A quantitative bioassay for HIV-1 gene expression based on UV activation: effect of glycyrrhizic acid
A quantitative bioassay for HIV-1 gene expression based on UV activation: effect of glycyrrhizic acid
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DOI：
10.1016/j.antiviral.2003.11.005
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发表时间：
2004-04-01
影响因子：
7.6
通讯作者：
Lin, JC
中科院分区：
文献类型：
--
作者：
Cherng, JM;Lin, HJ;Lin, JC
先前的报道表明，在HeLa细胞中稳定转染的HIV-LTRcat构建体在暴露于UV光后是可诱导的。我们优化了该系统，用于研究药物对HIV-1基因表达的影响。最大的UV响应，观察到在静止的静止细胞刺激新鲜培养基为3它。甘草酸抑制UV诱导的HIV基因表达的浓度依赖性的方式。在紫外光照射后立即加入GL，其抑制作用最强;在紫外光照射后5 h加入GL，其抑制作用仍较明显，10 h后完全消失。如果在UV暴露前用亚有效剂量（0.0012 mM）GL预处理细胞，则抑制作用甚至更明显。预处理和未预处理的IC 50值分别为0.04和0.38 mM。GL对UV诱导的CAT活性的抑制作用与NF-κ B p65、NF-κ B p50、c-Fos和c-Rel结合活性的完全抑制相关。因此，本文提出的基于UV的生物测定法可用于干扰HIV-1基因表达的化合物的常规筛选。(C)2003 Elsevier B. V.保留所有权利。
Previous reports have shown that HIV-LTRcat constructs stably transfected in HeLa cells are inducible after exposure to UV light. We have optimized this system for studying the effect of drugs on HIV-1 gene expression. The maximum UV response was observed in quiescent stationary cells stimulated with fresh medium for 3 It. Glycyrrhizic acid suppressed UV-induced HIV gene expression in a concentration-dependent manner. The inhibitory effect was strongest when GL was added immediately after UV exposure; it was still evident when GL was added at 5 h, it was completely lost at 10 h, after UV exposure. The inhibitory effect was even more pronounced if the cells were pretreated with sub-effective dose (0.0012 mM) of GL prior to UV exposure. The IC50 values with and without pretreatment were 0.04 and 0.38 mM, respectively. Cell proliferation and viability were not affected by GL at doses as high as 2.4 mM. The inhibitory effect of GL on UV-induced CAT activity correlated with the complete inhibition of binding activities of NF-kappaB p65, NF-kappaB p50, c-Fos, and c-Rel. Thus, the UV-based bioassay as proposed here can be exploited for the routine screening of the compounds that interfere with HIV-1 gene expression. (C) 2003 Elsevier B.V. All rights reserved.