Resistin induces breast cancer cells epithelial to mesenchymal transition (EMT) and sternness through both adenylyl cyclase-associated protein 1 (CAP1)-dependent and CAP1-independent mechanisms
Resistin induces breast cancer cells epithelial to mesenchymal transition (EMT) and sternness through both adenylyl cyclase-associated protein 1 (CAP1)-dependent and CAP1-independent mechanisms
复制标题
DOI：
10.1016/j.cyto.2019.04.016
复制
发表时间：
2019-08-01
期刊：
影响因子：
3.8
通讯作者：
Poretsky, Leonid
中科院分区：
文献类型：
--
作者：
Avtanski, Dimiter;Garcia, Anabel;Poretsky, Leonid
众所周知，绝经后女性乳腺癌的发病率和转移与肥胖有关，但这种关联背后的分子机制尚不清楚。我们在体外研究了脂肪因子抵抗素对乳腺癌细胞上皮间质转化（EMT）和干性的影响。先前的报告表明，抵抗素的炎症作用是由腺苷酸环化酶相关蛋白 1 (CAP1) 介导的，CAP1 是其受体。作为我们研究的模型，我们使用 MCF-7 和 MDA-MB-231 乳腺癌以及 MCF-10A 乳腺上皮细胞。我们发现，在 MCF-7 细胞中，抵抗素可增加 MCF-7 和 MDA-MB-231 细胞的迁移，并通过 F-肌动蛋白丝的重组诱导细胞突起的形成。抵抗素上调参与 EMT 的间质标志物（SNAIL、SLUG、ZEB1、TWIST1、纤连蛋白和波形蛋白）的表达，并下调上皮标志物（E-钙粘蛋白和claudin-1）的表达。抵抗素还增强了 SNAIL 蛋白的核转位，表明 EMT 重编程的启动。我们进一步在非致癌乳腺上皮 MCF-10A 细胞中诱导 EMT，证明抵抗素对 EMT 的影响不是乳腺癌细胞特异性的。为了评估抵抗素诱导的EMT是否依赖于CAP1，我们使用siRNA方法沉默MCF-7细胞中的CAP1基因。结果表明，当 CAP1 被沉默时，抵抗素诱导的 SNAIL、ZEB1 和波形蛋白表达以及 SNAIL 和 ZEB1 核转位均被消除。此外，CAP1 沉默导致 MCF-7 细胞迁移受到抑制。我们对与癌症干细胞 (CSC)、多能性和转移相关的 84 个基因的表达进行了定量 PCR 阵列分析，并选择了一组受抵抗素调节的基因（ALDH1A1、ITGA4、LIN28B、SMO、KLF17、PTPRC、PROM1、SIRT1 和 PECAM1）。进一步的实验表明，抵抗素对其中一些基因（PROM1、PTPRC、KLF17、SIRT1 和 PECAM1）表达的影响也依赖于 CAP1。我们的结果表明，抵抗素通过诱导 EMT 和干性来促进乳腺癌细胞的转移潜力，其中一些效应是由 CAP1 介导的。
Breast cancer incidence and metastasis in postmenopausal women are known to associate with obesity, but the molecular mechanisms behind this association are largely unknown. We investigated the effect of adipokine resistin on epithelial to mesenchymal transition (EMT) and stemness in breast cancer cells in vitro. Previous reports demonstrated that the inflammatory actions of resistin are mediated by the adenylyl cyclase-associated protein 1 (CAP1), which serves as its receptor. As a model for our study, we used MCF-7 and MDA-MB-231 breast cancer and MCF-10A breast epithelial cells. We showed that in MCF-7 cells resistin increases the migration of MCF-7 and MDA-MB-231 cells and induces the formation of cellular protrusions through reorganization of F-actin filaments. Resistin upregulated the expression of mesenchymal markers involved in EMT (SNAIL, SLUG, ZEB1, TWIST1, fibronectin, and vimentin), and downregulated those of epithelial markers (E-cadherin and claudin-1). Resistin also potentiated the nuclear translocation of SNAIL protein, indicating initiation of EMT reprogramming. We further induced EMT in non-carcinogenic breast epithelial MCF-10A cells demonstrating that the effects of resistin on EMT were not breast cancer cell specific. In order to assess whether resistin-induced EMT depends on CAP1, we used siRNA approach to silence CAP1 gene in MCF-7 cells. Results demonstrated that when CAP1 was silenced, the induction of SNAIL, ZEB1 and vimentin expression by resistin as well as SNAIL and ZEB1 nuclear translocation, were abolished. Additionally, CAP1 silencing resulted in a suppression of MCF-7 cells migration. We performed quantitative PCR array profiling the expression of 84 genes related to cancer stem cells (CSC), pluripotency and metastasis and selected a set of genes (ALDH1A1, ITGA4, LIN28B, SMO, KLF17, PTPRC, PROM1, SIRT1, and PECAM1) that were modulated by resistin. Further experiments demonstrated that the effect of resistin on the expression of some of these genes (PROM1, PTPRC, KLF17, SIRT1, and PECAM1) was also dependent on CAP1. Our results demonstrate that resistin promotes the metastatic potential of breast cancer cells by inducing EMT and stemness and some of these effects are mediated by CAP1.