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通过竞争性聚合酶链反应扩增评估人在位和异位子宫内膜基质细胞中碱性成纤维细胞生长因子信使核糖核酸水平。

Basic fibroblast growth factor messenger ribonucleic acid levels in eutopic and ectopic human endometrial stromal cells as assessed by competitive polymerase chain reaction amplification.

作者信息

Di Blasio A M, Centinaio G, Carniti C, Somigliana E, Viganò P, Vignali M

机构信息

Laboratorio di Biologia Molecolare, Centro Auxologico Italiano, Milano, Italy.

出版信息

Mol Cell Endocrinol. 1995 Dec 29;115(2):169-75. doi: 10.1016/0303-7207(95)03687-3.

Abstract

Previous studies have localized basic fibroblast growth factor (bFGF) and its mRNA in normal and neoplastic endometrium but the expression of bGFG mRNA in endometriosis cells is virtually unknown. Our purpose was to investigate the presence of bFGF and its receptor mRNAs in highly purified primary cultures of stromal cells isolated from six eutopic endometrial samples obtained from patients without evidence of endometriosis and five endometriosis cyst linings. Using reverse transcriptase-polymerase chain reaction (RT-PCR), single major DNA bands of the expected sizes for bFGF and its receptor (354 and 661 bp, respectively) were detected in both endometrial and endometriosis samples. A competitive RT-PCR, that uses coprimer extension and amplification of a bovine RNA as an internal standard, was developed for semiquantitative estimation of bFGF gene expression. The target to standard mean ratios +/- SEM in six normal endometrial samples and in five endometriosis cultures were 26.7 +/- 10.7 and 9.2 +/- 3.0, respectively. Furthermore, when data were analyzed according to the cyst diameter, levels of bFGF mRNA resulted statistically higher (P < 0.05) in bigger cysts when compared to those detected in smaller ones (16 +/- 2.7 and 4.7 +/- 1.8, respectively). These results demonstrate that the genes coding for bFGF and its receptor are expressed in endometriosis cells, but levels of bFGF mRNA are generally lower than those detected in their eutopic counterpart. Moreover, they indicate that endometriosis cells derived from large cysts have increased bFGF mRNA levels. Thus, bFGF could be one of the factors responsible for a more or less active behavior of the endometnotic lesion.

摘要

以往的研究已将碱性成纤维细胞生长因子(bFGF)及其mRNA定位在正常和肿瘤性子宫内膜中,但bGFG mRNA在子宫内膜异位症细胞中的表达实际上尚不清楚。我们的目的是研究从6例无子宫内膜异位症证据的患者获得的在位子宫内膜样本和5例子宫内膜异位症囊肿内膜中分离出的高度纯化的基质细胞原代培养物中bFGF及其受体mRNA的存在情况。使用逆转录聚合酶链反应(RT-PCR),在子宫内膜和子宫内膜异位症样本中均检测到了预期大小的bFGF及其受体的单一主要DNA条带(分别为354和661 bp)。开发了一种竞争性RT-PCR,该方法使用共引物延伸和扩增牛RNA作为内标,用于bFGF基因表达的半定量估计。6例正常子宫内膜样本和5例子宫内膜异位症培养物中的靶标与标准平均比值±SEM分别为26.7±10.7和9.2±3.0。此外,当根据囊肿直径分析数据时,与较小囊肿中检测到的水平相比,较大囊肿中bFGF mRNA的水平在统计学上更高(P<0.05)(分别为16±2.7和4.7±1.8)。这些结果表明,编码bFGF及其受体的基因在子宫内膜异位症细胞中表达,但bFGF mRNA的水平通常低于在位对应物中检测到的水平。此外,它们表明来自大囊肿的子宫内膜异位症细胞的bFGF mRNA水平升高。因此,bFGF可能是导致子宫内膜异位症病变或多或少活跃行为的因素之一。

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