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Immunoreactivity of Wnt5a, Fzd2, Fzd6, and Ryk in glioblastoma: evaluative methodology for DAB chromogenic immunostaining

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Abstract

The aim of this study was to determine the influence of Wnt5a and its receptors on the survival of glioblastoma patients and to determine reliable evaluation methods for immunohistochemistry. Diagnostic specimens from 41 histopathologically confirmed primary glioblastoma patients whose Gd-enhanced tumors had been totally removed were immunohistochemically stained for Wnt5a, Fzd2, Fzd6, and Ryk. The immunoreactivity was evaluated using the following methods: (A) grayscale optical density after color deconvolution, (B) percentage of stained cells, (C) density of stained cells, (D) staining amount (multiplication product of B and C), and (E) staining rank. The data sets of A to E were statistically evaluated by correlation matrix analysis and regression analysis. The influence of the expression of the markers on survival was analyzed using a proportional hazard model. The results of color deconvolution (A) were well correlated with the results of the staining rank (E). In the semiquantitative results (B, C, and D), the staining amount (D) tended to show a better correlation with results of color deconvolution (A). Among all data sets, color deconvolution (A) demonstrated the most preferable fit in a proportional hazard model, and the expression of Fzd2 and Fzd6 was associated with poor prognosis in glioblastoma patients.

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Acknowledgments

This study was aided by Grant-in-Aid no. 23592135 to Hirofumi Hirano by the Ministry of Education, Culture, Sports, Science and Technology-Japan. We thank Miss Mai Tokudome for the arrangement of specimens and clinical data.

Ethical standard

This retrospective study was endorsed by the ethics committee on clinical research at Kagoshima University Graduate School (no. 77 in 2010).

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Authors and Affiliations

  1. Department of Neurosurgery, Graduate School of Medical and Dental Sciences, Kagoshima University, 8-35-1 Sakuragaoka, Kagoshima, Kagoshima, 890-8520, Japan

    Hirofumi Hirano, Hajime Yonezawa, Shunji Yunoue, Mika Habu, Hiroyuki Uchida, Tatsuki Oyoshi, Shingo Fujio, Sei Sugata, Hitoshi Yamahata, Ryousuke Hanaya & Kazunori Arita

  2. Department of Molecular and Cellular Pathology, Field of Oncology, Graduate School of Medical and Dental Sciences, Kagoshima University, 8-35-1 Sakuragaoka, Kagoshima, Kagoshima, 890-8520, Japan

    Takako Yoshioka

  3. Department of Biochemistry and Genetics, Graduate School of Medical and Dental Sciences, Kagoshima University, 8-35-1 Sakuragaoka, Kagoshima, Kagoshima, 890-8520, Japan

    Shosei Kishida & Michiko Kishida

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  1. Hirofumi Hirano
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  2. Hajime Yonezawa
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  14. Kazunori Arita
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Correspondence to Hirofumi Hirano.

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Hirano, H., Yonezawa, H., Yunoue, S. et al. Immunoreactivity of Wnt5a, Fzd2, Fzd6, and Ryk in glioblastoma: evaluative methodology for DAB chromogenic immunostaining. Brain Tumor Pathol 31, 85–93 (2014). https://doi.org/10.1007/s10014-013-0153-1

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  • DOI: https://doi.org/10.1007/s10014-013-0153-1

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