8-Chloro-adenosine inhibits growth at least partly by interfering with actin polymerization in cultured human lung cancer cells
8-Chloro-adenosine inhibits growth at least partly by interfering with actin polymerization in cultured human lung cancer cells
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DOI：
10.1016/j.bcp.2006.05.026
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发表时间：
2006-08-28
影响因子：
5.8
通讯作者：
Jia, Hong-Ti
中科院分区：
文献类型：
--
作者：
Gu, Yan-Yan;Zhang, Hong-Yu;Jia, Hong-Ti
肌动蛋白的一个关键特征是它能结合和降解ATP。8-氯腺苷(8-Cl-ADO)在活细胞中可被磷酸化为8-Cl-ATP部分，可抑制肿瘤细胞的增殖。因此，我们验证了8-氯-腺苷可以干扰肌动蛋白聚合的动态的假说。我们发现8-氯-腺苷在体外对人肺癌细胞株A549和H1299的生长有抑制作用，并通过荧光激活细胞分选(FACS)将靶细胞阻滞在G2/M期。免疫细胞化学显示，8-氯-腺苷暴露的细胞微丝的正常组织被破坏，并伴有细胞体积缩小和细胞形态改变，靶细胞出现异常的有丝分裂和凋亡。体外光散射实验表明，8-Cl-ATP可直接抑制G-肌动蛋白向F-肌动蛋白的转化。DNase I抑制实验显示，8-氯-腺苷暴露的A549和H1299细胞的G/F-肌动蛋白比值显著高于未暴露细胞的G/F-肌动蛋白比值。综上所述，这些结果表明，8-氯-腺苷暴露可以改变肌动蛋白聚合的动态性质，扰乱肌动蛋白细丝的动态不稳定性或重排能力。因此，我们的数据提示，8-氯-腺苷可能至少部分地通过干扰微丝的动态不稳定性而发挥其细胞毒性，这可能与其抑制细胞增殖和细胞死亡有关。(C)2006 Elsevier Inc.保留所有权利。
A key feature of actin is its ability to bind and hydrolyze ATP. 8-Chloro-adenosine (8-Cl-Ado), which can be phosphorylated to the moiety of 8-Cl-ATP in living cells, inhibits tumor cell proliferation. Therefore we tested the hypothesis that 8-Cl-Ado can interfere with the dynamic state of actin polymerization. We found that 8-Cl-Ado inhibited the growth of human lung cancer cell line A549 and H1299 in culture, and arrested the target cells in G2/M phase evidenced by fluorescence-activated cell sorting (FACS). Immunocytochemistry showed that the normal organization of microfilaments was disrupted in 8-Cl-Ado-exposed cells, which is accompanied by the decrease of cell size and the alteration of cell shape, and by aberrant mitosis and apoptosis in targeted cells. Furthermore, in vitro light scattering assays revealed that 8-Cl-ATP could directly inhibit the transition of G-actin to F-actin. DNase I inhibition assays showed that the G/F-actin ratio, a surrogate marker of actin polymerization status in living cells, was significantly increased in 8-Cl-Ado-exposed A549 and H1299 cells, compared to the G/F-actin ratio in unexposed cells. Taken together, these results indicate that 8-Cl-Ado exposure can alter the dynamic properties of actin polymerization, disrupt the dynamic instability or the rearrangement ability of actin filaments. Therefore, our data suggest that 8-Cl-Ado may exert its cytotoxicity at least partly by interfering with the dynamic instability of microfilaments, which may correlate with its inhibitory effects on cell proliferation and cell death. (c) 2006 Elsevier Inc. All rights reserved.