MiR-509-3-5p causes aberrant mitosis and anti-proliferative effect by suppression of PLK1 in human lung cancer A549 cells
MiR-509-3-5p causes aberrant mitosis and anti-proliferative effect by suppression of PLK1 in human lung cancer A549 cells
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MiR-509-3-5p 通过抑制人肺癌 A549 细胞中的 PLK1 引起异常有丝分裂和抗增殖作用
DOI：
10.1016/j.bbrc.2016.08.006
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发表时间：
2016
影响因子：
3.1
通讯作者：
Li Shu-Yan
中科院分区：
文献类型：
--
作者：
Wang Xian-Hui;Lu Yao;Liang Jing-Jing;Cao Ji-Xiang;Jin Ya-Qiong;An Guo-Shun;Ni Ju-Hua;Jia Hong-Ti;Li Shu-Yan
microRNA（miRNAs）是基因表达的有效转录后调节因子，在DNA损伤反应（DDR）中发挥作用。PLK 1被鉴定为DNA损伤检查点的调节剂。虽然已经报道了某些microRNA对PLK 1的下调，但对DDR中PLK 1和miR-509-3- 5 p之间的相互作用知之甚少。我们发现miR-509-3- 5 p通过靶向PLK 1的3′-UTR抑制PLK 1的表达，从而导致人肺癌A549细胞的有丝分裂畸变和生长停滞。miR-509-3- 5 p对PLK 1的抑制进一步通过miR-509-3- 5 p在A549、HepG 2和HCT 116 p53 −/−癌细胞中的过表达得到证实，其中PLK 1蛋白被抑制。结果表明，在CIS和ADR诱导的A549细胞中，miR-509-3- 5 p被激活，而PLK 1蛋白表达下调，提示miR-509-3- 5 p对PLK 1的抑制是CIS/ADR诱导的DDR通路的一个组成部分。流式细胞术和免疫荧光标记显示，miR-509-3- 5 p在A549细胞中的过表达可诱导细胞G2/M期阻滞，并导致细胞有丝分裂异常，表现为异常的双极纺锤体、染色体凝聚、滞后DNA和染色体桥。此外，miR-509-3- 5 p的过表达显著地阻断了A549细胞的增殖并且使细胞对CIS和ADR处理敏感。综上所述，miR-509-3- 5 p通过靶向PLK 1是一种可行的癌症抑制剂。我们的数据可能有助于联合化疗的潜在设计，并有助于我们更好地理解microRNA在DNA损伤中的作用。
MicroRNAs (miRNAs) are potent post-transcriptional regulators of gene expression and play roles in DNA damage response (DDR). PLK1 is identified as a modulator of DNA damage checkpoint. Although down-regulation of PLK1 by certain microRNAs has been reported, little is known about the interplay between PLK1 and miR-509-3-5p in DDR. Here we have demonstrated that miR-509-3-5p repressed PLK1 expression by targeting PLK1 3′-UTR, thereby causing mitotic aberration and growth arrest of human lung cancer A549 cells. Repression of PLK1 by miR-509-3-5p was further evidenced by over-expression of miR-509-3-5p in A549, HepG2 and HCT116p53−/−cancer cells, in which PLK1 protein was suppressed. Consistently, miR-509-3-5p was stimulated, while PLK1 protein was down-regulated in A549 cells exposed to CIS and ADR, suggesting that suppression of PLK1 by miR-509-3-5p is a component of CIS/ADR-induced DDR pathway. Flow cytometry and immunofluorescence labeling showed that over-expression of miR-509-3-5p in A549 induced G2/M arrest and aberrant mitosis characterized by abnormal bipolar mitotic spindles, condensed chromosomes, lagging DNA and chromosome bridges. In addition, over-expression of miR-509-3-5p markedly blocked A549 cell proliferation and sensitized the cells to CIS and ADR treatment. Taken together, miR-509-3-5p is a feasible suppressor for cancer by targeting PLK1. Our data may provide aid in potential design of combined chemotherapy and in our better understanding of the roles of microRNAs in response to DNA damage.