Aromatase gene expression and its exon I usage in human breast tumors. Detection of aromatase messenger RNA by reverse transcription-polymerase chain reaction.
Aromatase gene expression and its exon I usage in human breast tumors. Detection of aromatase messenger RNA by reverse transcription-polymerase chain reaction.
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芳香酶基因表达及其外显子 I 在人类乳腺肿瘤中的使用。
DOI：
10.1016/s0960-0760(96)00100-8
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发表时间：
1996
期刊：
影响因子：
--
通讯作者：
Chen,S
中科院分区：
文献类型：
--
作者：
Zhou,C;Zhou,D;Esteban,J;Murai,J;Siiteri,PK;Wilczynski,S;Chen,S
应用逆转录-聚合酶链反应（RT-PCR）方法，对70例乳腺癌组织中芳香化酶的表达进行了研究。RT-PCR分析，使用两个寡核苷酸引物来自人类芳香化酶基因的外显子II显示，芳香化酶mRNA的检测，但三个组织标本。此外，引物定向RT-PCR进行，以确定在这些乳腺肿瘤标本芳香化酶mRNA外显子I的使用。分析显示，外显子I.3和PII是存在于从乳腺肿瘤分离的芳香酶mRNA中的两个主要外显子I，这表明启动子I.3和II是驱动乳腺癌和周围脂肪基质细胞中芳香酶表达的主要启动子。RT-PCR分析还检测到两种产物，I.3A（长度为334 bp）和I.3B（长度为222 bp），当使用来自外显子I.3的引物和来自外显子II的反向引物进行时。这些产物的核苷酸序列已经确定，并表明I.3A含有一个以前被认为是内含子的区域。此外，从8对乳腺肿瘤和邻近正常组织标本中分离的RNA进行RT-PCR分析，以评估外显子I的使用和I. 3A-和I. 3B-含有芳香化酶RNA信息在乳腺肿瘤和邻近正常组织中的分布。结果表明，I.3B和I.3A的信息主要存在于乳腺肿瘤和邻近的正常组织，分别。最后，通过引物指导的RT-PCR分析检查了八种细胞系（皮肤成纤维细胞、MCF-7、MDA-MB-231、T-47 D、SK-BR-3、SKF、OVCAR-3和人脂肪基质细胞）中芳香酶表达的外显子I/启动子使用。这些研究为进一步评价乳腺肿瘤芳香化酶表达和雌激素生物合成的调控机制提供了基础。
The expression of aromatase in human breast tumors has been studied by the reverse-transcription polymerase chain reaction (RT-PCR) method on 70 breast tissue specimens. An RT-PCR analysis using two oligonucleotide primers derived from the exon II of the human aromatase gene revealed that aromatase mRNA was detected in all but three tissue specimens. Furthermore, primer-directed RT-PCR was performed to determine the exon I usage in aromatase mRNA in these breast tumor specimens. The analysis has revealed that exons I.3 and PII are the two major exon Is present in aromatase mRNA isolated from breast tumors, suggesting that promoters I.3 and II are the major promoters driving aromatase expression in breast cancer and surrounding adipose stromal cells. The RT-PCR analysis also detected two products, I.3A (334 bp in length) and I.3B (222 bp in length), when it was carried out using a primer derived from exon I.3 and a reverse primer derived from exon II. The nucleotide sequences of these products have been determined and indicate that I.3A contains a region which was previously thought to be an intron. In addition, RT-PCR analyses of RNA isolated from eight pairs of breast tumor and neighboring normal tissue specimens were performed to evaluate the exon I usage and the distribution of I.3A- and I.3B-containing aromatase RNA messages in breast tumor and neighboring normal tissues. The results suggest that I.3B- and I.3A-containing messages are mainly present in breast tumor and neighboring normal tissues, respectively. Finally, the exon I/promoter usage for aromatase expression in eight cell lines (skin fibroblast, MCF-7, MDA-MB-231, T-47D, SK-BR-3, JAR, OVCAR-3, and human adipose stromal cells) was examined by primer-directed RT-PCR analyses. These studies provide a basis for further evaluation of the control mechanism of aromatase expression and estrogen biosynthesis in breast tumors.