The p53-inhibitor pifithrin-alpha inhibits firefly luciferase activity in vivo and in vitro.
The p53-inhibitor pifithrin-alpha inhibits firefly luciferase activity in vivo and in vitro.
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DOI：
10.1186/1471-2199-4-9
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发表时间：
2003-09-11
影响因子：
--
通讯作者：
Perkins ND
中科院分区：
文献类型：
--
作者：
Rocha S;Campbell KJ;Roche KC;Perkins ND
倍氟菊酯-α是P53转录活性的小分子抑制物。有人建议，在癌症的化疗和放射治疗中联合使用Pifithin-α将减少这些治疗在正常组织中的副作用，这些组织中仍然含有野生型p53。此外，Pifithin-α在实验室中为研究模型系统中P53的功能提供了一个有用的工具。在研究Pifithin-α对NF-κB转录活性的影响时，我们观察到含有萤火虫荧光素酶基因的报告质粒具有很强的抑制作用。萤火虫荧光素酶是报告基因分析中最常用的酶之一。相反，含有雷尼拉荧光素酶或氯霉素乙酰转移酶的报告质粒未观察到抑制作用。在体内和体外观察了α对萤火虫荧光素酶活性的抑制作用。α不能抑制萤火虫荧光素酶蛋白的表达，而是抑制光的产生/发射，因为在活性提取物中加入外源α抑制了这一活性。另外，α还能抑制重组萤火虫荧光素酶蛋白的活性。在它的其他生物活性中，Pifithin-α是萤火虫荧光素酶活性的抑制剂。因此，当使用这种化合物时，必须小心，这种化合物被描述为P53转录活性的抑制物，使用瞬时转染的报告质粒来研究对基因表达的影响。此外，这些结果表明，当使用新的化合物时，实验过程中使用的载体的选择对于得出正确的结论可能是非常重要的。
Pifithrin-α is a small molecule inhibitor of p53 transcriptional activity. It has been proposed that the use of pifithrin-α in conjunction with chemotherapeutic and radiation therapies for cancer will reduce the side effects of these treatments in normal tissue that still contains wild type p53. In addition, pifithrin-α provides a useful tool in the laboratory to investigate the function of p53 in model systems. While investigating the effects of pifithrin-α on the transcriptional activity of NF-κB, we observed a strong inhibition of reporter plasmids containing the firefly luciferase gene. Firefly luciferase is one of the most commonly used enzymes in reporter gene assays. In contrast, no inhibition of reporter plasmids containing Renilla luciferase or chloramphenicol acetyltransferase was observed. The inhibition of firefly luciferase activity by pifithrin-α was observed both in vivo and in vitro. Pifithrin-α did not inhibit firefly luciferase protein expression, but rather suppressed light production/emission, since addition of exogenous pifithrin-α to active extracts inhibited this activity. Furthermore, pifithrin-α also inhibited recombinant firefly luciferase protein activity. Among its other biological activities, pifithrin-α is an inhibitor of firefly luciferase activity. Caution must therefore be taken when using this compound, which has been characterised as an inhibitor of p53 transcriptional activity, to investigate effects on gene expression using transiently transfected reporter plasmids. Furthermore, these results demonstrate that when using novel compounds, the choice of vectors used in the experimental procedures might be of great importance for the correct conclusions to be made.
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DOI：
10.1016/s0027-5107(01)00273-1
发表时间：
2002-02-20
影响因子：
2.3
作者：
Bassi, L;Carloni, M;Palitti, F
通讯作者：
Palitti, F
DOI：
10.1073/pnas.88.1.134
发表时间：
1991-01-01
影响因子：
11.1
作者：
MOSS, GWJ;FRANKS, NP;LIEB, WR
通讯作者：
LIEB, WR
影响因子：
5.3
作者：
Rocha, S;Martin, AM;Perkins, ND
通讯作者：
Perkins, ND
影响因子：
3.4
作者：
Franks, NP;Jenkins, A;Brick, P
通讯作者：
Brick, P
影响因子：
8
作者：
Qin, JZ;Chaturvedi, V;Nickoloff, BJ
通讯作者：
Nickoloff, BJ