Identification of suitable endogenous control genes for microRNA gene expression analysis in human breast cancer.
Identification of suitable endogenous control genes for microRNA gene expression analysis in human breast cancer.
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DOI：
10.1186/1471-2199-9-76
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发表时间：
2008-08-21
影响因子：
--
通讯作者：
Miller, Nicola
中科院分区：
文献类型：
--
作者：
Davoren, Pamela A.;McNeill, Roisin E.;Lowery, Aoife J.;Kerin, Michael J.;Miller, Nicola
microRNAs （miRNAs）的发现为已经复杂的调节基因表达的系统增加了额外的复杂性。这些单链RNA分子，长度为18-25个核苷酸，通过翻译抑制或mRNA切割负调控基因表达。特定mirna的异常表达与人类疾病有关的发现激发了人们对这些分子表达谱的兴趣。实时定量PCR （Real-time quantitative PCR, RQ-PCR）是一种灵敏、可重复的基因表达定量技术，目前已被用于分析细胞和组织中miRNA的表达。为了校正诸如起始模板数量、RNA质量和酶效率等系统变量，RQ-PCR数据通常归一化为内源性控制（EC）基因，理想情况下，该基因在整个测试样本集中稳定表达。适用于每种组织类型、治疗和疾病阶段的通用内源性对照尚未确定，也不太可能存在，因此，为了避免在表达数据的量化中引入进一步的错误，有必要在感兴趣的样本中验证候选ECs。虽然ECs已经在各种实验环境中被验证用于量化mRNA表达，但迄今为止还没有关于miRNA ECs在乳腺组织中表达谱验证的报道。在这项研究中，我们检测了5个miRNA基因（let-7a、miR-10b、miR-16、miR-21和miR-26b）和3个小核仁RNA基因（RNU19、RNU48和Z30）在恶性、良性和正常乳腺组织中的表达，以确定最合适的正常化策略。这是第一个通过RQ-PCR方法在人乳腺组织中鉴定可靠的ECs来分析miRNA的研究。
The discovery of microRNAs (miRNAs) added an extra level of intricacy to the already complex system regulating gene expression. These single-stranded RNA molecules, 18–25 nucleotides in length, negatively regulate gene expression through translational inhibition or mRNA cleavage. The discovery that aberrant expression of specific miRNAs contributes to human disease has fueled much interest in profiling the expression of these molecules. Real-time quantitative PCR (RQ-PCR) is a sensitive and reproducible gene expression quantitation technique which is now being used to profile miRNA expression in cells and tissues. To correct for systematic variables such as amount of starting template, RNA quality and enzymatic efficiencies, RQ-PCR data is commonly normalised to an endogenous control (EC) gene, which ideally, is stably-expressed across the test sample set. A universal endogenous control suitable for every tissue type, treatment and disease stage has not been identified and is unlikely to exist, so, to avoid introducing further error in the quantification of expression data it is necessary that candidate ECs be validated in the samples of interest. While ECs have been validated for quantification of mRNA expression in various experimental settings, to date there is no report of the validation of miRNA ECs for expression profiling in breast tissue. In this study, the expression of five miRNA genes (let-7a, miR-10b, miR-16, miR-21 and miR-26b) and three small nucleolar RNA genes (RNU19, RNU48 and Z30) was examined across malignant, benign and normal breast tissues to determine the most appropriate normalisation strategy. This is the first study to identify reliable ECs for analysis of miRNA by RQ-PCR in human breast tissue.
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影响因子：
14.9
作者：
Chen C;Ridzon DA;Broomer AJ;Zhou Z;Lee DH;Nguyen JT;Barbisin M;Xu NL;Mahuvakar VR;Andersen MR;Lao KQ;Livak KJ;Guegler KJ
通讯作者：
Guegler KJ
影响因子：
14.9
作者：
Griffiths-Jones, S
通讯作者：
Griffiths-Jones, S
影响因子：
5.3
作者：
Galardi, S;Fatica, A;Bozzoni, I
通讯作者：
Bozzoni, I
鉴定过氧蛋白6在乳腺癌进展中的功能作用。
DOI：
10.1186/bcr1789
发表时间：
2007
期刊：
Breast cancer research : BCR
影响因子：
--
作者：
Chang XZ;Li DQ;Hou YF;Wu J;Lu JS;Di GH;Jin W;Ou ZL;Shen ZZ;Shao ZM
通讯作者：
Shao ZM
影响因子：
14.9
作者：
Griffiths-Jones S;Grocock RJ;van Dongen S;Bateman A;Enright AJ
通讯作者：
Enright AJ