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High-throughput differential screening of mRNAs by serial analysis of gene expression: decreased expression of trefoil factor 3 mRNA in thyroid follicular carcinomas

机译:通过基因表达系列分析的高通量差异筛选mRNA:甲状腺滤泡癌中三叶因子3 mRNA的表达降低

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To find mRNAs whose expression differs between thyroid follicular adenomas and carcinomas, a high-throughput analysis of mRNAs in these two tumours was performed. This method, named high-throughput differential screening by serial analysis of gene expression (HDSS), combines a modified method of serial analysis of gene expression (SAGE) and real-time quantitative reverse transcription polymerase chain reaction (RT–PCR). A total of 40 candidate tag sequences that showed extremely different expression levels between a follicular carcinoma and a follicular adenoma in the SAGE analysis were analysed by real-time quantitative RT–PCR, using RNAs from an additional four typical follicular carcinomas and adenomas. One sequence tag that represents trefoil factor 3 (TFF3) mRNA showed a clear difference in its expression level between adenomas and carcinomas. The expression levels of TFF3 mRNA in 48 follicular adenomas and 29 follicular carcinomas were measured by real-time quantitative RT–PCR using a specific probe for TFF3. They were significantly decreased in follicular carcinomas, especially in widely invasive types and those with evident metastases. These results indicate that the decreased expression of TFF3 mRNA is a marker of follicular carcinomas, especially those with a high risk of invasion or metastasis.
机译:为了发现在甲状腺滤泡性腺瘤和癌之间表达不同的mRNA,对这两个肿瘤中的mRNA进行了高通量分析。这种方法被称为通过基因表达序列分析(HDSS)进行的高通量差异筛选,结合了基因表达序列分析(SAGE)和实时定量逆转录聚合酶链反应(RT-PCR)的改良方法。通过实时定量RT-PCR,使用来自另外四种典型滤泡癌和腺瘤的RNA,分析了总共40个在SAGE分析中显示出滤泡癌和滤泡腺瘤之间表达水平完全不同的候选标签序列。代表三叶因子3(TFF3)mRNA的一个序列标签在腺瘤和癌之间显示出明显的表达水平差异。通过实时定量RT-PCR,使用TFF3的特异性探针,测量了48个滤泡性腺瘤和29个滤泡性癌中TFF3 mRNA的表达水平。在滤泡癌中,尤其是在广泛浸润型和有明显转移的滤泡癌中,它们明显减少。这些结果表明,TFF3 mRNA表达的降低是滤泡癌的标志,特别是那些具有高浸润或转移风险的滤泡癌。

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