Institute for Drug Research, School of Pharmacy, Faculty of Medicine, The Hebrew University of Jerusalem, 91120, Jerusalem, Israel.
Clin Exp Metastasis. 2012 Aug;29(6):527-40. doi: 10.1007/s10585-012-9468-2. Epub 2012 Apr 4.
CAS proteins and Ezrin, Radixin, Moesin (ERM) family members act as intracellular scaffolds and are involved in interactions with the cytoskeleton, respectively. Both protein families have previously been associated with metastasis and poor prognosis in cancer. Our group recently reported on the overexpression of EZR/VIL2 and BCAR1 and their protein products in breast carcinoma effusions compared to primary breast carcinoma. In the present study, the role of these two proteins was studied in semi-normal MCF10A cells and metastatic MDA-MB-231 breast carcinoma cells cultured in tri-dimensional (3-D) conditions that were hypothesized to reproduce the in vivo conditions of breast cancer metastasis. MCF10A cells formed spheroid-shaped colonies without any Matrigel invasion, while MDA-MB-231 cells displayed an invasive phenotype and showed satellite projections that bridged multiple cell colonies in 3-D culture. E-cadherin was expressed in MCF10A, but not in MDA-MB-231 cells. The temporal expression of ezrin and BCAR1/p130Cas at the mRNA and protein level differed in the two cell lines upon 3-D culturing on Matrigel. Upregulation of BCAR1/p130cas was observed in the transition of MDA-MB-231 from attached to detached culture. Silencing of Ezrin and p130Cas in MDA-MB-231 cells by short hairpin RNA resulted in decreased invasive potential, and p130Cas silencing further resulted in smaller spheroid/colony formation. Our data show that MCF10A and MDA-MB-231 cells differ in their ability to form spheroids, in expression of E-cadherin and in the expression of Ezrin and BCAR1/p130Cas in 3-D cultures on Matrigel, suggesting a role in tumor progression in breast carcinoma.
CAS 蛋白和 Ezrin、Radixin、Moesin（ERM）家族成员分别作为细胞内支架和参与与细胞骨架的相互作用。这两个蛋白家族以前都与癌症的转移和预后不良有关。我们的研究小组最近报道了在乳腺癌渗液中 EzR/VIL2 和 BCAR1 及其蛋白产物的过度表达，与原发性乳腺癌相比。在本研究中，研究了这两种蛋白质在半正常 MCF10A 细胞和在三维（3-D）条件下培养的转移性 MDA-MB-231 乳腺癌细胞中的作用，假设这些条件可以再现乳腺癌转移的体内条件。MCF10A 细胞形成了没有任何 Matrigel 侵袭的球形菌落，而 MDA-MB-231 细胞表现出侵袭表型，并显示出卫星突起，在 3-D 培养中桥接了多个细胞菌落。E-钙粘蛋白在 MCF10A 中表达，但在 MDA-MB-231 细胞中不表达。在 Matrigel 上进行 3-D 培养时，两种细胞系中 ezrin 和 BCAR1/p130Cas 的 mRNA 和蛋白水平的表达在时间上不同。在 MDA-MB-231 从附着到游离培养的转变中观察到 BCAR1/p130cas 的上调。在 MDA-MB-231 细胞中用短发夹 RNA 沉默 Ezrin 和 p130Cas 导致侵袭潜能降低，p130Cas 沉默进一步导致较小的球体/集落形成。我们的数据表明，MCF10A 和 MDA-MB-231 细胞在形成球体的能力、E-钙粘蛋白的表达以及在 Matrigel 上进行 3-D 培养时 Ezrin 和 BCAR1/p130Cas 的表达上存在差异，这表明它们在乳腺癌肿瘤进展中起作用。
