AIM: To analyze the effect of three-dimensional (3D)-arrangement on the expression of epithelial-to-mesenchymal transition markers in pancreatic adenocarcinoma (PDAC) cells. METHODS: HPAF-II, HPAC, and PL45 PDAC cells were cultured in either 2D-monolayers or 3D-spheroids. Ultrastructure was analyzed by transmission electron microscopy. The expression of E-cadherin, β-catenin, N-cadherin, collagen type I (COL-I), vimentin, α-smooth muscle actin (αSMA), and podoplanin was assayed by confocal microscopy in cells cultured on 12-mm diameter round coverslips and in 3D-spheroids. Gene expression for E-cadherin, Snail, Slug, Twist, Zeb1, and Zeb2 was quantified by real-time PCR. E-cadherin protein level and its electrophoretic pattern were studied by Western blot in cell lysates obtained from cells grown in 2D-monolayers and 3D-spheroids. RESULTS: The E-cadherin/β-catenin complex was expressed in a similar way in plasma membrane cell boundaries in both 2D-monolayers and 3D-spheroids. E-cadherin increased in lysates obtained from 3D-spheroids, while cleavage fragments were more evident in 2D-monolayers. N-cadherin expression was observed in very few PDAC cells grown in 2D-monolayers, but was more evident in 3D-spheroids. Some cells expressing COL-I were observed in 3D-spheroids. Podoplanin, expressed in collectively migrating cells, and αSMA were similarly expressed in both experimental conditions. The concomitant maintenance of the E-cadherin/β-catenin complex at cell boundaries supports the hypothesis of a collective migration for these cells, which is consistent with podoplanin expression. CONCLUSION: We show that a 3D-cell culture model could provide deeper insight into understanding the biology of PDAC and allow for the detection of marked differences in the phenotype of PDAC cells grown in 3D-spheroids.
Epithelial-to-mesenchymal transition in pancreatic ductal adenocarcinoma: Characterization in a 3D-cell culture model / N. Gagliano, G. Celesti, L. Tacchini, S. Pluchino, C. Sforza, M. Rasile, V. Valerio, L. Laghi, V. Conte, P. Procacci. - In: WORLD JOURNAL OF GASTROENTEROLOGY. - ISSN 1007-9327. - 22:18(2016 May 14), pp. 4466-4483.
|Titolo:||Epithelial-to-mesenchymal transition in pancreatic ductal adenocarcinoma: Characterization in a 3D-cell culture model|
GAGLIANO, NICOLETTA (Primo)
CELESTI, GIUSEPPE (Secondo)
CONTE, VINCENZO (Penultimo)
PROCACCI, PATRIZIA (Ultimo)
|Parole Chiave:||Epithelial-to-mesenchymal transition; E-cadherin; 3D-spheroids; Podoplanin; Pancreatic ductal adenocarcinoma|
|Settore Scientifico Disciplinare:||Settore BIO/17 - Istologia|
Settore BIO/16 - Anatomia Umana
|Data di pubblicazione:||14-mag-2016|
|Digital Object Identifier (DOI):||http://dx.doi.org/10.3748/wjg.v22.i18.4466|
|Appare nelle tipologie:||01 - Articolo su periodico|