Bergapten induces ER depletion in breast cancer cells through SMAD4-mediated ubiquitination.
Bergapten induces ER depletion in breast cancer cells through SMAD4-mediated ubiquitination.
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Bergapten通过SMAD4介导的泛素化诱导乳腺癌细胞的ER耗竭。
DOI：
10.1007/s10549-012-2282-3
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发表时间：
2012-11
影响因子：
3.8
通讯作者：
Ando, S.
中科院分区：
文献类型：
--
作者：
Panno, M. L.;Giordano, F.;Rizza, P.;Pellegrino, M.;Zito, D.;Giordano, C.;Mauro, L.;Catalano, S.;Aquila, S.;Sisci, D.;De Amicis, F.;Vivacqua, A.;Fuqua, S. W. A.;Ando, S.
ER-α功能对于正常乳腺的发育以及乳腺癌细胞的发展过程都是至关重要的。靶向受体水平有助于调节细胞内雌激素效应的信号。在内质网α和转化生长因子-β下游分子Smad2、Smad3和Smad4之间存在复杂的交叉调节，它们可以结合ERα并调节其信号转导。因此，识别能够影响后一种效果的天然抗癌药物可能为乳腺癌的治疗提供替代选择。考虑到我们以前发表的数据，我们希望研究5-甲氧补骨脂素(贝加平)对内质网α和转化生长因子-β通路的影响。采用逆转录聚合酶链式反应和免疫印迹法检测灯盏花素对雌激素受体α表达蛋白和转化生长因子-β下游信号分子的影响。Smad4和转化生长因子-βRII的siRNA也被用来评估它们在贝加普通诱导的反应中的作用。我们报道了一种含有香豆素的化合物Bergapten能有效地耗尽MCF-7乳腺癌敏感细胞和三苯氧胺耐药克隆中的ERα。贝加平治疗后ERα蛋白的减少是泛素-蛋白酶体途径的结果，MG-132的使用证明了这一点。与ER抗体的IP实验表明，该蛋白与Smad4和多泛素具有物理相互作用，并且在bergapten作用下，连接Smad4的泛素化受体的数量更多。在这个过程中，Smad4发挥了关键的作用，因为观察到在乳腺癌细胞中，Smad4的沉默导致内源性ERα在对照和bergapten处理的细胞中的表达增加，与野生型细胞相比。在siRNA转化生长因子-βRII细胞中也证实了同样的结果。结果提示转化生长因子α/Smad4对乳腺癌细胞ERβ的负性调节作用，提示Smad4蛋白参与了佛波酯诱导的ERα的降解。我们认为贝加普腾可能是一种有效的天然抗肿瘤药物，能够耗尽乳腺癌他莫昔芬敏感和耐药细胞中的ERα，从而重新训练针对ERα的膜信号的作用，从而使其有丝分裂潜能。
ERα function is crucial for development of normal mammary gland as well as in the process of progression of breast cancer cells. Signals that target receptor levels contribute to regulate estrogens effects in the cells. An intricate cross-regulation has been documented between ERα and TGF-β down-stream molecules: SMAD2, SMAD3 and SMAD4, that can bind ERα and regulate their signaling. Thus, identification of natural anticancer drugs able to influence the latter effect might provide alternative choices for breast cancer treatment. Taking into account our previous published data we wanted to study the effect of 5-Methoxypsoralen (bergapten) on ERα and on TGF-β pathway. RT-PCR and W.B. were performed to evaluate the effect of bergapten on the ERα expression protein and the TGF-β –down stream signaling molecules. siRNA for Smad4 and TGF- β RII was also done to evaluate their involvement on the bergapten-induced responses. We reported that bergapten, a coumarin containing compound, effectively depletes ERα in MCF-7 breast cancer sensitive cells and in tamoxifen-resistant clone. The decrease of ERα protein after bergapten treatment results from the ubiquitine-proteasome pathway as demonstrated by the use of MG-132. IP experiments with ER antibody, demonstrated that the protein has physical interaction with SMAD4 and poly-ubiquitine and the amount of ubiquitinated receptor, linked to SMAD4, is greater under bergapten. The crucial role played by SMAD4, in this process, emerges from the observation that in breast cancer cells, silencing of SMAD4, resulted in increased expression of endogenous ERα in both control and bergapten-treated cells, compared to wild type cells. The same results were confirmed in siRNA TGF-β RII cells. The results suggest a novel negative regulation of ERα by TGF-β/SMAD4 in breast cancer cells and indicate that the SMAD4 protein is involved in the degradation of ERα induced by bergapten. We propose that bergapten may efficiently act as a natural antitumoral agent, able to deplete ERα from breast cancer tamoxifen-sensitive and resistant cells, thereby retraining the effect of membrane signals targeting ERα and in such way its mitogenic potentiality.
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