Overexpression of UHRF1 promoted the proliferation of vascular smooth cells via the regulation of Geminin protein levels
Overexpression of UHRF1 promoted the proliferation of vascular smooth cells via the regulation of Geminin protein levels
复制标题
UHRF1过表达通过调节Geminin蛋白水平促进血管平滑细胞增殖
DOI：
10.1042/bsr20181341
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发表时间：
2019-02-28
影响因子：
4
通讯作者：
Shu, Mao-qin
中科院分区：
文献类型：
--
作者：
Chen, Xia;Zhou, You-li;Shu, Mao-qin
Geminin 是 DNA 复制许可和细胞周期的抑制剂。我们之前的研究表明，Geminin 在调节血管平滑细胞 (VSMC) 的表型多样性和生长中发挥着重要作用。具有 PHD 和环指结构域 1 (UHRF1) 的类泛素是一种表观遗传协调子，其 RING 结构域赋予内在的 E3 连接酶活性，介导多种蛋白质的泛素化和蛋白质-蛋白质相互作用。 UHRF1 的异常表达与多种人类恶性肿瘤的侵袭性有关，其中 UHRF1 的敲低导致癌细胞增殖减少。然而，尚不清楚正确的 UHRF1 功能是否通过改变 Geminin 蛋白水平参与 VSMC 的异常增殖和表型转换。在本研究中，在UHRF1过表达的A10细胞中，使用3H-胸苷和5-乙炔基-20-脱氧尿苷（EdU）和CCK8来检查VSMC的增殖。进行 RT-PCR 和蛋白质印迹分析来研究 UHRF1 介导的效果是否是通过改变 VSMC 中 Geminin 的表达来实现的。进行RNA-seq分析来剖析这些效应的相关机制或信号通路。体外实验结果表明，UHRF1通过下调Geminin蛋白水平促进VSMCs增殖和细胞周期，但Geminin mRNA表达无变化。此外，UHRF1上调后PI3K-Akt信号通路增加。我们的研究表明，过表达 UHRF1 通过降低抑制性 Geminin 蛋白水平来促进细胞周期以及激活 PI3K-Akt 信号传导，从而参与 VSMC 增殖。这可能为制定更好的策略来预防与 VSMC 异常增殖相关的疾病提供关键知识。
Geminin is an inhibitor of DNA replication licensing and cell cycle. Our previous study demonstrates that Geminin plays an important role in regulating phenotypic diversity and growth of vascular smooth cells (VSMCs). Ubiquitin-like with PHD and RING Finger domains 1 (UHRF1) is an epigenetic coordinator, whose RING domain confers intrinsic E3 ligase activity, mediating the ubiquitination of several proteins and the protein–protein interaction. Aberrant expression of UHRF1 was related to aggressiveness of multiple human malignancies, where knockdown of UHRF1 led to decreased proliferation of cancer cells. However, it is unclear whether proper UHRF1 function is involved in aberrant proliferation and phenotypic switching of VSMCs via altering Geminin protein levels. In present study, in UHRF1-overexpressing A10 cells, 3H-thymidine and 5-ethynyl-20-deoxyuridine (EdU) and CCK8 were used to examine the proliferation of VSMCs. RT-PCR and Western blot analyses were performed to investigate whether UHRF1-mediated effects were achieved by altering Geminin expression in VSMCs. RNA-seq analysis was performed to dissect related mechanisms or signaling pathways of these effects. The results of in vitro experiments suggested that UHRF1 prompted proliferation and cell cycle of VSMCs via the down-regulation of Geminin protein levels with no change in Geminin mRNA expression. Besides, PI3K-Akt signaling pathway was increased upon UHRF1 up-regulation. Our study demonstrated that overexpressing UHRF1 was involved in VSMCs proliferation through reducing inhibitory Geminin protein levels to promote cell cycle as well as activating PI3K-Akt signaling. This may provide key knowledge for the development of better strategies to prevent diseases related to VSMCs abnormal proliferation.