Identification of endogenous control genes for normalisation of real-time quantitative PCR data in colorectal cancer.
Identification of endogenous control genes for normalisation of real-time quantitative PCR data in colorectal cancer.
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DOI：
10.1186/1471-2199-11-12
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发表时间：
2010-02-01
影响因子：
--
通讯作者：
Miller N
中科院分区：
文献类型：
--
作者：
Kheirelseid EA;Chang KH;Newell J;Kerin MJ;Miller N
基因表达分析在肿瘤的诊断、预后和治疗护理中具有广泛的应用。相对定量是最广泛采用的方法，其中基因表达的定量相对于内源性表达控制（EC）基因规范化。基因表达可靠测定的核心是控制基因的选择。本研究的目的是评估一组候选EC基因，从中确定最稳定表达的基因，以规范来自原发性结直肠癌组织的RQ-PCR数据。分析了64例结直肠肿瘤和肿瘤相关正常标本中B2M、HPRT、GAPDH、ACTB、PPIA、HCRT、SLC25A23、DTX3、APOC4、RTDR1、KRTAP12-3、CHRNB4和MRPL19这13个候选EC基因的表达。选择CXCL12、FABP1、MUC2和PDCD4基因作为靶基因，比较各EC基因对基因表达的影响。采用描述性统计、geNorm、NormFinder和qBasePlus进行数据分析，发现候选EC基因之间存在显著差异。我们确定了最佳正常化需要两个基因，并确定B2M和PPIA是最稳定表达和最可靠的EC基因。本研究发现，两个EC基因（B2M和PPIA）的组合更准确地正常化了结直肠组织中的RQ-PCR数据。尽管这些控制基因在其他癌症研究中可能不是最佳的，但本文描述的方法可以作为识别其他癌症类型中有效ECs的模板。
Gene expression analysis has many applications in cancer diagnosis, prognosis and therapeutic care. Relative quantification is the most widely adopted approach whereby quantification of gene expression is normalised relative to an endogenously expressed control (EC) gene. Central to the reliable determination of gene expression is the choice of control gene. The purpose of this study was to evaluate a panel of candidate EC genes from which to identify the most stably expressed gene(s) to normalise RQ-PCR data derived from primary colorectal cancer tissue. The expression of thirteen candidate EC genes: B2M, HPRT, GAPDH, ACTB, PPIA, HCRT, SLC25A23, DTX3, APOC4, RTDR1, KRTAP12-3, CHRNB4 and MRPL19 were analysed in a cohort of 64 colorectal tumours and tumour associated normal specimens. CXCL12, FABP1, MUC2 and PDCD4 genes were chosen as target genes against which a comparison of the effect of each EC gene on gene expression could be determined. Data analysis using descriptive statistics, geNorm, NormFinder and qBasePlus indicated significant difference in variances between candidate EC genes. We determined that two genes were required for optimal normalisation and identified B2M and PPIA as the most stably expressed and reliable EC genes. This study identified that the combination of two EC genes (B2M and PPIA) more accurately normalised RQ-PCR data in colorectal tissue. Although these control genes might not be optimal for use in other cancer studies, the approach described herein could serve as a template for the identification of valid ECs in other cancer types.
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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：
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作者：
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影响因子：
2.9
作者：
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通讯作者：
Füzesi, L
影响因子：
10.3
作者：
O'Connell, JB;Maggard, MA;Ko, CY
通讯作者：
Ko, CY
影响因子：
3.5
作者：
Bustin, SA
通讯作者：
Bustin, SA