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Prognostic significance of mesothelin expression in colorectal cancer disclosed by area-specific four-point tissue microarrays


Mesothelin (MSLN) is a cell surface glycoprotein present in many cancer types. Its expression is generally associated with an unfavorable prognosis. This study examined the prognostic significance of MSLN expression in different areas of individual colorectal cancers (CRCs) using tissue microarrays (TMAs) by enrolling 314 patients with stage II (T3–T4, N0, M0) CRCs. Using formalin-fixed paraffin-embedded tissue blocks from patients, TMA blocks were constructed. Tissue core specimens were obtained from submucosal invasive front (Fr-sm), subserosal invasive front (Fr-ss), central area (Ce), and rolled edge (Ro) of each tumor. Using these four-point TMA sets, MSLN expression was immunohistochemically surveyed. The area-specific prognostic significance of MSLN expression was evaluated. A deep learning convolutional neural network algorithm was used for imaging analysis and evaluating our judgment’s objectivity. MSLN staining ratio was positively correlated between the manual and machine-learning analyses (r = 0.71). The correlation coefficient between Ro and Ce, Ro and Fr-sm, and Ro and Fr-ss was r = 0.63, r = 0.54, and r = 0.61, respectively. Disease-specific survival curves for the MSLN-positive and MSLN-negative groups in Fr-sm, Fr-ss, and Ro were significantly different (five-year survival rates 88.1% and 95.5% (P = 0.024), 85.0 and 96.2% (P = 0.0087), 87.8 and 95.5% (P = 0.051), and 77.9 and 95.8% (P = 0.046) for Fr-sm, Fr-ss, Ce, and Ro, respectively). The analysis performed using area-specific four-point TMAs clearly demonstrated that MSLN expression in stage II CRC was relatively homogeneous within tumors. Additionally, high MSLN expression showed or tended to show unfavorable prognostic significance regardless of the tumor area.

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Confidence interval


Disease-specific survival


Hazard ratio


Receiver operating characteristic


Tissue microarray


Epithelial mesenchymal transition


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I.P.N. is the recipient of a Medical Research Scotland PhD Studentship awarded to P.D.C. Indica Labs, Inc. provided in kind resource.

Author information




TS and ES conceived and designed the experiments. TS and IPN performed the experiments. IPN performed the digital image analysis. TS, ES, and HT analyzed the histopathological data. TS, ES, and IPN drafted the manuscript. YK, TE, PDC, YK, and HU revised the manuscript. TS finalized the manuscript. All authors reviewed and approved the manuscript.

Corresponding author

Correspondence to Eiji Shinto.

Ethics declarations

The experiments reported here were performed in agreement with the Declaration of Helsinki principles and with the Ethics Committee of the National Defense Medical College Hospital, Tokorozawa, Japan. ES is the guarantor of this work and, as such, had full access to all of the data in the study and takes responsibility for the integrity of the data and the accuracy of the data analysis.

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The authors declare that they have no conflict of interest.

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Shiraishi, T., Shinto, E., Nearchou, I.P. et al. Prognostic significance of mesothelin expression in colorectal cancer disclosed by area-specific four-point tissue microarrays. Virchows Arch 477, 409–420 (2020).

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  • Mesothelin
  • Colorectal cancer
  • Tissue microarray
  • Immunohistochemistry
  • Artificial intelligence
  • Deep learning