核干细胞因子缺失诱导慢性粒细胞白血病K562细胞G1期后停滞凋亡。
Nucleostemin depletion induces post-g1 arrest apoptosis in chronic myelogenous leukemia k562 cells.
作者信息
Seyed-Gogani Negin, Rahmati Marveh, Zarghami Nosratollah, Asvadi-Kermani Iraj, Hoseinpour-Feyzi Mohammad Ali, Moosavi Mohammad Amin
机构信息
Department of Zoology, Faculty of Natural Science, University of Tabriz, Tabriz, Iran.
Department of Biochemistry, Faculty of Medicine, Tabriz University of Medical Science, Tabriz, Iran.
出版信息
Adv Pharm Bull. 2014;4(1):55-60. doi: 10.5681/apb.2014.009. Epub 2013 Dec 23.
PURPOSE
Despite significant improvements in treatment of chronic myelogenous leukemia (CML), the emergence of leukemic stem cell (LSC) concept questioned efficacy of current therapeutical protocols. Remaining issue on CML includes finding and targeting of the key genes responsible for self-renewal and proliferation of LSCs. Nucleostemin (NS) is a new protein localized in the nucleolus of most stem cells and tumor cells which regulates their self-renewal and cell cycle progression. The aim of this study was to investigate effects of NS knocking down in K562 cell line as an in vitro model of CML.
METHODS
NS gene silencing was performed using a specific small interfering RNA (NS-siRNA). The gene expression level of NS was evaluated by RT-PCR. The viability and growth rate of K562 cells were determined by trypan blue exclusion test. Cell cycle distribution of the cells was analyzed by flow cytometry.
RESULTS
Our results showed that NS knocking down inhibited proliferation and viability of K562 cells in a time-dependent manner. Cell cycle studies revealed that NS depletion resulted in G(1) cell cycle arrest at short times of transfection (24 h) followed with apoptosis at longer times (48 and 72 h), suggest that post-G1 arrest apoptosis is occurred in K562 cells.
CONCLUSION
Overall, these results point to essential role of NS in K562 cells, thus, this gene might be considered as a promising target for treatment of CML.
目的
尽管慢性粒细胞白血病（CML）的治疗有了显著改善，但白血病干细胞（LSC）概念的出现对当前治疗方案的疗效提出了质疑。CML 尚存的问题包括寻找并靶向负责 LSC 自我更新和增殖的关键基因。核仁素（NS）是一种定位于大多数干细胞和肿瘤细胞核仁的新蛋白，它调节细胞的自我更新和细胞周期进程。本研究旨在探讨在作为 CML 体外模型的 K562 细胞系中敲低 NS 的影响。
方法
使用特异性小干扰 RNA（NS-siRNA）进行 NS 基因沉默。通过 RT-PCR 评估 NS 的基因表达水平。通过台盼蓝排斥试验测定 K562 细胞的活力和生长速率。通过流式细胞术分析细胞的细胞周期分布。
结果
我们的结果表明，敲低 NS 以时间依赖性方式抑制 K562 细胞的增殖和活力。细胞周期研究显示，NS 缺失在转染后短时间（24 小时）导致 G(1)期细胞周期停滞，随后在较长时间（48 和 72 小时）发生凋亡，这表明 K562 细胞中发生了 G1 期后停滞诱导的凋亡。
结论
总体而言，这些结果表明 NS 在 K562 细胞中起重要作用，因此，该基因可能被视为治疗 CML 的一个有前景的靶点。
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