Effects of JS-K, a novel anti-cancer nitric oxide prodrug, on gene expression in human hepatoma Hep3B cells
Effects of JS-K, a novel anti-cancer nitric oxide prodrug, on gene expression in human hepatoma Hep3B cells
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新型抗癌一氧化氮前药JS-K对人肝癌Hep3B细胞基因表达的影响
DOI：
10.1016/j.biopha.2017.01.080
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发表时间：
2017-04-01
影响因子：
7.5
通讯作者：
Saavedra, Joseph E.
中科院分区：
文献类型：
--
作者：
Dong, Ray;Wang, Xueqian;Saavedra, Joseph E.
JS-K是一种新型的抗癌一氧化氮（NO）前药，对多种癌细胞有效，包括抑制大鼠AM-1肝癌细胞生长。为了进一步评价JS-K的抗癌作用，用不同浓度（0-100 μ M）的JS-K和化合物对照JS-43-126处理人肝癌Hep 3B细胞24小时，并通过MTS测定法测定细胞毒性。化合物对照JS-43-126在高达100 μ M的浓度下对Hep 3B细胞没有细胞毒性，而JS-K的LC 50约为10 mM。为了检查JS-K的抗肿瘤作用的分子机制，用1-10 mM的JS-K处理Hep 3B细胞24小时，然后通过真实的时间RT-PCR进行基因表达分析，并通过共聚焦图像进行蛋白质免疫染色。JS-K是靶向NO的GST-α前药，并且GST-α的免疫染色减少与JS-K治疗相关。JS-K激活Hep 3B细胞的凋亡途径，包括诱导caspase-3、caspase-9、Bax、TNF-α和IL-1 β，并且caspase-3的免疫染色增强。JS-K在转录水平和蛋白水平均能上调血小板反应蛋白-1（TSP-1）和金属蛋白酶组织抑制因子-1（TIMP-1）的表达。JS-K处理后，Hep 3B细胞分化相关基因CD 14、CD 11b表达增加，c-myc表达降低。因此，多个分子事件似乎与JS-K在人肝癌Hep 3B细胞中的抗癌作用相关，包括与凋亡相关的基因的激活和参与抗血管生成和肿瘤细胞迁移的基因的诱导。(C)2017 Elsevier Masson SAS。All rights reserved.
JS-K is a novel anticancer nitric oxide (NO) prodrug effective against a variety of cancer cells, including the inhibition of AM-1 hepatoma cell growth in rats. To further evaluate anticancer effects of JS-K, human hepatoma Hep3B cells were treated with JS-K and the compound control JS-43-126 at various concentrations (0-100 mu M) for 24 h, and cytotoxicity was determined by the MTS assay. The compound control JS-43-126 was not cytotoxic to Hep3B cells at concentrations up to 100 mu M, while the LC50 for JS-K was about 10 mM. To examine the molecular mechanisms of antitumor effects of JS-K, Hep3B cells were treated with 1-10 mM of JS-K for 24 h, and then subjected to gene expression analysis via real time RT-PCR and protein immunostain via confocal images. JS-K is a GST-alpha targeting NO prodrug, and decreased immunostaining for GST-a was associated with JS-K treatment. JS-K activated apoptosis pathways in Hep3B cells, including induction of caspase-3, caspase-9, Bax, TNF-alpha, and IL-1 beta, and immunostaining for caspase-3 was intensified. The expressions of thrombospondin-1 (TSP-1) and the tissue inhibitors of metalloproteinase-1 (TIMP-1) were increased by JS-K at both transcript and protein levels. JS-K treatment also increased the expression of differentiation-related genes CD14 and CD11b, and depressed the expression of c-myc in Hep3B cells. Thus, multiple molecular events appear to be associated with anticancer effects of JS-K in human hepatoma Hep3B cells, including activation of genes related to apoptosis and induction of genes involved in antiangiogenesis and tumor cell migration. (C) 2017 Elsevier Masson SAS. All rights reserved.