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Open Access 01-12-2019 | Epigenetics | Research article

The histone methyltransferase WHSC1 is regulated by EZH2 and is important for ovarian clear cell carcinoma cell proliferation

Authors: Machiko Kojima, Kenbun Sone, Katsutoshi Oda, Ryuji Hamamoto, Syuzo Kaneko, Shinya Oki, Asako Kukita, Hidenori Machino, Harunori Honjoh, Yoshiko Kawata, Tomoko Kashiyama, Kayo Asada, Michihiro Tanikawa, Mayuyo Mori-Uchino, Tetsushi Tsuruga, Kazunori Nagasaka, Yoko Matsumoto, Osamu Wada-Hiraike, Yutaka Osuga, Tomoyuki Fujii

Published in: BMC Cancer | Issue 1/2019

Abstract

Background

Wolf-Hirschhorn syndrome candidate gene-1 (WHSC1), a histone methyltransferase, has been found to be upregulated and its expression to be correlated with expression of enhancer of zeste homolog 2 (EZH2) in several cancers. In this study, we evaluated the role of WHSC1 and its therapeutic significance in ovarian clear cell carcinoma (OCCC).

Methods

First, we analyzed WHSC1 expression by quantitative PCR and immunohistochemistry using 23 clinical OCCC specimens. Second, the involvement of WHSC1 in OCCC cell proliferation was evaluated by MTT assays after siRNA-mediated WHSC1 knockdown. We also performed flow cytometry (FACS) to address the effect of WHSC1 on cell cycle. To examine the functional relationship between EZH2 and WHSC1, we knocked down EZH2 using siRNAs and checked the expression levels of WHSC1 and its histone mark H3K36m2 in OCCC cell lines. Finally, we checked WHSC1 expression after treatment with the selective inhibitor, GSK126.

Results

Both quantitative PCR and immunohistochemical analysis revealed that WHSC1 was significantly overexpressed in OCCC tissues compared with that in normal ovarian tissues. MTT assay revealed that knockdown of WHSC1 suppressed cell proliferation, and H3K36me2 levels were found to be decreased in immunoblotting. FACS revealed that WHSC1 knockdown affected the cell cycle. We also confirmed that WHSC1 expression was suppressed by EZH2 knockdown or inhibition, indicating that EZH2 is upstream of WHSC1 in OCCC cells.

Conclusions

WHSC1 overexpression induced cell growth and its expression is, at least in part, regulated by EZH2. Further functional analysis will reveal whether WHSC1 is a promising therapeutic target for OCCC.

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Title: The histone methyltransferase WHSC1 is regulated by EZH2 and is important for ovarian clear cell carcinoma cell proliferation
Authors: Machiko Kojima
Kenbun Sone
Katsutoshi Oda
Ryuji Hamamoto
Syuzo Kaneko
Shinya Oki
Asako Kukita
Hidenori Machino
Harunori Honjoh
Yoshiko Kawata
Tomoko Kashiyama
Kayo Asada
Michihiro Tanikawa
Mayuyo Mori-Uchino
Tetsushi Tsuruga
Kazunori Nagasaka
Yoko Matsumoto
Osamu Wada-Hiraike
Yutaka Osuga
Tomoyuki Fujii
Publication date: 01-12-2019
Publisher: BioMed Central
Keyword: Epigenetics
Published in: BMC Cancer / Issue 1/2019
Electronic ISSN: 1471-2407
DOI: https://doi.org/10.1186/s12885-019-5638-9

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