Three-dimensional culture using a radial flow bioreactor induces matrix metalloprotease 7-mediated EMT-like process in tumor cells via TGFβ1/Smad pathway

  • Authors:
    • Shun-Ichi Shibata
    • Hideki Marushima
    • Tadashi Asakura
    • Tomokazu Matsuura
    • Homare Eda
    • Katsuhiko Aoki
    • Hiroshi Matsudaira
    • Kazu Ueda
    • Kiyoshi Ohkawa
  • View Affiliations

  • Published online on: May 1, 2009     https://doi.org/10.3892/ijo_00000272
  • Pages: 1433-1448
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Abstract

To confirm the usefulness of the radial flow type bioreactor (RFB) for a three-dimensional (3D) culture system, which provides a tissue architecture and molecular function mimicking the in vivo environment, molecular expression in the A431 human squamous carcinoma cell line during culture were analyzed under the physically different environments of 3D culture in the RFB, 2D culture in a monolayer as well as in nude mice. Time-dependent accumulation of autocrine transforming growth factor (TGF) β1 was found in spent culture media obtained only from 3D cultured A431 cancer cells, which grew well with a stratified-sheet morphology. Cells in the RFB overexpressed matrix metalloproteinase 7 (MMP7) and showed an increased release of soluble 80-kDa fragments of E-cadherin into the media time-dependently, resulting in the reduction of E-cadherin protein at the cell surface without down-regulation of the mRNA. β-Catenin and its nuclear partner, LEF1, were up-regulated and Wnt protein secretion was also accelerated. Additional up-regulation of the transcriptional factors, HMGA2 and down-stream Slug, was noted. TGFβ1-dependent, MMP7-mediated up-regulation of β-catenin/LEF1 signaling and TGFβ1-activated HMGA2 pathways consequently converged with Slug overexpression, due to disassembly and further repression of E-cadherin expression, which was reproducible in the epithelial mesenchymal transition process without any manipulation. Other transcriptional factors, Notch/HEY1 and NF-κB, were also up-regulated in 3D-cultured cells. These signals recruited molecules related to extracellular matrix-cell remodeling and angiogenesis. Expression of several representative molecules in the 3D cultured cells was parallel with that in xenotransplanted A431 tumor tissues in nude mice. 3D culture of tumor cells in the RFB is a useful tool for cancer experimental biology and evaluation of cancer therapeutic-like systems in nude mice.

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May 2009
Volume 34 Issue 5

Print ISSN: 1019-6439
Online ISSN:1791-2423

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Spandidos Publications style
Shibata S, Marushima H, Asakura T, Matsuura T, Eda H, Aoki K, Matsudaira H, Ueda K and Ohkawa K: Three-dimensional culture using a radial flow bioreactor induces matrix metalloprotease 7-mediated EMT-like process in tumor cells via TGFβ1/Smad pathway. Int J Oncol 34: 1433-1448, 2009
APA
Shibata, S., Marushima, H., Asakura, T., Matsuura, T., Eda, H., Aoki, K. ... Ohkawa, K. (2009). Three-dimensional culture using a radial flow bioreactor induces matrix metalloprotease 7-mediated EMT-like process in tumor cells via TGFβ1/Smad pathway. International Journal of Oncology, 34, 1433-1448. https://doi.org/10.3892/ijo_00000272
MLA
Shibata, S., Marushima, H., Asakura, T., Matsuura, T., Eda, H., Aoki, K., Matsudaira, H., Ueda, K., Ohkawa, K."Three-dimensional culture using a radial flow bioreactor induces matrix metalloprotease 7-mediated EMT-like process in tumor cells via TGFβ1/Smad pathway". International Journal of Oncology 34.5 (2009): 1433-1448.
Chicago
Shibata, S., Marushima, H., Asakura, T., Matsuura, T., Eda, H., Aoki, K., Matsudaira, H., Ueda, K., Ohkawa, K."Three-dimensional culture using a radial flow bioreactor induces matrix metalloprotease 7-mediated EMT-like process in tumor cells via TGFβ1/Smad pathway". International Journal of Oncology 34, no. 5 (2009): 1433-1448. https://doi.org/10.3892/ijo_00000272