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C-ERC/mesothelin provokes lymphatic invasion of colorectal adenocarcinoma

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Abstract

Background

Lymph node metastasis is a key event of colorectal cancer (CRC) progression. Mesothelin is expressed in various types of malignant tumor and associated with an unfavorable prognosis. The full-length mesothelin (Full-ERC) is cleaved by protease into membrane-bound C-ERC/mesothelin and N-ERC/mesothelin which is secreted into the blood. The aim of this study was to examine the biological role of mesothelin in CRC by clinicopathological analysis and in vitro lymphatic invasion assay.

Methods

Ninety-one cases of CRC specimens were immunohistochemically examined and the localization of mesothelin in luminal membrane and/or cytoplasm was also evaluated. Lymphatic invasion assay was also performed using the human CRC cell line, WiDr, which was transfected with Full-, N- and C-ERC/mesothelin expression plasmids (Full-WiDr, N-WiDr and C-WiDr).

Results

Immunohistochemically, “luminal membrane positive” of mesothelin was identified in 37.4 %, and correlated with lymphatic permeation and lymph node metastasis, but not with patients’ prognosis. Interestingly, among the patients with lymph node metastasis (N = 38), “luminal membrane positive” of mesothelin significantly correlated with unfavorable patients’ outcome. In addition, lymphatic invasion assay revealed that Full-WiDr and C-WiDr more significantly invaded human lymphatic endothelial cells than the Mock-WiDr (P < 0.01).

Conclusion

The luminal membrane expression of mesothelin was associated with unfavorable prognosis of CRC patients with lymph node metastasis. Moreover, this is the first report to prove the biological function of C-ERC/mesothelin associated with lymphatic invasion of cancer in vitro.

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Acknowledgments

This research was supported by a Grant-in-Aid for Scientific Research (KAKENHI) and by the Japan Society for the Promotion of Science (JSPS) through the “Funding Program for World-Leading Innovative R&D on Science and Technology (FIRST Program),” initiated by the Council for Science and Technology Policy (CSTP).

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of General Surgery, Hokkaido University, Graduate School of Medicine, Sapporo, Japan

    Futoshi Kawamata, Shigenori Homma, Hirofumi Kamachi, Takahiro Einama, Norihiko Takahashi, Toshiya Kamiyama, Akinobu Taketomi & Satoru Todo

  2. Department of Cancer Pathology, Hokkaido University School of Medicine, Sapporo, Japan

    Yasutaka Kato, Masumi Tsuda & Shinya Tanaka

  3. Department of Research and Development, Immuno-Biological Laboratories Co. Ltd., Gunma, Japan

    Masahiro Maeda

  4. Department of Pathology and Oncology, Juntendo University School of Medicine, Tokyo, Japan

    Kazunori Kajino & Okio Hino

  5. Department of Translational Pathology, Hokkaido University, Graduate School of Medicine, Kita-ku, N 15, W 7, Sapporo, 060-8638, Japan

    Hiroshi Nishihara

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535_2013_773_MOESM1_ESM.tif

Supplemental Figure 1. Scheme of lymphatic invasion and adhesion assay. (a): Modified transwell invasion assays. 5 x 104 of GFP-expressing Full-WiDr, N-WiDr, C-WiDr or Mock-WiDr were loaded on the top of the upper side of transwell inserts which were covered with a confluent monolayer of hLEC, and invaded cells were counted on the bottom side of the inserts. (b): Modified adhesion assays. 1 x 105 of hLEC were plated on a 12-well dish and incubated for 3 days. After confirmation of hLEC in confluence and monolayer by microscope, 1 x 105 of GFP-expressing Full-WiDr, N-WiDr, C-WiDr or Mock-WiDr were resuspended and plated on the cell layer of hLEC. After the incubation at 37°C for 30 min, non-adherent or loosely attached cells were removed by gently washing the wells twice with PBS. The attached GFP-expressing cells (Full-WiDr, N-WiDr, C-WiDr and Mock-WiDr) on hLEC were counted using a fluorescence microscope. hLEC: human lymphatic endothelial cells (TIFF 102 kb)

535_2013_773_MOESM2_ESM.tif

Supplemental Figure 2. Expression of mesothelin and patients’ prognosis (a, c): “High-level expression” and “cytoplasmic expression” of mesothelin were not correlated with total patients’ prognosis (N = 91, P = 0.26, P = 0.31). (b, d): Among the patients with lymph node metastasis (N = 38), “high-level expression” and “cytoplasmic expression” of mesothelin were not correlated with patients’ prognosis (P = 0.73, P = 0.89) (TIFF 75 kb)

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Kawamata, F., Homma, S., Kamachi, H. et al. C-ERC/mesothelin provokes lymphatic invasion of colorectal adenocarcinoma. J Gastroenterol 49, 81–92 (2014). https://doi.org/10.1007/s00535-013-0773-6

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  • DOI: https://doi.org/10.1007/s00535-013-0773-6

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