EZH1/2 inhibition augments the anti-tumor effects of sorafenib in hepatocellular carcinoma

Yuko Kusakabe; Tetsuhiro Chiba; Motohiko Oshima; Shuhei Koide; Ola Rizq; Kazumasa Aoyama; Junjie Ao; Tatsuya Kaneko; Hiroaki Kanzaki; Kengo Kanayama; Takahiro Maeda; Tomoko Saito; Ryo Nakagawa; Kazufumi Kobayashi; Soichiro Kiyono; Masato Nakamura; Sadahisa Ogasawara; Eiichiro Suzuki; Shingo Nakamoto; Shin Yasui; Rintaro Mikata; Ryosuke Muroyama; Tatsuo Kanda; Hitoshi Maruyama; Jun Kato; Naoya Mimura; Anqi Ma; Jian Jin; Yoh Zen; Masayuki Otsuka; Atsushi Kaneda; Atsushi Iwama; Naoya Kato

doi:10.1038/s41598-021-00889-0

EZH1/2 inhibition augments the anti-tumor effects of sorafenib in hepatocellular carcinoma

Sci Rep. 2021 Nov 1;11(1):21396. doi: 10.1038/s41598-021-00889-0.

Authors

Yuko Kusakabe¹, Tetsuhiro Chiba², Motohiko Oshima^{3

4}, Shuhei Koide^{3

4}, Ola Rizq^{3

4}, Kazumasa Aoyama⁴, Junjie Ao¹, Tatsuya Kaneko¹, Hiroaki Kanzaki¹, Kengo Kanayama¹, Takahiro Maeda¹, Tomoko Saito¹, Ryo Nakagawa¹, Kazufumi Kobayashi¹, Soichiro Kiyono¹, Masato Nakamura¹, Sadahisa Ogasawara¹, Eiichiro Suzuki¹, Shingo Nakamoto¹, Shin Yasui¹, Rintaro Mikata¹, Ryosuke Muroyama⁵, Tatsuo Kanda⁶, Hitoshi Maruyama⁷, Jun Kato¹, Naoya Mimura⁸, Anqi Ma^{9

10

11}, Jian Jin^{9

10

11}, Yoh Zen¹², Masayuki Otsuka¹³, Atsushi Kaneda¹⁴, Atsushi Iwama^{3

4}, Naoya Kato¹

Affiliations

¹ Department of Gastroenterology, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba, 260-8670, Japan.
² Department of Gastroenterology, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba, 260-8670, Japan. chibat@chiba-u.jp.
³ Division of Stem Cell and Molecular Medicine, Center for Stem Cell Biology and Regenerative Medicine, The Institute of Medical Science, The University of Tokyo, 4-6-1 Shirokenadai, Minato-ku, Tokyo, 108-8639, Japan.
⁴ Department of Cellular and Molecular Medicine, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba, 260-8670, Japan.
⁵ Department of Molecular Virology, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba, 260-8670, Japan.
⁶ Department of Gastroenterology and Hepatology, Nihon University School of Medicine, 30-1 Oyaguchi-Kamicho, Itabashi-ku, Tokyo, 173-8610, Japan.
⁷ Department of Gastroenterology, Juntendo University School of Medicine, 2-1-1 Hongo, Bunkyo-ku, Tokyo, 113-08421, Japan.
⁸ Department of Transfusion Medicine and Cell Therapy, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba, 260-8670, Japan.
⁹ Mount Sinai Center for Therapeutics Discovery, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA.
¹⁰ Department of Pharmacological Sciences, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA.
¹¹ Department of Oncological Sciences, Tisch Cancer Institute, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA.
¹² Institute of Liver Studies, King's College Hospital, London, UK.
¹³ Department of General Surgery, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba, 260-8670, Japan.
¹⁴ Department of Molecular Oncology, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba, 260-8670, Japan.

Abstract

Both EZH2 and its homolog EZH1 function as histone H3 Lysine 27 (H3K27) methyltransferases and repress the transcription of target genes. Dysregulation of H3K27 trimethylation (H3K27me3) plays an important role in the development and progression of cancers such as hepatocellular carcinoma (HCC). This study investigated the relationship between the expression of EZH1/2 and the level of H3K27me3 in HCC. Additionally, the role of EZH1/2 in cell growth, tumorigenicity, and resistance to sorafenib were also analyzed. Both the lentiviral knockdown and the pharmacological inhibition of EZH1/2 (UNC1999) diminished the level of H3K27me3 and suppressed cell growth in liver cancer cells, compared with EZH1 or EZH2 single knockdown. Although a significant association was observed between EZH2 expression and H3K27me3 levels in HCC samples, overexpression of EZH1 appeared to contribute to enhanced H3K27me3 levels in some EZH2^lowH3K27me3^high cases. Akt suppression following sorafenib treatment resulted in an increase of the H3K27me3 levels through a decrease in EZH2 phosphorylation at serine 21. The combined use of sorafenib and UNC1999 exhibited synergistic antitumor effects in vitro and in vivo. Combination treatment canceled the sorafenib-induced enhancement in H3K27me3 levels, indicating that activation of EZH2 function is one of the mechanisms of sorafenib-resistance in HCC. In conclusion, sorafenib plus EZH1/2 inhibitors may comprise a novel therapeutic approach in HCC.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Aged
Animals
Antineoplastic Agents / therapeutic use*
Benzamides / therapeutic use*
Carcinoma, Hepatocellular / genetics
Carcinoma, Hepatocellular / therapy*
Cell Line, Tumor
Down-Regulation / drug effects
Enhancer of Zeste Homolog 2 Protein / antagonists & inhibitors*
Enhancer of Zeste Homolog 2 Protein / genetics
Female
Genetic Therapy
Humans
Indazoles / therapeutic use*
Liver Neoplasms / genetics
Liver Neoplasms / therapy*
Male
Mice
Mice, SCID
Middle Aged
Piperazines / therapeutic use*
Polycomb Repressive Complex 2 / antagonists & inhibitors*
Polycomb Repressive Complex 2 / genetics
Pyridones / therapeutic use*
Sorafenib / therapeutic use*

Substances

Antineoplastic Agents
Benzamides
Indazoles
Piperazines
Pyridones
UNC1999
Sorafenib
EZH1 protein, human
EZH2 protein, human
Enhancer of Zeste Homolog 2 Protein
Polycomb Repressive Complex 2

Abstract

Publication types

MeSH terms

Substances

Grants and funding