Decoding NADPH oxidase 4 expression in human tumors.
Decoding NADPH oxidase 4 expression in human tumors.
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DOI：
10.1016/j.redox.2017.05.016
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发表时间：
2017-10
期刊：
影响因子：
11.4
通讯作者：
Doroshow JH
中科院分区：
文献类型：
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作者：
Meitzler JL;Makhlouf HR;Antony S;Wu Y;Butcher D;Jiang G;Juhasz A;Lu J;Dahan I;Jansen-Dürr P;Pircher H;Shah AM;Roy K;Doroshow JH
NADPH氧化酶4(NOX4)是一种氧化还原活性的膜相关蛋白，通过其结构性产生过氧化氢的能力，在人类癌症中起到基因组不稳定、氧化还原信号和辐射敏感性的作用。大多数关于NOX4在恶性肿瘤中的研究集中于对少数肿瘤细胞系的评估，而不是对人类肿瘤标本本身的评估；此外，这些研究往往使用了尚未得到很好表征的免疫学工具。为了确定NOX4在广泛的实体肿瘤中的表达情况，我们开发了一种新型的单抗，它可以识别人类NOX4蛋白的一个特定的胞外区域，并且不与NOX基因家族的其他六个成员中的任何一个发生交叉反应。对20组上皮性肿瘤的评估首次发现NOX4在头颈部癌(15/19例)、食道癌(12/18例)、膀胱癌(10/19例)、卵巢癌(6/17例)、前列腺癌(7/19例)以及恶性黑色素瘤(7/15例)中高表达。检测到低水平的转化生长因子-β1对NOX4蛋白表达的上调，证明了该新探针的敏感性；免疫荧光实验发现，卵巢癌细胞内源性NOX4的高水平表达仅与核周膜有关。这些研究表明，与正常组织相比，在明确定义的特定人类癌症组中，NOX4的表达上调，并且它的表达以一种可能调节DNA氧化损伤的方式定位在细胞膜上。NADPH氧化酶4(NOX4)的检测是用一种针对胞外E环区(氨基酸209-282)的单抗(47-6)实现的。对20例人类恶性肿瘤的组织芯片分析显示，IHC检测的NOX4在膀胱癌、食道癌、头颈部、卵巢癌、前列腺癌和黑色素瘤中的差异表达高于正常组织。NOX4在卵巢癌细胞系COV362中的表达定位于核周。
NADPH oxidase 4 (NOX4) is a redox active, membrane-associated protein that contributes to genomic instability, redox signaling, and radiation sensitivity in human cancers based on its capacity to generate H2O2 constitutively. Most studies of NOX4 in malignancy have focused on the evaluation of a small number of tumor cell lines and not on human tumor specimens themselves; furthermore, these studies have often employed immunological tools that have not been well characterized. To determine the prevalence of NOX4 expression across a broad range of solid tumors, we developed a novel monoclonal antibody that recognizes a specific extracellular region of the human NOX4 protein, and that does not cross-react with any of the other six members of the NOX gene family. Evaluation of 20 sets of epithelial tumors revealed, for the first time, high levels of NOX4 expression in carcinomas of the head and neck (15/19 patients), esophagus (12/18 patients), bladder (10/19 patients), ovary (6/17 patients), and prostate (7/19 patients), as well as malignant melanoma (7/15 patients) when these tumors were compared to histologically-uninvolved specimens from the same organs. Detection of NOX4 protein upregulation by low levels of TGF-β1 demonstrated the sensitivity of this new probe; and immunofluorescence experiments found that high levels of endogenous NOX4 expression in ovarian cancer cells were only demonstrable associated with perinuclear membranes. These studies suggest that NOX4 expression is upregulated, compared to normal tissues, in a well-defined, and specific group of human carcinomas, and that its expression is localized on intracellular membranes in a fashion that could modulate oxidative DNA damage. NADPH oxidase 4 (NOX4) detection has been achieved with a monoclonal antibody (47-6) developed to the extracellular E-loop region (amino acids 209–282). Tissue microarray analysis of 20 human malignancies reveals differential NOX4 expression by IHC was higher in bladder, esophageal, head and neck, ovary, and prostate cancers and melanoma than in normal tissues. NOX4 expression in the ovarian cancer cell line COV362 is localized in the perinuclear region.