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Evaluation of Combined Quantification of PCA3 and AMACR Gene Expression for Molecular Diagnosis of Prostate Cancer in Moroccan Patients by RT-qPCR

机译:RT-qPCR联合定量分析PCA3和AMACR基因表达对摩洛哥前列腺癌分子诊断的价值

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摘要

Prostate cancer (PCa) remains one of the most widespread and perplexing of all human malignancies. Assessment of gene expression is thought to have an important impact on cancer diagnosis, prognosis and therapeutic decisions. In this context, we explored combined expression of PCa related target genes AMACR and PCA3 in 126 formalin fixed paraffin embedded prostate tissues (FFPE) from Moroccan patients, using quantitative real time reverse transcription-PCR (RT-qPCR). This quantification required data normalization accomplished using stably expressed reference genes (RGs). A panel of twelve RG was assessed, data being analyzed using GenEx V6 based on geNorm, NormFinder and statistical methods. Accordingly, the hnRNP A1 gene was identified and selected as the most stably expressed RG for reliable and accurate gene expression quantification in prostate tissues. The ratios of both PCA3 and AMACR gene expression relative to that of the hnRNP A1 gene were calculated and the performance of each target gene for PCa diagnosis was evaluated using receiver-operating characteristics. PCA3 and AMACR mRNA quantification based on RT-qPCR may prove useful in PCa diagnosis. Of particular interesting, combining PCA3 and AMACR quantification improved PCa prediction by increasing sensitivity with retention of good specificity.
机译:前列腺癌(PCa)仍然是所有人类恶性肿瘤中分布最广泛,最令人困惑的问题之一。基因表达的评估被认为对癌症的诊断,预后和治疗决策具有重要影响。在这种情况下,我们使用定量实时逆转录PCR(RT-qPCR)探索了摩洛哥患者126个福尔马林固定石蜡包埋前列腺组织(FFPE)中PCa相关靶基因AMACR和PCA3的联合表达。此量化需要使用稳定表达的参考基因(RGs)实现的数据标准化。评估了一组十二个RG,使用基于geNorm,NormFinder和统计方法的GenEx V6对数据进行了分析。因此,hnRNP A1基因被鉴定并选择为最稳定表达的RG,用于前列腺组织中可靠,准确的基因表达定量。计算了PCA3和AMACR基因相对于hnRNP A1基因的表达比率,并使用接收者操作特征评估了每个目标基因诊断PCa的性能。基于RT-qPCR的PCA3和AMACR mRNA定量可证明对PCa诊断有用。尤其有趣的是,结合PCA3和AMACR定量可提高敏感性并保留良好的特异性,从而改善PCa预测。

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