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Hepatic stellate cells promote upregulation of epithelial cell adhesion molecule and epithelial-mesenchymal transition in hepatic cancer cells

  • Authors:
    • Teruya Nagahara
    • Hidenori Shiraha
    • Hiroaki Sawahara
    • Daisuke Uchida
    • Yasuto Takeuchi
    • Masaya Iwamuro
    • Junro Kataoka
    • Shigeru Horiguchi
    • Takeshi Kuwaki
    • Hideki Onishi
    • Shinichiro Nakamura
    • Akinobu Takaki
    • Kazuhiro Nouso
    • Kazuhide Yamamoto
  • View Affiliations / Copyright

    Affiliations: Department of Gastroenterology and Hepatology, Okayama University Graduate School of Medicine, Shikata-cho, Kita-ku, Okayama 700-8558, Japan
    Copyright: © Nagahara et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].
  • Pages: 1169-1177
    |
    Published online on: July 13, 2015
       https://doi.org/10.3892/or.2015.4126
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Abstract

Microenvironment plays an important role in epithelial-mesenchymal transition (EMT) and stemness of cells in hepatocellular carcinoma (HCC). Epithelial cell adhesion molecule (EpCAM) is known as a tumor stemness marker of HCC. To investigate the relationship between microenvironment and stemness, we performed an in vitro co-culture assay. Four HCC cell lines (HepG2, Hep3B, HuH-7 and PLC/PRF/5) were co-cultured with the TWNT-1 immortalized hepatic stellate cells (HSCs), which create a microenvironment with HCC. Cell proliferation ability was analyzed by flow cytometry (FCM) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, while migration ability was assessed by a wound healing assay. Expression of EpCAM was analyzed by immunoblotting and FCM. HCC cell lines were co-cultured with TWNT-1 treated with small interfering RNA (siRNA) for TGF-β and HB-EGF; we then analyzed proliferation, migration ability and protein expression using the methods described above. Proliferation ability was unchanged in HCC cell lines co-cultured with TWNT-1. Migration ability was increased in HCC cell lines (HepG2, Hep3B, HuH-7 and PLC/PRF/5) directly (216.2±67.0, 61.0±22.0, 124.0±66.2 and 51.5±40.3%) and indirectly (102.5±22.0, 84.6±30.9, 86.1±25.7 and 73.9±29.7%) co-cultured with TWNT-1 compared with the HCC uni-culture. Immunoblot analysis revealed increased EpCAM expression in the HCC cell lines co-cultured with TWNT-1. Flow cytometry revealed that the population of E-cadherin-/N-cadherin+ and EpCAM-positive cells increased and accordingly, EMT and stemness in the HCC cell line were activated. These results were similar in the directly and indirectly co-cultured samples, indicating that humoral factors were at play. Conversely, HCC cell lines co-cultured with siRNA‑treated TWNT-1 showed decreased migration ability, a decreased population of EpCAM-positive and E-cadherin-/N-cadherin+ cells. Taken together, humoral factors secreted from TWNT-1 promote upregulation of EpCAM and EMT in hepatic cancer cells.
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Copy and paste a formatted citation
Spandidos Publications style
Nagahara T, Shiraha H, Sawahara H, Uchida D, Takeuchi Y, Iwamuro M, Kataoka J, Horiguchi S, Kuwaki T, Onishi H, Onishi H, et al: Hepatic stellate cells promote upregulation of epithelial cell adhesion molecule and epithelial-mesenchymal transition in hepatic cancer cells. Oncol Rep 34: 1169-1177, 2015.
APA
Nagahara, T., Shiraha, H., Sawahara, H., Uchida, D., Takeuchi, Y., Iwamuro, M. ... Yamamoto, K. (2015). Hepatic stellate cells promote upregulation of epithelial cell adhesion molecule and epithelial-mesenchymal transition in hepatic cancer cells. Oncology Reports, 34, 1169-1177. https://doi.org/10.3892/or.2015.4126
MLA
Nagahara, T., Shiraha, H., Sawahara, H., Uchida, D., Takeuchi, Y., Iwamuro, M., Kataoka, J., Horiguchi, S., Kuwaki, T., Onishi, H., Nakamura, S., Takaki, A., Nouso, K., Yamamoto, K."Hepatic stellate cells promote upregulation of epithelial cell adhesion molecule and epithelial-mesenchymal transition in hepatic cancer cells". Oncology Reports 34.3 (2015): 1169-1177.
Chicago
Nagahara, T., Shiraha, H., Sawahara, H., Uchida, D., Takeuchi, Y., Iwamuro, M., Kataoka, J., Horiguchi, S., Kuwaki, T., Onishi, H., Nakamura, S., Takaki, A., Nouso, K., Yamamoto, K."Hepatic stellate cells promote upregulation of epithelial cell adhesion molecule and epithelial-mesenchymal transition in hepatic cancer cells". Oncology Reports 34, no. 3 (2015): 1169-1177. https://doi.org/10.3892/or.2015.4126
Copy and paste a formatted citation
x
Spandidos Publications style
Nagahara T, Shiraha H, Sawahara H, Uchida D, Takeuchi Y, Iwamuro M, Kataoka J, Horiguchi S, Kuwaki T, Onishi H, Onishi H, et al: Hepatic stellate cells promote upregulation of epithelial cell adhesion molecule and epithelial-mesenchymal transition in hepatic cancer cells. Oncol Rep 34: 1169-1177, 2015.
APA
Nagahara, T., Shiraha, H., Sawahara, H., Uchida, D., Takeuchi, Y., Iwamuro, M. ... Yamamoto, K. (2015). Hepatic stellate cells promote upregulation of epithelial cell adhesion molecule and epithelial-mesenchymal transition in hepatic cancer cells. Oncology Reports, 34, 1169-1177. https://doi.org/10.3892/or.2015.4126
MLA
Nagahara, T., Shiraha, H., Sawahara, H., Uchida, D., Takeuchi, Y., Iwamuro, M., Kataoka, J., Horiguchi, S., Kuwaki, T., Onishi, H., Nakamura, S., Takaki, A., Nouso, K., Yamamoto, K."Hepatic stellate cells promote upregulation of epithelial cell adhesion molecule and epithelial-mesenchymal transition in hepatic cancer cells". Oncology Reports 34.3 (2015): 1169-1177.
Chicago
Nagahara, T., Shiraha, H., Sawahara, H., Uchida, D., Takeuchi, Y., Iwamuro, M., Kataoka, J., Horiguchi, S., Kuwaki, T., Onishi, H., Nakamura, S., Takaki, A., Nouso, K., Yamamoto, K."Hepatic stellate cells promote upregulation of epithelial cell adhesion molecule and epithelial-mesenchymal transition in hepatic cancer cells". Oncology Reports 34, no. 3 (2015): 1169-1177. https://doi.org/10.3892/or.2015.4126
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