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端粒酶 siRNA 诱导神经母细胞瘤细胞核浓缩和细胞周期停滞。

Nuclear condensation and cell cycle arrest induced by telomerase siRNA in neuroblastoma cells.

机构信息

Graduate Institute of Clinical Medicine, College of Medicine, National Taiwan University, Taipei, Taiwan.

出版信息

J Neurooncol. 2013 Feb;111(3):265-72. doi: 10.1007/s11060-012-1025-y. Epub 2012 Dec 14.

Abstract

Neuroblastoma is a type of malignant extracranial tumor that occurs in children. Advanced neuroblastoma, and tumors with MYCN amplification in particular, have poor prognoses. Therefore, it is important to find an effective cure for this disease. Small interfering RNA (siRNA) disrupts gene function by specifically binding to target mRNA. In this study, we used siRNA against telomerase to treat neuroblastoma, to evaluate any anti-proliferative effect on these cells. We evaluated cell viability by WST-1 assay on neuroblastoma cells treated with or without telomerase siRNA. Nuclear condensation, an indicator for apoptotic cells, was determined by DAPI labeling following siRNA treatment. The effectiveness of telomerase siRNA on altering the neuroblastoma cell cycle was detected by flow cytometry. Our results indicated that telomerase siRNA reduces the viability of neuroblastoma cells and increases the percentage of cells in the cell cycle's sub-G1 phase. We found that telomerase siRNA increases the percentage of condensed DNA in neuroblastoma cells. In conclusion, using siRNA against telomerase could be further developed as a therapy for the treatment of neuroblastoma.

摘要

神经母细胞瘤是一种发生在儿童身上的恶性颅外肿瘤。晚期神经母细胞瘤,特别是具有 MYCN 扩增的肿瘤,预后较差。因此,找到治疗这种疾病的有效方法非常重要。小干扰 RNA(siRNA)通过特异性结合靶 mRNA 来破坏基因功能。在这项研究中,我们使用针对端粒酶的 siRNA 治疗神经母细胞瘤,以评估其对这些细胞的任何抗增殖作用。我们通过 WST-1 assay 评估了用或不用端粒酶 siRNA 处理的神经母细胞瘤细胞的活力。用 DAPI 标记法在 siRNA 处理后测定核浓缩,这是凋亡细胞的一个指标。通过流式细胞术检测端粒酶 siRNA 对改变神经母细胞瘤细胞周期的效果。我们的结果表明,端粒酶 siRNA 降低了神经母细胞瘤细胞的活力,并增加了细胞周期的 sub-G1 期细胞的比例。我们发现端粒酶 siRNA 增加了神经母细胞瘤细胞中浓缩 DNA 的比例。总之,使用针对端粒酶的 siRNA 可能会进一步发展为治疗神经母细胞瘤的一种疗法。

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