Comparative secretome analysis of cholangiocarcinoma cell line in three-dimensional culture

  • Authors:
    • Phanthakarn Tit-oon
    • Daranee Chokchaichamnankit
    • Amnart Khongmanee
    • Phannee Sawangareetrakul
    • Jisnuson Svasti
    • Chantragan Srisomsap
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  • Published online on: September 3, 2014     https://doi.org/10.3892/ijo.2014.2636
  • Pages: 2108-2116
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Abstract

Cholangiocarcinoma (CCA) is a lethal malignancy which occurs with relatively high incidence in Thailand. This cancer is often difficult to diagnose and associated with high mortality. The secretome, containing the secreted proteins from cells, are potentially useful as biomarkers of cancers. Since three-dimensional (3D) cell culture may mimic growth characteristics and microenvironment of solid tumors in vivo better than monolayer culture, we have developed culture of CCA in natural collagen-based scaffold, to enable analysis of the secretome by 2DE. Our results indicated that CCA growth in 3D environment alters cell shape significantly and enhances extracellular matrix deposition. Interestingly, more secreted proteins were detected from 3D culture compared to monolayer culture. Secretome analysis using 2DE coupled with LC-MS/MS demonstrated 10 secreted proteins uniquely found in 3D culture. Moreover, 25 proteins were enriched in 3D culture compared to monolayer culture, including 14-3-3 σ, triosephosphate isomerase, phosphoglycerate mutase 1, α-enolase, and L-plastin. Immunoblotting was used to confirm the presence of L-plastin in conditioned media of CCA and of hepatocellular carcinoma (HCC) cell lines. The results revealed that L-plastin, an actin bundling protein, was uniquely expressed only in the CCA cell line and could be a promising biomarker for differential diagnosis of CCA compared to HCC.
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November-2014
Volume 45 Issue 5

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

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Spandidos Publications style
Tit-oon P, Chokchaichamnankit D, Khongmanee A, Sawangareetrakul P, Svasti J and Srisomsap C: Comparative secretome analysis of cholangiocarcinoma cell line in three-dimensional culture. Int J Oncol 45: 2108-2116, 2014
APA
Tit-oon, P., Chokchaichamnankit, D., Khongmanee, A., Sawangareetrakul, P., Svasti, J., & Srisomsap, C. (2014). Comparative secretome analysis of cholangiocarcinoma cell line in three-dimensional culture. International Journal of Oncology, 45, 2108-2116. https://doi.org/10.3892/ijo.2014.2636
MLA
Tit-oon, P., Chokchaichamnankit, D., Khongmanee, A., Sawangareetrakul, P., Svasti, J., Srisomsap, C."Comparative secretome analysis of cholangiocarcinoma cell line in three-dimensional culture". International Journal of Oncology 45.5 (2014): 2108-2116.
Chicago
Tit-oon, P., Chokchaichamnankit, D., Khongmanee, A., Sawangareetrakul, P., Svasti, J., Srisomsap, C."Comparative secretome analysis of cholangiocarcinoma cell line in three-dimensional culture". International Journal of Oncology 45, no. 5 (2014): 2108-2116. https://doi.org/10.3892/ijo.2014.2636