title: Immunoreactivity of Wnt5a, Fzd2, Fzd6, and Ryk in glioblastoma: evaluative methodology for DAB chromogenic immunostaining.
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.
authors.affiliation: Department of Neurosurgery, Graduate School of Medical and Dental Sciences, Kagoshima University, 8-35-1 Sakuragaoka, Kagoshima, Kagoshima, 890-8520, Japan, hirahira@m2.kufm.kagoshima-u.ac.jp.
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Hirano
authors.forename: Hajime
authors.initials: H
authors.lastname: Yonezawa
authors.forename: Shunji
authors.initials: S
authors.lastname: Yunoue
authors.forename: Mika
authors.initials: M
authors.lastname: Habu
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Uchida
authors.forename: Takako
authors.initials: T
authors.lastname: Yoshioka
authors.forename: Shosei
authors.initials: S
authors.lastname: Kishida
authors.forename: Michiko
authors.initials: M
authors.lastname: Kishida
authors.forename: Tatsuki
authors.initials: T
authors.lastname: Oyoshi
authors.forename: Shingo
authors.initials: S
authors.lastname: Fujio
authors.forename: Sei
authors.initials: S
authors.lastname: Sugata
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Yamahata
authors.forename: Ryousuke
authors.initials: R
authors.lastname: Hanaya
authors.forename: Kazunori
authors.initials: K
authors.lastname: Arita
mesh_terms: D000368:Aged; D001932:Brain Neoplasms / Q000235:genetics* / Q000401:mortality / Q000473:pathology*; D005260:Female; D051157:Frizzled Receptors / Q000032:analysis*; D005909:Glioblastoma / Q000235:genetics* / Q000401:mortality / Q000473:pathology*; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D008297:Male; D008875:Middle Aged; D011379:Prognosis; D016016:Proportional Hazards Models; D011518:Proto-Oncogene Proteins / Q000032:analysis*; D020794:Receptor Protein-Tyrosine Kinases / Q000032:analysis*; D015996:Survival Rate; D051153:Wnt Proteins / Q000032:analysis*; D000071818:Wnt-5a Protein
chemical_list: C489742:FZD2 protein, human; C484687:FZD6 protein, human; D051157:Frizzled Receptors; D011518:Proto-Oncogene Proteins; C085389:WNT5A protein, human; D051153:Wnt Proteins; D000071818:Wnt-5a Protein; C080836:RYK protein, human; D020794:Receptor Protein-Tyrosine Kinases