Identification of genes for normalization of quantitative real-time PCR data in ovarian tissues
Identification of genes for normalization of quantitative real-time PCR data in ovarian tissues
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DOI：
10.1093/abbs/gmq062
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发表时间：
2010-08-01
影响因子：
3.7
通讯作者：
Song, Haifeng
中科院分区：
文献类型：
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作者：
Fu, Jie;Bian, Lihong;Song, Haifeng
近年来，卵巢癌潜在的生物标志物和治疗靶点的确定越来越受到重视。然而，定量实时PCR的标准化对于获得准确的基因表达数据是重要的。我们研究了20个参考基因在不同条件下卵巢组织中的稳定性，以确定最适合这种应用的基因。本研究对52例正常、非恶性和原发性卵巢癌的卵巢组织样本中20个可能的参考基因的表达进行了表征。采用单因素方差分析（ANOVA）方法比较疾病进展所带来的候选基因变化。应用geNorm和Normands软件进一步验证了差异无统计学意义的基因的稳定性和适用性。结果表明：20个参比基因在不同卵巢组织中的表达量存在差异，而RPL4、RPLP0、HSPCB、TPT1、RPL13A、18S rRNA、PPIA、TBP和GUSB在不同卵巢组织中的表达量保持统计学稳定性。RPL4、RPLP0和HSPCB被证明是最稳定的参考基因，RPLP0和RPL4的组合应该被推荐为更可靠的标准化策略。
Increased attention has been paid to the determination of the potential biomarker and therapeutic target for ovarian cancer in recent years. However, the normalization of quantitative real-time PCR is important to obtain accurate gene expression data. We investigated the stability of 20 reference genes in ovarian tissues under different conditions to determine the most adequate for this application. The study characterized the expression of 20 possible reference genes among 52 ovarian tissue samples involving the normal, non-malignant, and primary ovarian carcinomas. One-way analysis of variance (ANOVA) method was used to compare the candidate gene changes brought about by the disease progression. The stability and suitability of the genes with no statistic difference were further validated employing geNorm and NormFinder softwares. Results showed that the expression levels of the 20 reference genes varied, while the RPL4, RPLP0, HSPCB, TPT1, RPL13A, 18S rRNA, PPIA, TBP, and GUSB kept statistic stability despite different ovarian tissue conditions. RPL4, RPLP0, and HSPCB were demonstrated as the most stable reference genes and the combination of the RPLP0 and RPL4 should be recommended as a much more reliable normalization strategy.