Department of Excellence: Department of Pharmacological and Biomolecular Sciences, Università degli Studi di Milano, via Balzaretti 9, 20133 Milan, Italy.
Department of Biomedical Sciences for Health, Università degli Studi di Milano, via Mangiagalli 31, 20133 Milan, Italy.
Cells. 2019 Feb 11;8(2):143. doi: 10.3390/cells8020143.
Three-dimensional (3D) cell cultures allow the mimic of functions of living tissues andprovide key information encoded in tissue architecture. Considered the pivotal role of epithelial-tomesenchymaltransition (EMT) in carcinoma progression, including prostate cancer (PCa), weaimed at investigating the effect of the 3D arrangement on the expression of some key markers ofEMT in cultured human prostate cancer (PCa) cells, to better understand PCa cell behavior. PC3 andDU145 PCa cells were cultured in RPMI cell culture medium either in 2D-monolayers or in 3Dspheroids.The main EMT markers E-cadherin, N-cadherin, α-smooth muscle actin (αSMA),vimentin, Snail, Slug, Twist and Zeb1 were evaluated by confocal microscopy, real-time PCR andWestern blot. Confocal microscopy revealed that E-cadherin was similarly expressed at the cellboundaries on the plasma membrane of PCa cells grown in 2D-monolayers, as well as in 3Dspheroids,but resulted up-regulated in 3D-spheroids, compared to 2D-monolayers, at the mRNAand protein level. Moreover, markers of the mesenchymal phenotype were expressed at very lowlevels in 3D-spheroids, suggesting important differences in the phenotype of PCa cells grown in 3Dspheroidsor in 2D-monolayers. Considered as a whole, our findings contribute to a clarification ofthe role of EMT in PCa and confirm that a 3D cell culture model could provide deeper insight intothe understanding of the biology of PCa.
三维（3D）细胞培养允许模拟活组织的功能，并提供组织架构中编码的关键信息。考虑到上皮-间充质转化（EMT）在包括前列腺癌（PCa）在内的癌进展中的关键作用，我们旨在研究 3D 排列对培养的人前列腺癌（PCa）细胞中一些 EMT 关键标志物表达的影响，以更好地理解 PCa 细胞的行为。PC3 和 DU145 PCa 细胞在 RPMI 细胞培养基中分别在 2D 单层或 3D 球体中培养。通过共聚焦显微镜、实时 PCR 和 Western blot 评估 EMT 的主要标志物 E-钙粘蛋白、N-钙粘蛋白、α-平滑肌肌动蛋白（αSMA）、波形蛋白、Snail、Slug、Twist 和 Zeb1。共聚焦显微镜显示，E-钙粘蛋白在 2D 单层培养的 PCa 细胞的质膜上的细胞边界处同样表达，并且在 3D 球体中也表达，但与 2D 单层相比，在 mRNA 和蛋白水平上均上调。此外，间充质表型标志物在 3D 球体中表达水平非常低，表明在 3D 球体中培养的 PCa 细胞与在 2D 单层中培养的 PCa 细胞的表型存在重要差异。总的来说，我们的发现有助于阐明 EMT 在 PCa 中的作用，并证实 3D 细胞培养模型可以更深入地了解 PCa 的生物学。
