Inhibition of hydrogen peroxide signaling by 4-hydroxynonenal due to differential regulation of Akt1 and Akt2 contributes to decreases in cell survival and proliferation in hepatocellular carcinoma cells.
Inhibition of hydrogen peroxide signaling by 4-hydroxynonenal due to differential regulation of Akt1 and Akt2 contributes to decreases in cell survival and proliferation in hepatocellular carcinoma cells.
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DOI：
10.1016/j.freeradbiomed.2012.04.021
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发表时间：
2012-07-01
影响因子：
7.4
通讯作者：
Petersen, Dennis R.
中科院分区：
文献类型：
--
作者：
Shearn, Colin T.;Reigan, Philip;Petersen, Dennis R.
4-羟基壬烯醛 (4-HNE) 等亲电子试剂导致的细胞信号传导失调是慢性炎症性肝病发病机制的关键组成部分。炎症的另一个后果是通过产生活性氧化物质（例如过氧化氢）而导致氧化损伤持续存在。此前，我们已经证明 Akt2 是肝细胞癌细胞中 4-HNE 的直接靶标。在本研究中，我们使用肝细胞癌细胞系 HepG2 作为模型来了解 4-HNE 和过氧化氢的组合效应。我们证明 4-HNE 抑制过氧化氢介导的 Akt1 磷酸化，但不抑制 Akt2。与未处理的对照细胞相比，用 4-HNE 预处理 HepG2 细胞可防止过氧化氢刺激下游靶标的 Akt 依赖性磷酸化和细胞内 Akt 活性。使用生物素酰肼捕获，证实 4-HNE 处理导致 Akt1 羰基化，而在未经处理的对照细胞中未观察到这种情况。使用合成的 GSK3α/β 肽作为底物，用 20μM 或 40μM 4-HNE 处理重组人肉豆蔻酰化 Akt1 (rAkt1)，分别抑制 rAkt1 活性 29% 和 60%。我们进一步证明 4-HNE 通过 PI3 激酶和 PP2A 依赖性机制激活 Erk，导致 Jnk 磷酸化增加。 MTT 测定和 EdU 掺入证明，在较高浓度下，4-HNE 会降低细胞存活和增殖，并降低细胞周期蛋白 D1 和 β-连环蛋白的表达，而添加过氧化氢仅会适度增强这种效果。 4-HNE 对 Erk、Jnk 和 Akt 依赖性细胞存活途径发挥组合作用的能力为了解与慢性炎症相关的细胞损伤机制提供了额外的见解。
Dysregulation of cell signaling by electrophiles such as 4-hydroxynonenal (4-HNE) is a key component in the pathogenesis of chronic inflammatory liver disease. Another consequence of inflammation is the perpetuation of oxidative damage by the production of reactive oxidative species such as hydrogen peroxide. Previously, we have demonstrated Akt2 as a direct target of 4-HNE in hepatocellular carcinoma cells. In the present study, we used the hepatocellular carcinoma cell line HepG2 as model to understand the combinatorial effects of 4-HNE and hydrogen peroxide. We demonstrate that 4-HNE inhibits hydrogen peroxide mediated phosphorylation of Akt1 but not Akt2. Pretreatment of HepG2 cells with 4-HNE prevented hydrogen peroxide stimulation of Akt-dependent phosphorylation of downstream targets and intracellular Akt activity compared with untreated control cells. Using biotin hydrazide capture, it was confirmed that 4-HNE treatment resulted in carbonylation of Akt1, which was not observed in untreated control cells. Using a synthetic GSK3α/β peptide as a substrate, treatment of recombinant human myristoylated Akt1 (rAkt1) with 20μM or 40μM 4-HNE inhibited rAkt1 activity by 29% and 60%, respectively. We further demonstrate that 4-HNE activates Erk via a PI3 kinase and PP2A-dependent mechanism leading to increased Jnk phosphorylation. At higher concentrations, 4-HNE decreased both cell survival and proliferation as evidenced by MTT assays and EdU incorporation as well as decreased expression of cyclin D1 and β-catenin, an effect only moderately increased by the addition of hydrogen peroxide. The ability of 4-HNE to exert combinatorial effects on Erk, Jnk and Akt-dependent cell survival pathways provides additional insight into the mechanisms of cellular damage associated with chronic inflammation.
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DOI：
10.1074/mcp.m800070-mcp200
发表时间：
2009-04
期刊：
Molecular & cellular proteomics : MCP
影响因子：
--
作者：
Codreanu SG;Zhang B;Sobecki SM;Billheimer DD;Liebler DC
通讯作者：
Liebler DC
影响因子：
4.8
作者：
Lee, SR;Yang, KS;Rhee, SG
通讯作者：
Rhee, SG
影响因子：
4.8
作者：
Cheung, Samuel M. S.;Kornelson, Jennifer C.;Marshall, Aaron J.
通讯作者：
Marshall, Aaron J.
影响因子：
4.7
作者：
Lee, TK;Man, K;Fan, ST
通讯作者：
Fan, ST
影响因子：
5.7
作者：
Lim, Ji-Hong;Lee, Yoon-Mi;Park, Jong-Wan
通讯作者：
Park, Jong-Wan