, Rajesh
中科院分区：
文献类型：
--
作者：
Tewari-Singh, Neera;Gu, Mallikarjuna;Agarwal, Chapla;White, Carl W.;Agarwal, Rajesh
由于对化学战剂硫芥子气的作用机制了解有限，因此缺乏有效的医疗和预防措施来治疗其引起的失能性皮肤毒性。在HD引起的皮肤损伤中，增殖的基底表皮细胞是主要的攻击部位。因此，采用小鼠JB 6和人HaCaT表皮细胞，在这里，我们研究了HD类似物，2-氯乙基乙基硫醚（CEES）诱导的皮肤细胞毒性的分子机制。与对照相比，高达1 mM CEES处理这些细胞2、4和24 h，导致细胞活力和增殖的剂量依赖性降低，如通过DNA合成所测量的，以及细胞周期进程中的S和G2-M期阻滞。机制研究表明，磷酸化的DNA损伤传感器和检查点激酶，ATM在ser 1981和ATR在ser 428在30分钟内CEES曝光，和S和G2-M期相关的细胞周期调控蛋白的ATM和ATR激酶的下游目标的调制。Hoechst-碘化丙啶（PI）染色表明，CEES诱导的细胞死亡是坏死和凋亡的性质，后者是在4和24小时的CEES处理HaCaT和JB 6细胞，分别诱导。caspase-3活性的增加以及caspase-3和PARP裂解与CEES引起的细胞凋亡一致，表明caspase途径参与。总之，我们的研究结果表明，CEES的DNA损伤作用，激活ATM/ATR-细胞周期检查点信号传导以及半胱天冬酶-PARP通路，导致JB 6和HaCaT细胞的细胞周期停滞和凋亡/坏死。本研究中确定的分子靶点、定量生物标志物和表皮细胞模型在快速开发针对HD诱导的皮肤毒性的有效预防和治疗干预方面具有潜力和实用性。
Effective medical treatment and preventive measures for chemical warfare agent sulfur mustard (HD)-caused incapacitating skin toxicity are lacking, owing to limited knowledge of its mechanism of action. The proliferating basal epidermal cells are primary major sites of attack during HD-caused skin injury. Therefore, employing mouse JB6 and human HaCaT epidermal cells, here we investigated the molecular mechanism of HD analog, 2-chloroethyl ethyl sulfide (CEES)-induced skin cytotoxicity. As compared to control, up to 1 mM CEES treatment of these cells for 2, 4 and 24 h, caused dose-dependent decreases in cell viability and proliferation as measured by DNA synthesis, together with S and G2-M phase arrest in cell cycle progression. Mechanistic studies showed phosphorylation of DNA damage sensors and checkpoint kinases, ATM at ser1981 and ATR at ser428 within 30 min of CEES exposure, and modulation of S and G2-M phase-associated cell cycle regulatory proteins which are downstream targets of ATM and ATR kinases. Hoechst-propidium iodide (PI) staining demonstrated that CEES-induced cell death was both necrotic and apoptotic in nature and latter was induced at 4 and 24 h of CEES treatment in HaCaT and JB6 cells, respectively. An increase in caspase-3 activity and both caspase-3 and PARP cleavage coinciding with CEES-caused apoptosis in both cell lines suggested the involvement of caspase pathway. Together, our findings suggest a DNA damaging effect of CEES that activates ATM/ATR-cell cycle checkpoint signaling as well as caspase-PARP pathways leading to cell cycle arrest and apoptosis/necrosis in both JB6 and HaCaT cells. The identified molecular targets, quantitative biomarkers and epidermal cell models in this study, have the potential and usefulness in rapid development of effective prophylactic and therapeutic interventions against HD-induced skin toxicity.
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DOI：
10.1124/jpet.108.145037
发表时间：
2009-03-01
影响因子：
3.5
作者：
Gould, Neal S.;White, Carl W.;Day, Brian J.
通讯作者：
Day, Brian J.
影响因子：
3.8
作者：
Brodsky, Berta;Erlanger-Rosengarten, Avigail;Wormser, Uri
通讯作者：
Wormser, Uri
影响因子：
3.7
作者：
Kurose, A;Tanaka, T;Darzynkiewicz, Z
通讯作者：
Darzynkiewicz, Z
影响因子：
4.5
作者：
Kehe, K;Szinicz, L
通讯作者：
Szinicz, L
影响因子：
5.7
作者：
Kiziltepe, Tanyel;Hideshima, Teru;Anderson, Kenneth C.
通讯作者：
Anderson, Kenneth C.