Effect of dihydroartemisinin on UHRF1 gene expression in human prostate cancer PC-3 cells
Effect of dihydroartemisinin on UHRF1 gene expression in human prostate cancer PC-3 cells
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DOI：
10.1097/cad.0000000000000469
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发表时间：
2017-04-01
影响因子：
2.3
通讯作者：
Luo, Ziguo
中科院分区：
文献类型：
--
作者：
Du, Shijuan;Xu, Ge;Luo, Ziguo
前列腺癌作为世界上男性第二常见的癌症，正日益受到人们的关注。双氢青蒿素(DHA)在体内和体外都被证明是一种很有前途的抗癌药物。然而，DHA如何在人前列腺癌PC-3细胞中发挥作用的详细机制仍然不清楚。本研究旨在探讨新型抗癌药物DHA通过抑制PC-3细胞泛素样蛋白PHD和环指蛋白1(Uhrf1)的表达而发挥的作用。流式细胞仪检测细胞凋亡率和细胞周期分布。实时定量聚合酶链式反应检测uhrf1和DNA甲基转移酶1(DNMT1)在mRNA水平的表达，蛋白质印迹法检测uhrf1、DNMT1和p16(Ink4a)蛋白的表达。亚硫酸氢盐基因组测序检测p16(Ink4a)CpG岛的甲基化水平。我们发现DHA诱导PC-3细胞uhrf1和DNMT1表达下调，同时p16(Ink4a)表达上调。在PC-3细胞中，DHA处理组的p16(Ink4a)启动子甲基化水平也降低。此外，DHA还能显著诱导PC-3细胞的凋亡和G1/细胞周期停滞。我们的结果表明，uhrf1/DNMT1的下调是许多细胞事件的上游，包括G1细胞停滞、p16(Ink4a)去甲基化和细胞凋亡。总之，我们的研究为DHA可能作为一种潜在的前列腺癌治疗剂提供了新的证据。版权所有(C)2017 Wolters KluwerHealth，Inc.保留所有权利。
As the second most common cancer in men around the world, prostate cancer is increasingly gaining more attention. Dihydroartemisinin (DHA) has been proven to be a promising anticancer agent in vitro as well as in vivo in accumulating data. However, the detailed mechanisms of how DHA action in human prostate cancer PC-3 cells remain elusive. This study aimed to investigate the effects of DHA, a novel anticancer agent, by inhibiting the expression of ubiquitin like containing PHD and ring finger 1 (UHRF1) in PC-3 cells. The apoptosis and cell-cycle distribution were detected by flow cytometry. Quantitative real-time PCR was performed to examine both UHRF1 and DNA methyltransferase 1 ( DNMT1) expressions at mRNA levels, whereas the expressions of UHRF1, DNMT1, and p16(INK4A) proteins at protein levels were detected by Western blotting. Methylation levels of p16(INK4A) CpG islands were determined by bisulfite genomic sequencing. We showed that DHA induced the downregulation of UHRF1 and DNMT1, accompanied by an upregulation of p16(INK4A) in PC-3 cells. Decreased p16(INK4A) promoter methylation levels in DHA-treated groups were also observed in PC-3 cells. Furthermore, DHA significantly induced apoptosis and G1/cell-cycle arrest in PC-3 cells. Our results suggested that downregulation of UHRF1/DNMT1 is upstream to many cellular events, including G1 cell arrest, demethylation of p16(INK4A), and apoptosis. Together, our study provides new evidence that DHA may serve as a potential therapeutic agent in the treatment of prostate cancer. Copyright (C) 2017 Wolters Kluwer Health, Inc. All rights reserved.