17beta-hydroxysteroid dehydrogenase type 1 modulates breast cancer protein profile and impacts cell migration
17beta-hydroxysteroid dehydrogenase type 1 modulates breast cancer protein profile and impacts cell migration
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DOI：
10.1186/bcr3207
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发表时间：
2012-01-01
影响因子：
7.4
通讯作者：
Lin, Sheng-Xiang
中科院分区：
文献类型：
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作者：
Aka, Juliette A.;Zerradi, Mouna;Lin, Sheng-Xiang
前言：人类17β-羟基类固醇脱氢酶1(17-β-HSD1)是一种类固醇转换酶，长期以来一直被认为在雌二醇合成和最近在二氢睾酮(DHT)失活中发挥关键作用，显示出促进乳腺癌细胞增殖的双重功能。此前，我们首次报道了该酶对内源性雌激素反应基因表达的影响。方法：将17β-HSD1稳定导入MCF7细胞，并与野生型MCF7细胞的蛋白质组进行比较。蛋白质组学研究采用双向凝胶电泳法，然后对差异表达的蛋白质斑点进行质谱分析。逆转录-实时定量聚合酶链式反应(RT-qPCR)检测单个基因的转录。17β-HSD1对MCF7细胞迁移的影响通过伤口愈合实验得到验证。结果：蛋白质组学数据表明，在17β-HSD1过表达后，超过59个蛋白质的表达受到调控。17β-HSD1调节与细胞生长控制相关的重要基因和蛋白的表达，如BRCA2和CDKN1A相互作用蛋白(BCCIP)和增殖细胞核抗原(PCNA)在MCF7-17βHSD1细胞中分别下调和上调。RT-qPCR结果显示，17β-HSD1使雌激素受体(ER)α和β的基因表达水平分别增加171%和120%，而使雄激素受体的基因表达水平降低%。有趣的是，17β-HSD1使肿瘤转移抑制基因nm23-H1的mRNA转录增加3.6倍，其蛋白表达与两种酶的表达密切相关。我们进一步证明了17β-HSD1的表达与MCF7细胞迁移的增加有关。结论：除了对重要基因的调控外，我们首次证明了17β-HSD1促进乳腺癌细胞迁移，尽管它对抗转移基因NM23有正向调节作用。这也与其刺激乳腺癌细胞生长有关，进一步证实了其对ER阳性乳腺癌的靶向性。这项研究中的新发现为未来关于17β-HSD1在乳腺癌进展和相关治疗中的作用的研究提出了几个方向。
Introduction: Human 17beta-hydroxysteroid dehydrogenase type 1 (17 beta-HSD1) is a steroid-converting enzyme that has long been known to play critical roles in estradiol synthesis and more recently in dihydrotestosterone (DHT) inactivation, showing a dual function that promotes breast cancer cell proliferation. Previously, we reported the first observation of the influence of the enzyme on endogenous estrogen-responsive gene expression. Here, we demonstrate the impact of 17 beta-HSD1 expression on the breast cancer cell proteome and investigate its role in cell migration.Methods: 17 beta-HSD1 was stably transfected in MCF7 cells and the proteome of the generated cells overexpressing 17 beta-HSD1 (MCF7-17 beta HSD1 cells) was compared to that of the wild type MCF7 cells. Proteomics study was performed using two-dimensional gel electrophoresis followed by mass spectrometry analysis of differentially expressed protein spots. Reverse transcription quantitative real-time PCR (RT-qPCR) was used to investigate the transcription of individual gene. The effect of 17 beta-HSD1 on MCF7 cell migration was verified by a wound-healing assay.Results: Proteomic data demonstrate that the expression of more than 59 proteins is modulated following 17 beta-HSD1 overexpression. 17 beta-HSD1 regulates the expression of important genes and proteins that are relevant to cell growth control, such as BRCA2 and CDKN1A interacting protein (BCCIP) and proliferating cell nuclear antigen (PCNA) which are down-and upregulated in MCF7-17 beta HSD1 cells, respectively. RT-qPCR data reveal that 17 beta-HSD1 increases the mRNA levels of estrogen receptors (ER) alpha and beta by 171 and 120%, respectively, while decreasing that of the androgen receptor by 64%. Interestingly, 17 beta-HSD1 increases the mRNA transcript (by 3.6 times) and the protein expression of the metastasis suppressor gene nm23-H1 and the expression of the two enzymes are closely correlated. We have further shown that 17 beta-HSD1 expression is associated with an increase of MCF7 cell migration.Conclusions: In addition to the regulation of important genes, we have demonstrated for the first time that 17 beta-HSD1 increases breast cancer cell migration, in spite of its positive regulation of the antimetastatic gene NM23. This is also correlated to its stimulation of breast cancer cell growth, further confirming its targeting in ER positive breast cancer. The novel findings in this study suggest several directions for future research on the contribution of 17 beta-HSD1 to breast cancer progression and related treatment.