Alcohol exposure alters cell cycle and apoptotic events during early neurulation
Alcohol exposure alters cell cycle and apoptotic events during early neurulation
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DOI：
10.1093/alcalc/agm166
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发表时间：
2008-05-01
影响因子：
2.8
通讯作者：
Ruiz, Joseph
中科院分区：
文献类型：
--
作者：
Anthony, Bruce;Zhou, Feng C.;Ruiz, Joseph
背景：胎儿酒精暴露可导致生长缺陷、小脑畸形和神经系统异常。虽然酒精对发育延迟和生长相关缺陷的影响已经被假设，但很少有人了解酒精如何改变，特别是细胞周期内的细胞周期蛋白通路，这对增殖和凋亡控制至关重要。在这项研究中，我们研究了细胞周期蛋白相关的G1-S期转换和凋亡，以确定细胞周期失调是否可以归因于凋亡诱导和生长缺陷。研究方法：我们研究了细胞周期调控在G1期和S期，和DNA断裂损伤，使用E14背根神经节神经干细胞（DRG-NC），并培养小鼠胚胎暴露于200和400毫克/分升乙醇。结果如下：暴露于酒精的DRG-NC表现出细胞周期蛋白D1蛋白表达增加和DNA片段化增加的剂量依赖性增加。使用胚胎进行的Western印迹分析表明，细胞周期蛋白D1、D2和E2 F1（G1至S期细胞周期调控的关键成分）过度表达，p53增加，连接细胞周期和凋亡途径。溴脱氧尿苷掺入表明在几个胚胎区域的DNA合成和生长减少。碘化丙啶染色显示，在几个胚胎组织中，DNA含量减少，DNA片段增加。结论：这项研究表明，延迟生长的DRG-NC和胚胎，由酒精诱导，与细胞周期和凋亡蛋白的表达改变，并同时抑制增殖和增加DNA片段。我们认为，酒精诱导细胞周期蛋白D1的表达增加，过早的S期进入，DNA合成脱节与细胞凋亡增加。
Background: Fetal alcohol exposure causes growth deficits, microencephaly, and neurological abnormalities. Although the effects of alcohol on developmental delay and growth-related deficits have been hypothesized, little is understood about how alcohol alters, in particular, the cyclin pathway within the cell cycle, which is critical to proliferation and apoptotic control. In this study, we examined cell cycle proteins pertinent to the G1-S phase transition and apoptosis, to determine if cell cycle misregulation can be attributed to apoptotic induction and growth defects. Methods: We examined cell cycle regulation during G1 and S-phase, and DNA fragmentation damage, using E14 dorsal root ganglia neural stem cells (DRG-NC), and cultured mouse embryos exposed to 200 and 400 mg/dl ethanol. Results: Alcohol-exposed DRG-NC demonstrated a dose-dependent increase in cells expressing increased cyclin D1 protein, and increased DNA fragmentation. Western blot analysis, using embryos, demonstrated an overexpression of cyclin D1, D2, and E2F1, key G1 to S-phase cell cycle regulatory components, and increases in p53, linking the cell cycle and apoptotic pathways. Bromodeoxyuridine incorporation indicated reduced DNA synthesis and growth in several embryonic regions. Propidium iodide staining demonstrated decreases in DNA content and increases in DNA fragmentation in several embryonic tissues. Conclusions: This study indicated that retarded growth of DRG-NC and embryos, induced by alcohol, is associated with altered expression of cell cycle and apoptotic proteins and concurrent inhibition of proliferation and increased DNA fragmentation. We suggest that alcohol induces an increase in cyclin D1 expression, premature S-phase entry, and disjointed DNA synthesis with increased apoptosis.