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用于人类生长激素的催乳激素反应元件报告基因激活测定

Lactogenic hormone responsive element reporter gene activation assay for human growth hormone.

作者信息

Sakatani T, Kaji H, Takahashi Y, Iida K, Okimura Y, Chihara K

机构信息

Division of Endocrinogy/Metabolism, Neurology and Hematology/Oncology, Department of Clinical Molecular Medicine, Kobe University Graduate School of Medicine, Kobe 650-0017, Japan.

出版信息

Growth Horm IGF Res. 2003 Oct;13(5):275-81. doi: 10.1016/s1096-6374(03)00033-9.

Abstract

We investigated the bioactivity of GH and compared with their immunoactivity in GH bioassay system using lactogenic hormone responsive element (LHRE) reporter gene in Chinese hamster ovary cells transiently co-transfected with human GH receptor cDNA and LHRE/TK-luciferase reporter gene (LHRE/Luc). The recombinant and serum GH but not prolactin almost equally were able to induce LHRE/Luc in a significant and dose-dependent manner, which were equally suppressed by anti-GH. Recombinant GH binding protein (GHBP) at 100 ng/ml but not at 20 ng/ml slightly attenuated GH-induced LHRE/Luc. The serum GH bioactivity (ng/ml) in patients with acromegaly were equal near to their immunoactivity, whereas the bioactivity of the serum GH in a short child with mutant GH (R77C) revealed lower than their immunoactivity. The bioactivity of the recombinant mutant GH was as half as that of wild type GH, thus confirming an antagonistic property of mutant GH. LHRE reporter gene activation assay is useful to measure the GH bioactivity in addition to the conventional bioassay using cell proliferation.

摘要

我们在中国仓鼠卵巢细胞中瞬时共转染人生长激素受体cDNA和促乳素反应元件(LHRE)/胸苷激酶 - 荧光素酶报告基因(LHRE/Luc),利用该促乳素反应元件报告基因系统研究了生长激素(GH)的生物活性,并将其与生长激素生物测定系统中的免疫活性进行比较。重组生长激素和血清生长激素而非催乳素几乎同样能够以显著的剂量依赖性方式诱导LHRE/Luc,且二者均被抗生长激素同样抑制。100 ng/ml而非20 ng/ml的重组生长激素结合蛋白(GHBP)略微减弱生长激素诱导的LHRE/Luc。肢端肥大症患者血清生长激素的生物活性(ng/ml)与其免疫活性相近,而一名携带突变生长激素(R77C)的矮小儿童血清生长激素的生物活性低于其免疫活性。重组突变生长激素的生物活性仅为野生型生长激素的一半,从而证实了突变生长激素具有拮抗特性。除了使用细胞增殖的传统生物测定法外,LHRE报告基因激活测定法对于测量生长激素的生物活性也很有用。

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