Identification of suitable reference genes for gene expression measurement in uterine sarcoma and carcinosarcoma tumors
Identification of suitable reference genes for gene expression measurement in uterine sarcoma and carcinosarcoma tumors
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DOI：
10.1016/j.clinbiochem.2012.01.001
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发表时间：
2012-03-01
影响因子：
2.8
通讯作者：
Bidzinski, Mariusz
中科院分区：
文献类型：
--
作者：
Kowalewska, Magdalena;Danska-Bidzinska, Anna;Bidzinski, Mariusz
目的：迄今为止，尚未在子宫体的癌细胞瘤和非上皮恶性肿瘤中鉴定出用于标准化实时RT-PCR（qRT-PCR）测定的合适的参考基因。本研究的目的是选择合适的参考文献中的基因表达研究在这些tumors.Materials和方法：我们使用的RNA提取物从75个组织样本，代表50个肿瘤和25个片段的正常子宫组织从50例混合瘤，平滑肌肉瘤和子宫间质肉瘤治疗。对5个潜在参考（管家）基因（即B2 M、HMBS、HPRT 1、TBP和UBC）进行qRT-PCR。结果：用geNorm软件对基因表达数据进行分析，发现HPRT 1是所有组织中最稳定的参考基因，其次是UBC和HMBS。当按疾病分层时，结果仍然指向HPRT 1作为在混合瘤以及平滑肌和间质肉瘤中保留最大稳健性的基因。结论：我们的工作是第一份关于qRT-PCR在混合瘤、平滑肌肉瘤和子宫间质肉瘤中应用的参考基因选择的报告。提供了三种类型肿瘤的候选基因的稳定性值的排名，并可能作为这些罕见实体的未来基因表达研究的有价值的指导。(C)2012年加拿大临床化学家协会。由爱思唯尔公司出版All rights reserved.
Objective: To date no suitable reference genes have been identified in carcinosarcomas and non-epithelial malignant tumors of corpus uteri for normalizing real-time RT-PCR (qRT-PCR) assays. The purpose of this study was to select appropriate references for gene expression studies in these tumors.Materials and methods: We used RNA extracts from 75 tissue samples, representing 50 tumors and 25 fragments of normal uterine tissues obtained from 50 patients treated for mixed tumors, smooth muscle sarcoma and stromal sarcoma of the uterus. qRT-PCR for five potential reference (housekeeping) genes, namely B2M, HMBS, HPRT1, TBP and UBC, was performed. The expression stability of these genes was assayed using geNorm software application.Results: The analysis of gene expression data with geNorm identified HPRT1 as the most stable reference gene, followed by UBC and HMBS, for all the investigated tissues. When stratified by disease, the results still pointed at HPRT1 as the gene that retained the greatest robustness in mixed tumors as well as in smooth muscle and stromal sarcomas.Conclusions: Our work is the first report on reference gene selection for qRT-PCR applications in mixed tumors, smooth muscle sarcoma and stromal sarcoma of the uterus. A ranking of candidate genes' stability values for the three types of tumors is provided and might serve as a valuable guide for future gene expression studies of these rare entities. (C) 2012 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.