Androgens regulate SMAD ubiquitination regulatory factor-1 expression and prostate cancer cell invasion.
Androgens regulate SMAD ubiquitination regulatory factor-1 expression and prostate cancer cell invasion.
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DOI：
10.1002/pros.22935
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发表时间：
2015-05
期刊：
影响因子：
--
通讯作者：
Huang H
中科院分区：
文献类型：
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作者：
Gang X;Wang G;Huang H
前列腺癌(PCA)是美国最常见的男性癌症，是一种由激素驱动的疾病。雄激素已被认为是前列腺癌发生发展的主要促进剂。然而，雄激素在前列腺癌中的作用机制，特别是在前列腺癌细胞侵袭和转移中的作用机制尚不清楚。Smad泛素化调节因子-1(SMURF1)是一种C2-WW-Hect结构域的E3泛素连接酶，在癌细胞转移中发挥重要作用。雄激素与SMURF1表达之间是否存在联系尚不清楚。通过Western印迹分析和逆转录聚合酶链式反应(RT-PCR)检测雄激素对PCa细胞SMURF1表达的影响。通过电穿孔进行基因转染，以操纵所研究的蛋白质的表达水平。用染色质免疫沉淀(ChIP)法检测AR与SMURF1基因增强子的结合。创伤愈合实验和Matrigel侵袭实验分别检测细胞迁移和侵袭能力。我们发现，在前列腺癌细胞系中，雄激素可上调SMURF1的表达，并且雄激素的这种作用是通过雄激素受体(AR)介导的。我们进一步证明了雄激素在转录水平上调节SMURF1的表达，并提供了证据表明，AR通过与其含有典型的半雄激素反应元件(ARE)的增强子结合，在转录水平上激活SMURF1。最后，我们证明了SMURF1在雄激素诱导的PCa细胞侵袭中起重要作用。我们首次证明SMURF1是AR的一个真正的靶基因。我们的发现也提示SMURF1在前列腺癌转移中的潜在作用。
Prostate cancer (PCa) is the most commonly diagnosed male cancer in the United States and is a hormone-driven disease. Androgens have been recognized as a major promoter of PCa development and progression. However, the mechanism of androgen action in PCa, especially in PCa cell invasion and metastasis remains largely unclear. SMAD ubiquitination regulatory factor-1 (SMURF1) is a C2-WW-HECT-domain E3 ubiquitin ligase that plays important roles in cancer cell metastasis. Whether there is a relationship between androgens and SMURF1 expression is not known. The effect of androgens on the expression of SMURF1 in PCa cell lines was examined by Western blot analyses and reverse transcription-polymerase chain reaction (RT-PCR). Gene transfection was performed by electroporation to manipulate the expression levels of proteins studied. The binding of AR to the SMURF1 gene enhancer was determined by chromatin immunoprecipitation (ChIP) assay. Cell migration and invasion was measured by wound healing and Matrigel invasion assays, respectively. We found that expression of SMURF1 is upregulated by androgens in PCa cell lines and that this effect of androgens is mediated through the androgen receptor (AR). We further showed that androgens regulate SMURF1 expression at transcriptional level and provided evidence that AR transcriptionally activates SMURF1 by binding to its enhancer that contains a canonical half androgen responsive element (ARE). Finally, we demonstrated that SMURF1 is important for androgen-induced invasion of PCa cells. We demonstrate for the first time that SMURF1 is a bona fide target gene of the AR. Our findings also suggest a potential role of SMURF1 in PCa metastasis.