Telomerase inhibition, telomere shortening, cell growth suppression and induction of apoptosis by telomestatin in childhood neuroblastoma cells
Telomerase inhibition, telomere shortening, cell growth suppression and induction of apoptosis by telomestatin in childhood neuroblastoma cells
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DOI：
10.1016/j.ejca.2005.08.025
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发表时间：
2005-12-01
影响因子：
8.4
通讯作者：
Grotzer, MA
中科院分区：
文献类型：
--
作者：
Binz, N;Shalaby, T;Grotzer, MA
神经母细胞瘤是一种源自交感神经系统原始细胞的肿瘤，是儿童期最常见的颅外实体瘤。不利肿瘤不仅表现为结构变化，包括MYCN原癌基因的1p缺失和扩增，还表现为端粒酶活性高。富含端粒g的单链DNA在体外可以采用分子内四重结构，这已被证明可以抑制端粒酶的活性。在这项研究中，我们检测了端粒抑素（一种g -四重体相互作用剂）抑制神经母细胞瘤细胞端粒维持的能力。端粒长度测定采用末端限制性内切片段法，端粒酶活性测定采用定量端粒重复扩增法，人端粒酶表达采用实时定量聚合酶链反应（RTPCR）法。短期使用端米司汀治疗可导致剂量依赖性细胞毒性和诱导细胞凋亡。长期使用端粒抑素进行无细胞毒性治疗，但仍然抑制端粒酶活性，浓度导致端粒缩短、生长停滞和诱导细胞凋亡。这些结果表明，端米抑素的作用是剂量依赖性的，至少是2倍的。长时间的低剂量端米司汀治疗通过破坏端粒维持限制NB细胞的细胞寿命。(c) 2005 Elsevier Ltd版权所有。
Neuroblastoma is a tumour derived from primitive cells of the sympathetic nervous system and is the most common extracranial solid tumour in childhood. Unfavourable tumours are characterised not only by structural changes, including 1 p deletion and amplification of the MYCN proto-oncogene, but also by high telomerase activity. Telomeric G-rich single-stranded DNA can adopt in vitro an intramolecular quadruplex structure, which has been shown to inhibit telomerase activity. In this study, we examined telomestatin, a G-quadruplex interactive agent, for its ability to inhibit telomere maintenance of neuroblastoma cells. Telomere length was determined by the terminal restriction fragment method, telomerase activity was measured by a quantitative telomeric repeat amplification protocol, and the expression of human telomerase by quantitative real-time polymerase chain reaction (RTPCR). Short-term treatment with telomestatin resulted in dose-dependent cytotoxicity and induction of apoptosis. Long-term treatment with telomestatin at non-cytotoxic, but still telomerase activity-inhibiting, concentrations resulted in telomere shortening, growth arrest and induction of apoptosis. These results suggest that the effect of telomestatin is dose-dependent and at least 2-fold. Prolonged low-dose treatment with telomestatin limits the cellular lifespan of NB cells through disruption of telomere maintenance. (c) 2005 Elsevier Ltd. All rights reserved.