'Desperate house genes': the dramatic example of hypoxia.
'Desperate house genes': the dramatic example of hypoxia.
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“绝望的房子基因”：缺氧的戏剧性例子。
DOI：
10.1038/sj.bjc.6605573
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发表时间：
2010-03-16
影响因子：
8.8
通讯作者：
中科院分区：
文献类型：
--
作者：
正常或肿瘤组织和细胞系中的微环境条件可能会干扰进一步的生物学分析。为了仔细评估转录变异，通常使用稳定管家基因（HKG）来标准化定量微阵列或实时聚合酶链反应结果。然而，最近的研究认为，HKG根据组织和治疗而波动。因此，作为在癌症领域常见的一系列条件下HKG变异的一个例子，我们评估缺氧是否会对HKG表达产生影响。在用四种氧浓度（从1至20%）处理的四种细胞系上测量10种常用HKG的表达。在低氧条件下观察到HKG转录物的大变化，并且随着细胞系和氧浓度的不同而沿着不同。为了选择最稳定的HKG，我们比较了基于PCR循环阈值变异系数计算或两种专门的专用软件的三种统计平均值。然而，根据所使用的统计方法，最好的HKG显着不同。此外，使用作为参考，绝对定量的靶基因（这里的蛋白酶激活受体基因1（PAR1）基因），我们表明，有关PAR1的变化，在缺氧的结论可以完全分歧，根据选定的HKG用于标准化。有效HKG的选择将决定将被进一步解释的结果的相关性，因此应该认真考虑。我们的研究结果明确证实，HKG的变化必须在任何实验之前，对每种情况进行精确和系统的确定，以获得可靠的标准化结果，在实验设置，已设计。事实上，这种检测设计，功能的所有体外系统，应仔细评估之前，任何扩展到其他实验模型，包括在体内的。
Microenvironmental conditions in normal or tumour tissues and cell lines may interfere on further biological analysis. To evaluate transcript variations carefully, it is common to use stable housekeeping genes (HKG) to normalise quantitative microarrays or real-time polymerase chain reaction results. However, recent studies argue that HKG fluctuate according to tissues and treatments. So, as an example of HKG variation under an array of conditions that are common in the cancer field, we evaluate whether hypoxia could have an impact on HKG expression. Expression of 10 commonly used HKG was measured on four cell lines treated with four oxygen concentrations (from 1 to 20%). Large variations of HKG transcripts were observed in hypoxic conditions and differ along with the cell line and the oxygen concentration. To elect the most stable HKG, we compared the three statistical means based either on PCR cycle threshold coefficient of variation calculation or two specifically dedicated software. Nevertheless, the best HKG dramatically differs according to the statistical method used. Moreover, using, as a reference, absolute quantification of a target gene (here the proteinase activating receptor gene 1 (PAR1) gene), we show that the conclusions raised about PAR1 variation in hypoxia can totally diverge according to the selected HKG used for normalisation. The choice of a valid HKG will determine the relevance of the results that will be further interpreted, and so it should be seriously considered. The results of our study confirm unambiguously that HKG variations must be precisely and systematically determined before any experiment for each situation, to obtain reliable normalised results in the experimental setting that has been designed. Indeed, such assay design, functional for all in vitro systems, should be carefully evaluated before any extension to other experimental models including in vivo ones.
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影响因子：
8.4
作者：
Naldini, Antonella;Filippi, Irene;Carraro, Fabio
通讯作者：
Carraro, Fabio
鉴定乳腺癌中实时RT-PCR数据归一化的基因。
影响因子：
3.8
作者：
Lyng MB;Laenkholm AV;Pallisgaard N;Ditzel HJ
通讯作者：
Ditzel HJ
影响因子：
2.7
作者：
Aerts, JL;Gonzales, MI;Topalian, SL
通讯作者：
Topalian, SL
影响因子：
2.7
作者：
Dheda, K;Huggett, JF;Zumla, A
通讯作者：
Zumla, A
影响因子：
11.2
作者：
Helczynska, Karolina;Larsson, Anna-Maria;Jirstrom, Karin
通讯作者：
Jirstrom, Karin