Evaluation of lumican effects on morphology of invading breast cancer cells, expression of integrins and downstream signaling
Evaluation of lumican effects on morphology of invading breast cancer cells, expression of integrins and downstream signaling
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DOI：
10.1111/febs.15289
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发表时间：
2020-03-31
期刊：
影响因子：
5.4
通讯作者：
Brezillon, Stephane
中科院分区：
文献类型：
--
作者：
Karamanou, Konstantina;Franchi, Marco;Brezillon, Stephane
小的富含亮氨酸的蛋白聚糖光蛋白聚糖调节乳腺癌细胞的雌激素受体（ER）相关的功能特性，基质大分子的表达和上皮间质转化。然而，目前尚不清楚ER依赖性Lumican对乳腺癌细胞的作用是否与整合素及其细胞内信号通路的表达有关。在这里，我们分析了lumican在三种乳腺癌细胞系中的作用：高转移性ER β阳性MDA-MB-231，具有各自ER β抑制的细胞（shER β MDA-MB-231）和低侵袭性ER α阳性MCF-7/c乳腺癌细胞。进行扫描电子显微镜、共聚焦显微镜、实时PCR、蛋白质印迹和细胞粘附测定。还在三维胶原培养物中研究了Lumican对乳腺癌细胞形态的影响。Lumican处理诱导细胞-细胞接触和细胞分组，并抑制微泡和微绒毛的形成。还研究了细胞表面粘附受体CD 44、其同种型和变体、透明质酸（HA）和HA降解酶的表达。Lumican抑制CD 44和HA结合酶的表达，其对细胞粘附的作用揭示了整合素α 1、α 2、α 3、α V β 3和α V β 5在MDA-MB-231细胞中的主要作用，但在MCF-7/c细胞中不起作用。与MCF-7/c细胞相比，Lumican上调了MDA-MB-231和shER β MDA-MB-231中α 2和β 1整联蛋白亚基的表达。发现整合素的下游信号传导途径如FAK、ERK 1/2 MAPK 42/44和Akt被Lumican下调。我们的数据揭示了Lumican在浸润性乳腺癌中抗癌活性的分子机制。
The small leucine-rich proteoglycan lumican regulates estrogen receptors (ERs)-associated functional properties of breast cancer cells, expression of matrix macromolecules, and epithelial-to-mesenchymal transition. However, it is not known whether the ER-dependent lumican effects on breast cancer cells are related to the expression of integrins and their intracellular signaling pathways. Here, we analyzed the effects of lumican in three breast cancer cell lines: the highly metastatic ER beta-positive MDA-MB-231, cells with the respective ER beta-suppressed (shER beta MDA-MB-231), and lowly invasive ER alpha-positive MCF-7/c breast cancer cells. Scanning electron microscopy, confocal microscopy, real-time PCR, western blot, and cell adhesion assays were performed. Lumican effects on breast cancer cell morphology were also investigated in 3-dimensional collagen cultures. Lumican treatment induced cell-cell contacts and cell grouping and inhibited microvesicles and microvilli formation. The expression of the cell surface adhesion receptor CD44, its isoform and variants, hyaluronan (HA), and HA synthases was also investigated. Lumican inhibited the expression of CD44 and HA synthases, and its effect on cell adhesion revealed a major role of alpha 1, alpha 2, alpha 3, alpha V beta 3, and alpha V beta 5 integrins in MDA-MB-231 cells, but not in MCF-7/c cells. Lumican upregulated the expression of alpha 2 and beta 1 integrin subunits both in MDA-MB-231 and in shER beta MDA-MB-231 as compared to MCF-7/c cells. Downstream signaling pathways for integrins, such as FAK, ERK 1/2 MAPK 42/44, and Akt, were found to be downregulated by lumican. Our data shed light to the molecular mechanisms responsible for the anticancer activity of lumican in invasive breast cancer.