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运用逆转录-聚合酶链反应检测乳腺肿瘤中芳香化酶和角质形成细胞生长因子的表达

Detection of aromatase and keratinocyte growth factor expression in breast tumors using reverse transcription-polymerase chain reaction.

作者信息

Koos R D, Banks P K, Inkster S E, Yue W, Brodie A M

机构信息

Department of Physiology, School of Medicine, University of Maryland, Baltimore 21201.

出版信息

J Steroid Biochem Mol Biol. 1993 Apr;45(4):217-25. doi: 10.1016/0960-0760(93)90335-t.

Abstract

It has been proposed that local production of estrogen may contribute to breast tumor growth, in part by regulating growth factor production. In view of this, we studied the expression of mRNAs for aromatase cytochrome P-450, the enzyme which catalyzes estrogen synthesis, and keratinocyte growth factor (KGF), a heparin-binding growth factor specific for epithelial cells, in breast tumors. In order to detect mRNAs of low-abundance, reverse transcription-polymerase chain reaction (RT-PCR) was used. RNA from breast tumors and normal breast tissue was reverse transcribed and then amplified using oligonucleotide primers for human aromatase or KGF. Twelve of 15 breast tumor samples yielded variable amounts of aromatase PCR product, but consistently strong KGF PCR signals. Of the three aromatase mRNA-negative samples, two gave weak KGF signals while one was negative for KGF. Both aromatase and KGF transcripts were also detected in all five normal breast tissue samples examined. These results indicate that a high proportion of breast tumors have the potential to produce aromatase and KGF, both of which could play important roles in their growth. The results also suggest that RT-PCR can be used to evaluate local expression of growth mediators in tumors.

摘要

有人提出,局部雌激素的产生可能在一定程度上通过调节生长因子的产生来促进乳腺肿瘤生长。鉴于此,我们研究了乳腺肿瘤中芳香化酶细胞色素P - 450(催化雌激素合成的酶)和角质形成细胞生长因子(KGF,一种对上皮细胞特异的肝素结合生长因子)的mRNA表达。为了检测低丰度mRNA,采用了逆转录 - 聚合酶链反应(RT - PCR)。来自乳腺肿瘤和正常乳腺组织的RNA进行逆转录,然后使用人芳香化酶或KGF的寡核苷酸引物进行扩增。15个乳腺肿瘤样本中有12个产生了不同量的芳香化酶PCR产物,但KGF PCR信号始终很强。在3个芳香化酶mRNA阴性样本中,2个给出较弱的KGF信号,而1个KGF为阴性。在所检测的所有5个正常乳腺组织样本中也检测到了芳香化酶和KGF转录本。这些结果表明,高比例的乳腺肿瘤有产生芳香化酶和KGF的潜力,这两者在其生长中可能起重要作用。结果还表明,RT - PCR可用于评估肿瘤中生长介质的局部表达。

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