Controlling subcellular localization to alter function: Sending oncogenic Bcr-Abl to the nucleus causes apoptosis.
Controlling subcellular localization to alter function: Sending oncogenic Bcr-Abl to the nucleus causes apoptosis.
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DOI：
10.1016/j.jconrel.2009.06.026
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发表时间：
2009-12-16
影响因子：
10.8
通讯作者：
Lim, Carol S.
中科院分区：
文献类型：
--
作者：
Dixon, Andrew S.;Kakar, Mudit;Schneider, Korbinian M. H.;Constance, Jonathan E.;Paullin, Blake C.;Lim, Carol S.
改变信号转导蛋白的亚细胞定位是一种新的治疗干预方法。肿瘤抑制因子、癌基因或参与凋亡的因子的错误定位导致这些蛋白质的异常功能，从而导致疾病。在慢性粒细胞白血病（CML）的情况下，细胞质Bcr-Abl引起肿瘤发生/增殖。另一方面，内源性Bcr-Abl（在K562人白血病细胞中）的核截留引起细胞凋亡。本研究的目的是确定表达Bcr-Abl的质粒是否可以通过强核定位信号（NLS）特异性地定向到细胞核时引起K562细胞的凋亡。将来自SV 40大T抗原的单个NLS或四个NLS亚克隆至Bcr-Abl（1 NLS-Bcr-Abl或4 NLS-Bcr-Abl）。转染K562细胞后，只有4 NLS Bcr Abl易位到细胞核。Bcr-Abl单独被发现定位于细胞质中，由于其肌动蛋白结合结构域而与肌动蛋白共定位。1 NLS-Bcr-Abl也定位于肌动蛋白。转染后24 h，通过形态学观察、DNA染色和caspase-3检测，评价4 NLS-Bcr-Abl诱导的细胞凋亡。这是首次证明改变表达Bcr-Abl的质粒的位置可以杀死白血病细胞。需要多个NLS来克服Bcr-Abl与肌动蛋白的结合，从而将其驱动到细胞核中并引起细胞凋亡。
Altering the subcellular localization of signal transducing proteins is a novel approach for therapeutic intervention. Mislocalization of tumor suppressors, oncogenes, or factors involved in apoptosis result in aberrant functioning of these proteins, leading to disease. In the case of chronic myelogenous leukemia (CML), cytoplasmic Bcr-Abl causes oncogenesis/proliferation. On the other hand, nuclear entrapment of endogenous Bcr-Abl (in K562 human leukemia cells) causes apoptosis. The goal of this study was to determine whether plasmid expressed Bcr-Abl could cause apoptosis of K562 cells when specifically directed to the nucleus via strong nuclear localization signals (NLSs). A single NLS from SV40 large T-antigen or four NLSs were subcloned to Bcr-Abl (1NLS-Bcr-Abl or 4NLS-Bcr-Abl). When transfected into K562 cells, only 4NLS-Bcr-Abl translocated to the nucleus. Bcr-Abl alone was found to localize in the cell cytoplasm, colocalizing with actin due to its actin binding domain. 1NLS-Bcr-Abl also localized with actin. Apoptosis induced by 4NLS-Bcr-Abl was evaluated 24 hours post-transfection by morphologic determination, DNA staining, and caspase-3 assay. This is the first demonstration that altering the location of plasmid expressed Bcr-Abl can kill leukemia cells. Multiple NLSs are required to overcome Bcr-Abl binding to actin, thus driving it into the nucleus and causing apoptosis.
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