Role of survivin in acute lung injury: epithelial cells of mice and humans
Role of survivin in acute lung injury: epithelial cells of mice and humans
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DOI：
10.1038/labinvest.2013.103
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发表时间：
2013-10-01
影响因子：
5
通讯作者：
Fukuda, Yuh
中科院分区：
文献类型：
--
作者：
Terasaki, Yasuhiro;Terasaki, Mika;Fukuda, Yuh
存活素是一种凋亡抑制剂，调节细胞分裂，是抗癌药物的潜在靶点，因为许多癌症表达高水平的存活素。然而，存活素是否会对人类肺细胞和组织有毒尚未确定。本报告阐明了生存素在急性肺损伤中的作用。我们使用免疫组化分析，免疫电镜，实时逆转录-定量聚合酶链反应，研究存活素的表达和定位在受伤的小鼠和人的肺。我们还使用培养的人肺上皮细胞（BEAS-2B和A549）来研究生存素的细胞保护作用。博来霉素损伤肺模型第3天和第7天的上皮细胞的细胞核和细胞质显示Survivin阳性结果，这与Survivin nnRNA表达上调一致。这些细胞核也证明了增殖细胞核抗原和生存素的双阳性结果。第7天模型具有相似的Smac/DIABLO阳性和存活素阳性细胞分布。在弥漫性肺泡损伤的病变中，上皮细胞的细胞质和细胞核表现出强阳性结果。博来霉素刺激两种上皮细胞系上调生存素和凋亡相关分子的表达。与模拟小干扰RNA处理的细胞相比，用小干扰RNA抑制生存素表达使人肺上皮细胞对博来霉素诱导的损伤敏感，caspase-3、caspase-7、聚（ADP-核糖）聚合酶和乳酸脱氢酶活性的活化显著上调，死亡细胞的数量增加。通过转染过表达生存素导致这些上皮细胞对博来霉素诱导的细胞损伤具有抗性，与模拟转染细胞的结果相比，凋亡相关分子和乳酸脱氢酶活性的活化减少，死亡细胞减少。存活素在上皮细胞水平上起作用，部分依赖于凋亡抑制，因此是急性肺损伤中细胞保护的关键介质。了解生存素在正常肺细胞中的确切作用是开发治疗性生存素所必需的。
Survivin, an inhibitor of apoptosis, regulates cell division and is a potential target for anticancer drugs because many cancers express high survivin levels. However, whether survivin would be toxic to human lung cells and tissues has not been determined. This report clarified the involvement of survivin in acute lung injury. We used immunohistochemical analysis, immunoelectron microscopy, and real-time reverse transcription-quantitative polymerase chain reaction to study survivin expression and localization in injured mouse and human lungs. We also used cultured human lung epithelial cells (BEAS-2B and A549) to study survivin cytoprotection. Nuclei and cytoplasm of epithelial cells in day 3 and day 7 models of bleomycin-injured lung showed survivin-positive results, which is consistent with upregulated survivin nnRNA expression. These nuclei also evidenced double positive findings for proliferating cell nuclear antigen and survivin. Day 7 models had similar Smac/DIABLO-positive and survivin-positive cell distributions. The cytoplasm and nuclei of epithelial cells in lesions with diffuse alveolar damage manifested strong survivin-positive findings. Bleomycin stimulation in both epithelial cell lines upregulated expression of survivin and apoptosis-related molecules. Suppression of survivin expression with small interfering RNA rendered human lung epithelial cells susceptible to bleomycin-induced damage, with markedly upregulated activation of caspase-3, caspase-7, poly (ADP-ribose) polymerase, and lactate dehydrogenase activity and an increased number of dead cells compared with mock small interfering RNA-treated cells. Overexpression of survivin via transfection resulted in these epithelial cells being resistant to bleomycin-induced cell damage, with reduced activation of apoptosis-related molecules and lactate dehydrogenase activity and fewer dead cells compared with results for mock-transfected cells. Survivin, acting at the epithelial cell level that depends partly on apoptosis inhibition, is therefore a key mediator of cytoprotection in acute lung injury. Understanding the precise role of survivin in normal lung cells is required for the development of therapeutic survivin.