Thapsigargin induces apoptosis in SH‐SY5Y neuroblastoma cells and cerebrocortical cultures
Thapsigargin induces apoptosis in SH‐SY5Y neuroblastoma cells and cerebrocortical cultures
复制标题
毒胡萝卜素诱导 SH-SY5Y 神经母细胞瘤细胞和脑皮质培养物凋亡
DOI：
10.1080/15216549700203971
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发表时间：
1997
期刊：
影响因子：
4.6
通讯作者：
Kevin K. W. Wang
中科院分区：
文献类型：
--
作者：
R. Nath;K. Raser;I. Hajimohammadreza;Kevin K. W. Wang
Thapsigargin是一种内质网状Ca2+‐atp酶的特异性抑制剂，以前曾被用于动员细胞内钙储存的钙释放。我们现在发现thapsigargin（1‐10 μM）在神经母细胞瘤细胞系（SH‐SY5Y）和胎鼠脑皮质培养物中诱导细胞凋亡。用thapsigargin治疗24 - 48小时后，用乳酸脱氢酶释放法观察细胞死亡。在这两种情况下，从thapsigargin处理的细胞中提取的DNA显示阶梯状，这是典型的核内切酶介导的核小体间分裂。DNA片段的存在也被设计用于检测凋亡细胞核小体的ELISA证实。环己亚胺降低了thapsigargin处理细胞的DNA断裂和损伤程度。Dantrolene，一种钙从细胞内储存释放的抑制剂，部分消除了thapsigargin的作用，这表明最初的Ca2+升高可能是凋亡细胞死亡途径中的信号事件。我们认为，在培养的神经细胞中，thapsigargin诱导的细胞死亡可能是研究凋亡的分子和遗传事件的一个有用的系统。
Thapsigargin, a specific inhibitor of the endoplasmic reticular Ca2+‐ATPase, has been used previously to mobilize calcium release from intracellular calcium stores. We now show that thapsigargin (1‐10 μM) induces apoptosis in a neuroblastoma cell line (SH‐SY5Y) and in fetal rat cerebrocortical cultures. Cell death measured by lactate dehydrogenase release was observed 24‐48 hours after treatment with thapsigargin. In both cases, DNA extracts from thapsigargin treated cells showed laddering, typical of endonuclease‐mediated internucleosomal cleavages. The presence of DNA fragments was also confirmed by an ELISA designed for detecting nucleosomes in apoptotic cells. Cycloheximide reduced the extent of DNA fragmentation and injury in thapsigargin‐treated cells. Dantrolene, an inhibitor of calcium release from intracellular stores partially abolished the effect of thapsigargin, suggesting that the initial Ca2+ rise may be the signalling event in this apoptotic cell death pathway. We propose that thapsigargin‐induced cell death in cultured neuronal cells maybe a useful system to study the molecular and genetic events involved in apoptosis.