Poly(ADP-ribosyl)ation enhances H-RAS protein stability and causes abnormal cell cycle progression in human TK6 lymphoblastoid cells treated with hydroquinone
Poly(ADP-ribosyl)ation enhances H-RAS protein stability and causes abnormal cell cycle progression in human TK6 lymphoblastoid cells treated with hydroquinone
复制标题
聚（ADP-核糖基）化可增强 H-RAS 蛋白稳定性，并导致用氢醌处理的人 TK6 淋巴母细胞中细胞周期进程异常。
DOI：
10.1016/j.cbi.2015.05.019
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发表时间：
2015-08-05
影响因子：
5.1
通讯作者：
Zou, Fei
中科院分区：
文献类型：
--
作者：
Liu, Linhua;Ling, Xiaoxuan;Zou, Fei
对苯二酚（Hydroquinone，HQ）是苯的重要代谢产物之一，可诱导细胞周期进程异常，但其诱导机制尚不清楚。聚（ADP-核糖基）化（PAR化）主要由聚（ADP-核糖）聚合酶-1（PARP-1）催化，参与各种生物学过程，包括细胞周期控制。本研究的结果显示，与PBS处理的细胞相比，HQ处理48 h的TK 6人淋巴母细胞在G1期与S期的积累; HQ处理72 h后，细胞从G1期阻滞过渡到S期阻滞。我们检测了与细胞周期或白血病相关的六个基因的表达，以进一步探索这种现象的原因。在这些基因中，发现H-RAS与这一现象相关，因为其mRNA和蛋白表达在48 h时降低，在72 h时升高。PARP活性诱导和抑制实验表明，观察到的PAR化与H-RAS表达呈正相关。此外，在用HQ结合PARP-1敲低处理的细胞中，H-RAS蛋白的表达降低，G1期细胞的数量增加。HQ处理72 h的细胞中H-RAS蛋白的聚（ADP-核糖基）修饰程度增加，进一步支持PAR化的变化导致H-RAS蛋白表达的快速改变，随后是细胞周期的异常进展。采用免疫共沉淀（co-IP）测定来确定PARP-1和H-RAS蛋白是否形成蛋白复合物，并且这些蛋白之间的直接相互作用表明PAR化调节H-RAS表达。激光共聚焦显微镜下可见H-RAS蛋白分布于细胞核和细胞质中。据我们所知，本研究首次揭示了H-RAS蛋白可以通过PAR化修饰。(C)由Elsevier爱尔兰有限公司出版。
Hydroquinone (HQ), one of the most important benzene-derived metabolites, can induce aberrant cell cycle progression; however, the mechanism of this induction remains unclear. Poly(ADP-ribosyl)ation (PARylation), which is catalysed primarily by poly(ADP-ribose) polymerase-1 (PARP-1), participates in various biological processes, including cell cycle control. The results of the present study show an accumulation in G1 phase versus S phase of TK6 human lymphoblast cells treated with HQ for 48 h compared with PBS-treated cells; after 72 h of HQ treatment, the cells transitioned from G1 arrest to S phase arrest. We examined the expression of six genes related to the cell cycle or leukaemia to further explore the reason for this phenomenon. Among these genes, H-RAS was found to be associated with this phenomenon because its mRNA and protein expression decreased at 48 h and increased at 72 h. Experiments for PARP activity induction and inhibition revealed that the observed PARylation was positively associated with H-RAS expression. Moreover, in cells treated with HQ in conjunction with PARP-1 knockdown, expression of the H-RAS protein decreased and the number of cells in G1 phase increased. The degree of poly(ADP-ribosyl) modification of the H-RAS protein increased in cells treated with HQ for 72 h, further supporting that changes in PARylation contributed to the rapid alteration of H-RAS protein expression, followed by abnormal progression of the cell cycle. Co-immunoprecipitation (co-IP) assays were employed to determine whether protein complexes were formed by PARP-1 and H-RAS proteins, and the direct interaction between these proteins indicated that PARylation regulated H-RAS expression. As detected by confocal microscopy, the H-RAS protein was found in the nucleus and cytoplasm. To our knowledge, this study is the first to reveal that H-RAS protein can be modified by PARylation. (C) 2015 Published by Elsevier Ireland Ltd.