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

Maki Takehiro; Ikeda Hiroaki; Kuroda Aki; Kyogoku Noriaki; Yamamura Yoshiyuki; Tabata Yukiko; Abiko Takehiro; Tsuchikawa Takahiro; Hida Yasuhiro; Shichinohe Toshiaki; Tanaka Eiichi; Kaga Kichizo; Hatanaka Kanako; Matsuno Yoshihiro; Imai Naoko; Hirano Satoshi

Title	en Differential detection of cytoplasmic Wilms tumor 1 expression by immunohistochemistry, western blotting and mRNA quantification
Creator	en Maki, Takehiro en Ikeda, Hiroaki en Kuroda, Aki en Kyogoku, Noriaki en Yamamura, Yoshiyuki en Tabata, Yukiko en Abiko, Takehiro en Tsuchikawa, Takahiro NRID 1000050507572 en Hida, Yasuhiro NRID 1000030399919 en Shichinohe, Toshiaki NRID 1000070374353 en Tanaka, Eiichi NRID 1000060374279 en Kaga, Kichizo NRID 1000080224335 en Hatanaka, Kanako en Matsuno, Yoshihiro NRID 1000000199829 en Imai, Naoko NRID 1000030550975 en Hirano, Satoshi NRID 1000050322813
Accessrights	open access
Subject	Other en immunoblotting Other en immunohistochemistry Other en neoplasms Other en reverse transcriptase polymerase chain reaction Other en Wilms Tumor 1 protein NDC 490
Description	Abstract en 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.
Publisher	en Spandidos Publications
Date	Issued2017-01
Language	eng
Resource Type	journal article
Version Type	VoR
Identifier	HDL http://hdl.handle.net/2115/66405
Relation	isIdenticalTo DOI https://doi.org/10.3892/ijo.2016.3786 PMID 27922671
Journal	PISSN 1019-6439 en International journal of oncology Volume Number50 Issue Number1 Page Start129 Page End140
File	fulltext Maki2017IntJOncolVol50No1Pg129.pdf 1.18 MB (application/pdf) Issued2017-01
Oaidate	2023-07-26