Identification of distinct gene expression profiles associated with treatment of LβT2 cells with gonadotropin-releasing hormone agonist using microarray analysis
Identification of distinct gene expression profiles associated with treatment of LβT2 cells with gonadotropin-releasing hormone agonist using microarray analysis
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DOI：
10.1016/s0378-1119(03)00446-3
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发表时间：
2003-04-10
期刊：
影响因子：
3.5
通讯作者：
Flynn, S
中科院分区：
文献类型：
--
作者：
Kakar, SS;Winters, SJ;Flynn, S
促性腺激素释放激素（GnRH）是一种在生殖过程中起关键作用的神经肽。近年来，人们已经清楚地认识到它也是一种抗扩散剂。GnRH类似物目前在临床上用于治疗前列腺癌以及子宫内膜异位症和性早熟。GnRH的靶细胞包括垂体前叶促性腺细胞和各种激素依赖性肿瘤细胞。然而，在这些细胞中仅鉴定出少数靶基因，并且对GnRH对它们的调节知之甚少。因此，我们在小鼠促性腺激素肿瘤细胞系（LbetaT2）中使用了定量微阵列分析来鉴定受GnRH调节的基因。用GnRH激动剂des-gly(10)[D-Ala(6)]GnRH （GnRHA）处理LbetaT2细胞1小时后，基因表达水平发生了1.3- 159倍的变化，与载体处理的细胞相比，共有232个基因表现出两倍或更大的表达变化。在这232个基因中，149个基因被上调，令人惊讶的是，83个基因被GnRHA下调。治疗24小时后，大多数在治疗1小时后表现出表达改变的基因的表达已恢复到基线水平。此外，在处理24小时后观察到不同的谱，有208个基因表现出两倍或更大的改变。其中95个上调，113个下调。在这项研究之前，大多数受影响的基因并不知道对GnRH有反应。研究发现，GnRHA治疗可影响多种基因的表达，包括致癌基因、编码转录因子的基因、离子通道蛋白、细胞骨架蛋白以及参与信号转导、细胞周期、细胞增殖和凋亡的其他蛋白质。通过微阵列分析发现的受GnRHA调控的6个基因的表达改变，通过半定量逆转录-聚合酶链反应证实。这是微阵列技术在GnRH调控基因全局图谱研究中的首次应用，并将被证明是未来分析GnRH调节促性腺激素表达和肿瘤细胞生长机制的有力工具。(C) 2003 Elsevier Science B.V.版权所有
Gonadotropin-releasing hormone (GnRH) is a neuropeptide that plays a pivotal role in reproductive processes. In recent years, it has become clear that it is also an anti-proliferative agent. GnRH analogs are now used clinically in the treatment of prostate cancer as well as endometriosis and precocious puberty. The target cells of GnRH include the gonadotropes of the anterior pituitary gland and the cells of various hormone-dependent tumors. Only a few target genes have been identified in these cells, however, and little is known concerning their regulation by GnRH. Therefore, we used a quantitative microarray assay to identify the genes that are regulated by GnRH in a murine gonadotrope tumor cell line (LbetaT2). Treatment of LbetaT2 cells with GnRH agonist des-gly(10)[D-Ala(6)]GnRH (GnRHA) for 1 h resulted in alterations in the levels of expression of genes that ranged in magnitude from 1.3- to 159-fold, with a total of 232 genes exhibiting a twofold or greater alteration in expression compared to vehicle treated cells. Of these 232 genes, 149 were up-regulated and, surprisingly, 83 were down-regulated by GnRHA treatment. After 24 h of treatment, the expression of most of the genes that had exhibited altered expression after 1 h of treatment had returned to baseline levels. Moreover, a different profile was observed after 24 h of treatment with 208 genes exhibiting a twofold or greater alteration. Of these, 95 were up-regulated and 113 down-regulated. Most of the affected genes were not known to be responsive to GnRH prior to this study. Treatment with GnRHA was found to affect the expression of a diverse range of genes, including oncogenes and those that encode transcription factors, ion channel proteins, and cytoskeletal proteins as well as other proteins that are involved in signal transduction, the cell cycle, cell proliferation and apoptosis. The altered expression of six of the genes that were found by microarray analysis to be regulated by GnRHA was confirmed by semiquantitative reverse transcriptase-polymerase chain reaction. This is first application of the microarray technique in the study of the global profile of genes regulated by GnRH, and should prove to be a powerful tool for future analysis of the mechanisms by which GnRH regulates the expression of gonadotropins and the growth of tumor cells. (C) 2003 Elsevier Science B.V. All rights reserved.