Mechanisms of oxidative stress and alterations in gene expression by Libby six-mix in human mesothelial cells.
Mechanisms of oxidative stress and alterations in gene expression by Libby six-mix in human mesothelial cells.
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DOI：
10.1186/1743-8977-7-26
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发表时间：
2010-09-11
影响因子：
10
通讯作者：
Mossman BT
中科院分区：
文献类型：
--
作者：
Hillegass JM;Shukla A;MacPherson MB;Lathrop SA;Alexeeva V;Perkins TN;van der Vliet A;Vacek PM;Gunter ME;Mossman BT
暴露于蒙大拿州利比的闪石纤维导致恶性间皮瘤（MM）的增加，这是一种预后不良的胸膜和腹膜腔肿瘤。使用Affyssin微阵列/GeneSifter分析确定无毒浓度（15×106 μm2/cm 2）的未处理Libby六混合物以及阴性（玻璃珠）和阳性（青石棉）对照对人间皮细胞系（LP 9/TERT-1）基因表达的改变。由于锰超氧化物歧化酶（MnSOD; SOD 2）是唯一在8和24 h显著上调（p < 0.05）的基因，因此我们测定了SOD蛋白和活性、氧化应激和谷胱甘肽（GSH）水平，以更好地了解暴露于无毒（15×106 μm2/cm 2）和毒性浓度（75×106 μm2/cm 2）的Libby six mix后的氧化事件。暴露于15×106 μm2/cm 2 Libby六种混合物在8 h时引起一种基因（SOD 2; 4倍）显著上调（p < 0.05），在24 h时引起111种基因变化，包括SOD 2增加5倍。通过qRT-PCR也证实了HKNM-2正常人胸膜间皮细胞中24小时SOD 2 mRNA水平的增加。结果显示，在75×106 μm2/cm 2的毒性浓度下，24 h SOD 2蛋白水平升高。此外，铜锌超氧化物歧化酶（Cu/ZnSOD; SOD 1）蛋白的水平在24小时在所有矿物质组增加。观察到SOD 2活性呈剂量相关性增加，但总SOD活性保持不变。二氯二氢荧光素二乙酸酯（DCFDA）荧光染色和流式细胞术显示，暴露于Libby六混合物的LP 9/TERT-1细胞的活性氧（ROS）产生呈剂量和时间依赖性增加。Libby six mix和青石棉在75×106 μm2/cm 2时均可引起LPS 9/TERT-1和HKNM-2细胞中GSH的短暂降低（p < 0.05），持续24 h，并增加血红素加氧酶1（HO-1）的基因表达。Libby six-mix导致LP 9/TERT-1人类间皮细胞中的多个基因表达变化，以及SOD 2增加，氧化剂产生增加和细胞内GSH瞬时减少。在相同表面积浓度的无毒玻璃珠中未观察到这些事件。结果支持SOD 2在间皮瘤细胞增殖和凋亡中的重要性及其作为间皮瘤矿物质早期反应的生物标志物的潜在用途的机制基础。
Exposures to an amphibole fiber in Libby, Montana cause increases in malignant mesothelioma (MM), a tumor of the pleural and peritoneal cavities with a poor prognosis. Affymetrix microarray/GeneSifter analysis was used to determine alterations in gene expression of a human mesothelial cell line (LP9/TERT-1) by a non-toxic concentration (15×106 μm2/cm2) of unprocessed Libby six-mix and negative (glass beads) and positive (crocidolite asbestos) controls. Because manganese superoxide dismutase (MnSOD; SOD2) was the only gene upregulated significantly (p < 0.05) at both 8 and 24 h, we measured SOD protein and activity, oxidative stress and glutathione (GSH) levels to better understand oxidative events after exposure to non-toxic (15×106 μm2/cm2) and toxic concentrations (75×106 μm2/cm2) of Libby six-mix. Exposure to 15×106 μm2/cm2 Libby six-mix elicited significant (p < 0.05) upregulation of one gene (SOD2; 4-fold) at 8 h and 111 gene changes at 24 h, including a 5-fold increase in SOD2. Increased levels of SOD2 mRNA at 24 h were also confirmed in HKNM-2 normal human pleural mesothelial cells by qRT-PCR. SOD2 protein levels were increased at toxic concentrations (75×106 μm2/cm2) of Libby six-mix at 24 h. In addition, levels of copper-zinc superoxide dismutase (Cu/ZnSOD; SOD1) protein were increased at 24 h in all mineral groups. A dose-related increase in SOD2 activity was observed, although total SOD activity remained unchanged. Dichlorodihydrofluorescein diacetate (DCFDA) fluorescence staining and flow cytometry revealed a dose- and time-dependent increase in reactive oxygen species (ROS) production by LP9/TERT-1 cells exposed to Libby six-mix. Both Libby six-mix and crocidolite asbestos at 75×106 μm2/cm2 caused transient decreases (p < 0.05) in GSH for up to 24 h and increases in gene expression of heme oxygenase 1 (HO-1) in LP9/TERT-1 and HKNM-2 cells. Libby six-mix causes multiple gene expression changes in LP9/TERT-1 human mesothelial cells, as well as increases in SOD2, increased production of oxidants, and transient decreases in intracellular GSH. These events are not observed at equal surface area concentrations of nontoxic glass beads. Results support a mechanistic basis for the importance of SOD2 in proliferation and apoptosis of mesothelial cells and its potential use as a biomarker of early responses to mesotheliomagenic minerals.
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影响因子：
3.8
作者：
Blake, David J.;Bolin, Celeste M.;Pfau, Jean C.
通讯作者：
Pfau, Jean C.
影响因子：
2.1
作者：
Horton, Kevin;Kapil, Vikas;Anderson, Barbara
通讯作者：
Anderson, Barbara
影响因子：
11.2
作者：
Connor, Kip M.;Hempel, Nadine;Melendez, J. Andres
通讯作者：
Melendez, J. Andres
影响因子：
15.9
作者：
BOYLAN, AM;RUEGG, C;BROADDUS, VC
通讯作者：
BROADDUS, VC
DOI：
10.1165/ajrcmb.10.3.8117443
发表时间：
1994-03-01
影响因子：
6.4
作者：
GRIFFITH, DE;MILLER, EJ;JOHNSON, AR
通讯作者：
JOHNSON, AR