4-hydroxynonenal modifies the effects of serum growth factors on the expression of the c-fos proto-oncogene and the proliferation of HeLa carcinoma cells
4-hydroxynonenal modifies the effects of serum growth factors on the expression of the c-fos proto-oncogene and the proliferation of HeLa carcinoma cells
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DOI：
10.1016/s0891-5849(98)00029-x
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发表时间：
1998-07-01
影响因子：
7.4
通讯作者：
Schaur, RJ
中科院分区：
文献类型：
--
作者：
Kreuzer, T;Grube, R;Schaur, RJ
在这项研究中，4-羟基壬烯醛（HNE），ω-6-多不饱和脂肪酸的过氧化产物，对c-fos原癌基因的表达和生长因子诱导的HeLa癌细胞在体外增殖的影响进行了研究。Fos蛋白与Jun蛋白形成异二聚体AP-1，并通过诱导细胞周期蛋白D1来调节细胞周期。能够诱导c-fos的试剂包括血清、血小板衍生生长因子（PDGF）和表皮生长因子（EGF），所有这些都用于本研究。通过细胞计数（根据台盼蓝排除法的活细胞和死细胞）和BrdU测定来确定增殖率。用逆转录聚合酶链反应（RT-PCR）检测c-fos mRNA水平。在没有HNE的情况下，血清剥夺细胞对血清刺激的反应是c-fos mRNA水平增加10倍以上，DNA合成和细胞增殖速率增加。EGF和PDGF（与胰岛素联合应用）都能够替代FCS，并诱导在无血清培养基中预孵育的肿瘤细胞快速生长。在没有生长因子的情况下，观察到HNE浓度（范围：1-250 μ M）和c-fos mRNA水平之间的负相关。我们假设，在这种情况下，HNE干扰的基础活性的c-fos启动子，EGF，当应用后的HNE治疗，诱导快速生长的肿瘤细胞预孵育在无血清培养基中，如果HNE是在一个生理浓度（1 μ M）。与不含HNE的对照组相比，没有观察到差异，c-fos mRNA水平几乎没有变化。相反，细胞毒性浓度的醛（100 μ M）引起的增殖的完全抑制，虽然在醛处理后立即观察到c-fos mRNA水平的两倍增加。当细胞在PDGF而不是EGF存在下生长时，观察到细胞毒性浓度的HNE对c-fos表达的类似作用。因此，在这两种情况下，HNE可能干扰信号转导途径，这是由外部生长因子启动。c-fos表达的增加可能是一个失败的尝试，以克服由细胞毒性浓度的HNE引起的应激条件的一部分。(C)1998年爱思唯尔科学公司
In this study, the effect of 4-hydroxynonenal (HNE), a peroxidation product of omega-6-poly-unsaturated fatty acids, on the expression of the c-fos proto-oncogene and growth factor-induced proliferation of HeLa carcinoma cells in vitro was investigated. The Fos protein forms the heterodimer AP-1 with the Jun protein and regulates the cell cycle by inducing cyclin D1. Agents that are able to induce c-fos include serum, platelet-derived growth factor (PDGF), and epidermal growth factor (EGF), all of which were used in this study. The proliferation rate was determined by cell counting (viable and dead cells according to trypan blue exclusion) and the BrdU assay. The c-fos mRNA level was monitored by the reverse transcriptase/polymerase chain reaction. In the absence of HNE, serum-deprived cells responded to serum stimulation with a more than l0-fold increase of the c-fos mRNA level as well as with an increased rate of DNA synthesis and cell multiplication. Both EGF and PDGF (applied in combination with insulin) were able to substitute for FCS and induced rapid growth of the tumor cells preincubated in serum-deprived medium. In the absence of growth factors a negative correlation between the HNE concentration (range: 1-250 mu M) and the c-fos mRNA level was observed. We suppose that HNE interferes in this case with the basal activity of the c-fos promoter, EGF, when applied after the HNE treatment, induced rapid growth of the tumor cells preincubated in serum-free medium, if HNE was used in a physiological concentration (1 mu M). NO difference was observed compared to the HNE-free control, c-fos mRNA level was nearly unchanged. In contrast, a cytotoxic concentration of the aldehyde (100 mu M) caused a complete inhibition of proliferation, although a twofold increase of the c-fos mRNA level immediately after the aldehyde treatment was observed. A similar effect of HNE in cytotoxic concentration on c-fos expression was observed when cells were grown in presence of PDGF instead of EGF. Hence, in both cases HNE possibly interferes with the signal transduction pathway, which is initiated by external growth factors. The increased c-fos expression might be part of an abortive attempt to overcome the stressful condition raised by a cytotoxic concentration of HNE. (C) 1998 Elsevier Science Inc.