Long non-coding RNA ZFAS1 interacts with CDK1 and is involved in p53-dependent cell cycle control and apoptosis in colorectal cancer.
Long non-coding RNA ZFAS1 interacts with CDK1 and is involved in p53-dependent cell cycle control and apoptosis in colorectal cancer.
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DOI：
10.18632/oncotarget.5807
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发表时间：
2016-01-05
期刊：
影响因子：
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通讯作者：
Slaby O
中科院分区：
文献类型：
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作者：
Thorenoor N;Faltejskova-Vychytilova P;Hombach S;Mlcochova J;Kretz M;Svoboda M;Slaby O
我们确定了83个长非编码RNA（lncRNA）的表达，并确定了ZFAS 1在结直肠癌（CRC）组织中显著上调。在119例结直肠癌患者中，我们观察到111例结直肠癌组织中ZFAS 1的表达至少是配对正常结直肠组织的两倍（P < 0.0001）。通过使用CRC细胞系（HCT 116 +/+，HCT 116 −/−和DLD-1），我们发现ZFAS 1沉默通过细胞周期的G1停滞降低增殖，以及CRC细胞的致瘤性。我们通过下拉实验和RNA免疫沉淀法鉴定了ZFAS 1的相互作用伴侣细胞周期蛋白依赖性激酶1（CDK 1）。此外，我们已经通过生物信息学方法预测ZFAS 1海绵miR-590- 3 p，其被证明靶向CDK 1。CDK 1水平不受ZFAS 1沉默的影响，但细胞周期蛋白B1在两种细胞系中均降低。我们观察到在ZFAS 1沉默后CRC细胞系中p53水平和PARP裂解的显著增加，表明细胞凋亡增加。我们的数据表明，ZFAS 1可能通过两种主要作用在CRC中作为癌基因发挥作用：（i）通过p53的不稳定性和（ii）通过与CDK 1/细胞周期蛋白B1复合物的相互作用，导致细胞周期进展和抑制凋亡。然而，这些相互作用背后的分子机制必须进一步澄清。
We determined expression of 83 long non-coding RNAs (lncRNAs) and identified ZFAS1 to be significantly up-regulated in colorectal cancer (CRC) tissue. In cohort of 119 CRC patients we observed that 111 cases displayed at least two-times higher expression of ZFAS1 in CRC compared to paired normal colorectal tissue (P < 0.0001). By use of CRC cell lines (HCT116+/+, HCT116−/− and DLD-1) we showed, that ZFAS1 silencing decreases proliferation through G1-arrest of cell cycle, and also tumorigenicity of CRC cells. We identified Cyclin-dependent kinase 1 (CDK1) as interacting partner of ZFAS1 by pull-down experiment and RNA immunoprecipitation. Further, we have predicted by bioinformatics approach ZFAS1 to sponge miR-590-3p, which was proved to target CDK1. Levels of CDK1 were not affected by ZFAS1 silencing, but cyclin B1 was decreased in both cell lines. We observed significant increase in p53 levels and PARP cleavage in CRC cell lines after ZFAS1 silencing indicating increase in apoptosis. Our data suggest that ZFAS1 may function as oncogene in CRC by two main actions: (i) via destabilization of p53 and through (ii) interaction with CDK1/cyclin B1 complex leading to cell cycle progression and inhibition of apoptosis. However, molecular mechanisms behind these interactions have to be further clarified.