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首页> 外文期刊>Clinical and diagnostic laboratory immunology >Enzyme-Linked Immunosorbent Assay for Pharmacological Studies Targeting Hypoxia-Inducible Factor 1α
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Enzyme-Linked Immunosorbent Assay for Pharmacological Studies Targeting Hypoxia-Inducible Factor 1α

机译：酶联免疫吸附法用于针对缺氧诱导因子1α的药理研究

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Hypoxia-inducible factor 1 (HIF-1) activates the transcription of a wide range of genes related to oxygen delivery and metabolic adaptation under hypoxic (low-oxygen) conditions. HIF-1 is, in fact, a heterodimer of two subunits, HIF-1α and HIF-1β. The only analytical methods available for measuring HIF-1α levels in tumors are immunohistochemistry and Western blotting. Immunohistochemistry has the advantage of allowing the identification and direct examination of HIF-1α-expressing cells, but has the intrinsic limitation, as for Western blotting, of being nonquantitative. We developed and validated an enzyme-linked immunosorbent assay (ELISA) approach to measure HIF-1α levels in cultured tumor cell lines in vitro. HIF-1α was expressed in thirteen tumor cell lines grown under hypoxic conditions; however, the levels differed strongly between cell lines. These data point to intrinsic differences between cell lines for the induction of HIF-1α under hypoxic conditions. The ELISA developed in the present study is thus an interesting alternative to other analytical methods used to measure HIF-1α protein levels and should be useful in preclinical pharmacological studies targeting HIF-1α.
机译：缺氧诱导因子1（HIF-1）激活与缺氧（低氧）条件下氧气输送和代谢适应有关的多种基因的转录。实际上，HIF-1是两个亚基HIF-1α和HIF-1β的异二聚体。可用于测量肿瘤中HIF-1α水平的唯一分析方法是免疫组织化学和Western印迹。免疫组织化学的优点是可以鉴定和直接检查表达HIF-1α的细胞，但就蛋白质印迹而言，其固有的局限性是不能定量。我们开发并验证了一种酶联免疫吸附测定（ELISA）方法，用于体外测量培养的肿瘤细胞系中的HIF-1α水平。 HIF-1α在低氧条件下生长的13种肿瘤细胞系中表达。但是，细胞系之间的水平差异很大。这些数据指出了在缺氧条件下诱导HIF-1α的细胞系之间的内在差异。因此，本研究中开发的ELISA是用于测量HIF-1α蛋白水平的其他分析方法的有趣替代方法，并且应在针对HIF-1α的临床前药理研究中有用。

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