Expression of reduced nicotinamide adenine dinucleotide phosphate oxidase (ThoX, LNOX, Duox) genes and proteins in human thyroid tissues.
Expression of reduced nicotinamide adenine dinucleotide phosphate oxidase (ThoX, LNOX, Duox) genes and proteins in human thyroid tissues.
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DOI：
10.1210/jcem.86.7.7646
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发表时间：
2001-07
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影响因子：
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通讯作者：
Corinne Dupuy;L. Lacroix;M. Nocera;Monique Talbot;R. Ohayon;D. Dème;Jean-Michel Bidart;Martin S
中科院分区：
文献类型：
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作者：
Corinne Dupuy;L. Lacroix;M. Nocera;Monique Talbot;R. Ohayon;D. Dème;Jean-Michel Bidart;Martin S
NADPH氧化酶黄素蛋白LNOX 2的大型同系物，可能还有LNOX 1，是参与甲状腺H（2）O（2）生成的黄素蛋白。使用抗肽抗体对正常人甲状腺的膜蛋白进行Western印迹分析，结果表明LNOX 1，2是164 kDa的糖蛋白，N-糖基化基序占其总表观分子量的至少10-20 kDa。对23种不同人体组织的北方印迹分析表明，LNOX 2信使RNA（mRNA）仅在甲状腺中强烈表达，尽管表达序列标签数据库的blast分析表明，LNOX基因也在一些非甲状腺细胞中表达。我们分别采用实时定量PCR和抗肽抗体检测了正常和病理甲状腺组织中LNOX 1，2基因和蛋白的表达。在正常组织中，LNOX 1、2定位于甲状腺细胞的顶端。LNOX 1，2的免疫染色是异质的，在给定的卵泡内，有40-60%的阳性卵泡细胞。在正常组织和病理组织中，LNOX 1和LNOX 2 mRNA水平的变化是平行的，这表明两种基因表达的调节相似。而LNOX mRNA似乎在良性疾病中受到轻微影响，蛋白质的表达是高度可变的。在多结节性甲状腺肿中，40-60%的细胞被染色。在功能减退的腺瘤中，LNOX免疫染色在滤泡之间高度可变，而钠/碘（Na+/I-）同向转运体免疫染色减少。在功能亢进的甲状腺组织中，只有少数细胞（0-10%）被弱染色，而钠/碘同向转运体染色在大多数滤泡细胞中发现。总之，LNOX蛋白是一种新的顶端糖蛋白，其表达调控与其他甲状腺标志物不同。
The large homolog of NADPH oxidase flavoprotein LNOX2, and probably LNOX1, are flavoproteins involved in the thyroid H(2)O(2) generator. Western blot analysis of membrane proteins from normal human thyroid, using antipeptide antibodies, indicated that LNOX1,2 are 164-kDa glycoproteins and that N-glycosylated motifs account for at least 10-20 kDa of their total apparent molecular mass. Northern blot analysis of 23 different human tissues demonstrated that LNOX2 messenger RNA (mRNA) is strongly expressed only in the thyroid gland, although blast analysis of expressed sequence tags databases indicated that LNOX genes are also expressed in some nonthyroid cells. We investigated LNOX1,2 gene and protein expressions in normal and pathological human thyroid tissues using real-time kinetic quantitative PCR and antipeptide antibodies, respectively. In normal tissue, LNOX1,2 are localized at the apical pole of thyrocytes. Immunostaining for LNOX1,2 was heterogeneous, inside a given follicle, with 40-60% of positive follicular cells. Among normal and pathological tissues, variations of LNOX1 and LNOX2 mRNA levels were parallel, suggesting a similar regulation of both gene expressions. Whereas LNOX mRNAs seemed slightly affected in benign disease, the expression of protein was highly variable. In multinodular goiters, 40-60% of cells were stained. In hypofunctioning adenomas, LNOX immunostaining was highly variable among follicles, whereas sodium/iodide (Na+/I-) symporter immunostaining was decreased. In hyperfunctioning thyroid tissues, only few cells (0-10%) were weakly stained, whereas sodium/iodide symporter staining was found in the majority of follicular cells. In conclusion, LNOX proteins are new apical glycoproteins with a regulation of expression that differs from other thyroid markers.