Horwitz Vered, Davidson Ben, Stern Dganit, Tropé Claes G, Tavor Re'em Tali, Reich Reuven
Institute of Drug Research, School of Pharmacy, Faculty of Medicine, The Hebrew University of Jerusalem, Jerusalem, Israel.
Department of Pharmacology, Israel Institute for Biological Research, Ness Ziona, Israel.
PLoS One. 2016 Sep 13;11(9):e0162502. doi: 10.1371/journal.pone.0162502. eCollection 2016.
Ezrin and p130Cas are structural proteins with an important role in signaling pathways and have been shown to promote cancer dissemination. We previously reported on overexpression of both ezrin and p130Cas in breast carcinoma effusions compared to primary carcinomas. Since ovarian and breast carcinomas share the ability to disseminate by forming malignant effusions, we sought to study the role of these molecules in ovarian carcinoma (OC).
OC cell lines were cultured in two different 3-dimensional conditions, on alginate scaffolds and as spheroids, which served as models for solid tumor and malignant effusions, respectively. shRNA was used to reduce protein expression in the cells. The malignant potential was evaluated by chemo-invasion assay, branching capacity on Matrigel and rate of proliferation. Subsequently, clinical specimens of high-grade serous carcinoma effusions, ovarian tumors and solid metastases were analyzed for ezrin and p130Cas expression.
Higher ezrin expression was found in cells composing the spheroids compared to their counterparts cultured on alginate scaffold and in clinical samples of malignant effusions compared to solid tumors. In addition, reduced Ezrin expression impaired the invasion ability and the branching capacity of OC cells to a greater extent than reduced p130Cas expression. However, ezrin and p130Cas expression in effusions was unrelated to clinical outcome.
The 3-dimensional cell cultures were found to mimic the different tumor sites and be applicable as a model. The in vitro results concur with the clinical specimen analysis, suggesting that in OC, the role of ezrin in disease progression is more pronounced than that of p130Cas.
埃兹蛋白（ezrin）和p130Cas是在信号通路中起重要作用的结构蛋白，已被证明可促进癌症扩散。我们之前报道过，与原发性乳腺癌相比，ezrin和p130Cas在乳腺癌胸水中均有过表达。由于卵巢癌和乳腺癌都具有通过形成恶性胸水进行扩散的能力，我们试图研究这些分子在卵巢癌（OC）中的作用。
将OC细胞系分别培养在两种不同的三维条件下，一种是在藻酸盐支架上，另一种是形成球体，分别作为实体瘤和恶性胸水的模型。使用短发夹RNA（shRNA）降低细胞中的蛋白表达。通过化学侵袭试验、在基质胶上的分支能力和增殖率评估恶性潜能。随后，分析高级别浆液性癌胸水、卵巢肿瘤和实体转移灶的临床标本中ezrin和p130Cas的表达情况。
与在藻酸盐支架上培养的细胞相比，组成球体的细胞中ezrin表达更高；与实体瘤相比，恶性胸水的临床样本中ezrin表达更高。此外，与降低p130Cas表达相比，降低Ezrin表达对OC细胞侵袭能力和分支能力的损害更大。然而，胸水中ezrin和p130Cas的表达与临床结局无关。
发现三维细胞培养可模拟不同的肿瘤部位，可作为一种模型。体外实验结果与临床标本分析结果一致，表明在OC中，ezrin在疾病进展中的作用比p130Cas更显著。
