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Differential detection of cytoplasmic Wilms tumor 1 expression by immunohistochemistry, western blotting and mRNA quantification

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Title: Differential detection of cytoplasmic Wilms tumor 1 expression by immunohistochemistry, western blotting and mRNA quantification
Authors: Maki, Takehiro Browse this author
Ikeda, Hiroaki Browse this author
Kuroda, Aki Browse this author
Kyogoku, Noriaki Browse this author
Yamamura, Yoshiyuki Browse this author
Tabata, Yukiko Browse this author
Abiko, Takehiro Browse this author
Tsuchikawa, Takahiro Browse this author →KAKEN DB
Hida, Yasuhiro Browse this author →KAKEN DB
Shichinohe, Toshiaki Browse this author →KAKEN DB
Tanaka, Eiichi Browse this author →KAKEN DB
Kaga, Kichizo Browse this author →KAKEN DB
Hatanaka, Kanako Browse this author
Matsuno, Yoshihiro Browse this author →KAKEN DB
Imai, Naoko Browse this author →KAKEN DB
Hirano, Satoshi Browse this author →KAKEN DB
Keywords: immunoblotting
reverse transcriptase polymerase chain reaction
Wilms Tumor 1 protein
Issue Date: Jan-2017
Publisher: Spandidos Publications
Journal Title: International journal of oncology
Volume: 50
Issue: 1
Start Page: 129
End Page: 140
Publisher DOI: 10.3892/ijo.2016.3786
PMID: 27922671
Abstract: Wilms tumor 1 (WT1) is considered to be a promising target of cancer treatment because it has been reported to be frequently expressed at high levels in various malignancies. Although WT1-targeted cancer treatment has been initiated, conclusive detection methods for WT1 are not established. The present study aimed to consolidate immunohistochemistry for WT1 with statistical basis. Transfected cells with forced WT1 expression yielded specific western blot bands and nuclear immunostaining; cytoplasmic immunostaining was not specifically recognized. Immunohistochemistry, western blotting, and quantitative reverse transcriptase-polymerase chain reaction were performed in 35 human cell lines using multiple WT1 antibodies and their results were quantified. Relationships among the quantified results were statistically analyzed; the nuclear immunostaining positively correlated with western blot bands and mRNA expression levels, whereas cytoplasmic immunostaining did not. These results indicate that nuclear immunostaining reflects WT1 expression but cytoplasmic immunostaining does not. The nuclear immunostaining was barely (3/541) observed in primary cancer of esophagus, bile duct, pancreas and lung. Although the present study has some limitations, the results indicate that the cytoplasmic immunostaining does not correlate with actual WT1 expression and prompts researchers to carefully evaluate target molecule expression in treatment of cancer.
Type: article
Appears in Collections:医学院・医学研究院 (Graduate School of Medicine / Faculty of Medicine) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 平野 聡

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