Housekeeping gene variability in normal and cancerous colorectal, pancreatic, esophageal, gastric and hepatic tissues
Housekeeping gene variability in normal and cancerous colorectal, pancreatic, esophageal, gastric and hepatic tissues
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DOI：
10.1016/j.mcp.2004.10.001
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发表时间：
2005-04-01
影响因子：
3.3
通讯作者：
Schilling, M
中科院分区：
文献类型：
--
作者：
Rubie, C;Kempf, K;Schilling, M
仔细的归一化对于准确定量活检组织样本中mRNA水平至关重要。通常，用内源性标准基因，主要是内源性内务基因（HKGs）来规范靶基因。然而，内源性内参基因的表达水平在不同组织和病理状态之间存在差异。因此，我们面临的挑战是确定一组内源性内参基因，其mRNA表达水平在正常组织和癌组织之间不会发生显著变化。采用实时荧光定量PCR （Quantitative real-time PCR, Q-RT-PCR）分析了21个经典管家基因在结直肠癌、胰腺癌、食管癌和胃癌以及肝转移癌中与正常组织相比的基因表达变异性。我们的研究结果表明，一些管家基因在一些研究组织中具有相对稳定的基因表达，但对于大多数被研究的HKGs，我们的数据揭示了不同组织和病理状态之间基因表达变异程度的明显差异。然而，对于所研究的五种组织中的每一种，我们都发现了一组以恒定水平表达的基因，从而代表了一组候选基因，我们可以推荐它们作为各自组织类型中的管家基因。综上所述，我们的研究结果可以为其他研究各种癌症的科学家提供指导，以进行组织特异性选择最佳管家基因（HKG），用于使靶基因表达正常化。(C) 2004 Elsevier Ltd.版权所有。
Careful normalization is essential for the accurate quantitation of mRNA levels in biopsy-sized tissue samples. Commonly, normalization of the target gene with an endogenous standard, mainly housekeeping genes (HKGs), is applied. However, differences in the expression levels of endogenous reference genes have been reported between different tissues and pathological states. Therefore, we were challenged to identify a set of endogenous reference genes whose mRNA expression levels would not change significantly between normal and cancerous tissues. Quantitative real-time PCR (Q-RT-PCR) analysis was applied to evaluate the variability in gene expression among 21 classical housekeeping genes in colorectal, pancreatic, esophageal and gastric cancer as well as in liver metastases in comparison to the corresponding normal tissue. Our results indicated that some housekeeping genes were candidates with relatively stable gene expression in several of the investigated tissues but for most of the HKGs under investigation our data have revealed distinct differences in the extent of variability in gene expression between the different tissues and pathological states. However, for each of the five tissues investigated we found a group Of genes that were expressed at a constant level thus representing a panel of candidates that we can recommend as housekeeping genes in the respective tissue types. In summary, our results can be used as guidance for other scientists studying various carcinomas for tissue-specific selection of the optimal housekeeping gene (HKG) to be used in normalizing target gene expression. (C) 2004 Elsevier Ltd. All rights reserved.