title: Loss of histone demethylase KDM6B enhances aggressiveness of pancreatic cancer through downregulation of C/EBPα.
abstract: Genetic mutations in pancreatic ductal adenocarcinoma (PDAC) with critical roles have been well examined. The recent discovery of alterations in genes encoding histone modifiers suggests their possible roles in the complexity of cancer development. We previously reported loss of heterozygosity of the KDM6B gene, which encodes a histone demethylase for trimethylated histone H3 lysine 27, a repressive chromatin mark, in PDAC cells. In this study, we demonstrated that loss of KDM6B enhanced aggressiveness of PDAC cells. KDM6B has been regarded as a tumor suppressor that mediates oncogenic KRAS-induced senescence. Consistently, KDM6B was highly expressed in pancreatic precancerous lesions (pancreatic intraepithelial neoplasms); then, the expression decreased as the malignant grade progressed. We found that knockdown of KDM6B in PDAC cells promoted tumor sphere formation and increased peritoneal dissemination and liver metastasis in vivo. Microarray and chromatin immunoprecipitation analysis implicated CEBPA for aggressiveness induced by KDM6B knockdown. CEBPA knockdown recapitulated the phenotypic change of PDAC cells after KDM6B knockdown, which was reversed by forced expression of C/EBPα. Moreover, similar protein expression patterns of KDM6B and C/EBPα in human PDAC emphasized their functional correlation. Notably, pharmacological inhibition of the H3K27 methylase EZH2 in PDAC cells inhibited tumor sphere formation along with the upregulation of CEBPA expression, and this effect was impaired in KDM6B knockdown cells, highlighting the role for KDM6B in the activation of CEBPA. Together, our results propose a significant role for the KDM6B-C/EBPα axis in the PDAC phenotype.
authors.affiliation: Department of Gastroenterology, Department of Hematology and Oncology and Department of Pathology, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655, Japan, Division of Clinical Genome Research, The Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan, Hepato-Biliary-Pancreatic Surgery Division, Artificial Organ and Transplantation Division, Department of Surgery, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo , Bunkyo-ku, Tokyo, 113-8655, Japan and Yamanashi Prefectural Central Hospital, 1-1-1 Fujimi-cho, Kofu-shi, Yamanashi-Prefecture 400-8506, Japan.
authors.forename: Keisuke
authors.initials: K
authors.lastname: Yamamoto
authors.affiliation: Department of Gastroenterology, Department of Hematology and Oncology and Department of Pathology, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655, Japan, Division of Clinical Genome Research, The Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan, Hepato-Biliary-Pancreatic Surgery Division, Artificial Organ and Transplantation Division, Department of Surgery, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo , Bunkyo-ku, Tokyo, 113-8655, Japan and Yamanashi Prefectural Central Hospital, 1-1-1 Fujimi-cho, Kofu-shi, Yamanashi-Prefecture 400-8506, Japan ktate-tky@umin.ac.jp.
authors.forename: Keisuke
authors.initials: K
authors.lastname: Tateishi
authors.affiliation: Department of Gastroenterology, Department of Hematology and Oncology and Department of Pathology, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655, Japan, Division of Clinical Genome Research, The Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan, Hepato-Biliary-Pancreatic Surgery Division, Artificial Organ and Transplantation Division, Department of Surgery, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo , Bunkyo-ku, Tokyo, 113-8655, Japan and Yamanashi Prefectural Central Hospital, 1-1-1 Fujimi-cho, Kofu-shi, Yamanashi-Prefecture 400-8506, Japan.
authors.forename: Yotaro
authors.initials: Y
authors.lastname: Kudo
authors.affiliation: Department of Hematology and Oncology and.
authors.forename: Tomohiko
authors.initials: T
authors.lastname: Sato
authors.affiliation: Department of Gastroenterology, Department of Hematology and Oncology and Department of Pathology, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655, Japan, Division of Clinical Genome Research, The Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan, Hepato-Biliary-Pancreatic Surgery Division, Artificial Organ and Transplantation Division, Department of Surgery, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo , Bunkyo-ku, Tokyo, 113-8655, Japan and Yamanashi Prefectural Central Hospital, 1-1-1 Fujimi-cho, Kofu-shi, Yamanashi-Prefecture 400-8506, Japan.
authors.forename: Shinzo
authors.initials: S
authors.lastname: Yamamoto
authors.affiliation: Department of Gastroenterology, Department of Hematology and Oncology and Department of Pathology, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655, Japan, Division of Clinical Genome Research, The Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan, Hepato-Biliary-Pancreatic Surgery Division, Artificial Organ and Transplantation Division, Department of Surgery, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo , Bunkyo-ku, Tokyo, 113-8655, Japan and Yamanashi Prefectural Central Hospital, 1-1-1 Fujimi-cho, Kofu-shi, Yamanashi-Prefecture 400-8506, Japan.
authors.forename: Koji
authors.initials: K
authors.lastname: Miyabayashi
authors.affiliation: Department of Pathology, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655, Japan.
authors.forename: Keisuke
authors.initials: K
authors.lastname: Matsusaka
authors.affiliation: Department of Gastroenterology, Department of Hematology and Oncology and Department of Pathology, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655, Japan, Division of Clinical Genome Research, The Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan, Hepato-Biliary-Pancreatic Surgery Division, Artificial Organ and Transplantation Division, Department of Surgery, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo , Bunkyo-ku, Tokyo, 113-8655, Japan and Yamanashi Prefectural Central Hospital, 1-1-1 Fujimi-cho, Kofu-shi, Yamanashi-Prefecture 400-8506, Japan.
authors.forename: Yoshinari
authors.initials: Y
authors.lastname: Asaoka
authors.affiliation: Department of Gastroenterology, Department of Hematology and Oncology and Department of Pathology, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655, Japan, Division of Clinical Genome Research, The Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan, Hepato-Biliary-Pancreatic Surgery Division, Artificial Organ and Transplantation Division, Department of Surgery, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo , Bunkyo-ku, Tokyo, 113-8655, Japan and Yamanashi Prefectural Central Hospital, 1-1-1 Fujimi-cho, Kofu-shi, Yamanashi-Prefecture 400-8506, Japan.
authors.forename: Hideaki
authors.initials: H
authors.lastname: Ijichi
authors.affiliation: Department of Gastroenterology, Department of Hematology and Oncology and Department of Pathology, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655, Japan, Division of Clinical Genome Research, The Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan, Hepato-Biliary-Pancreatic Surgery Division, Artificial Organ and Transplantation Division, Department of Surgery, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo , Bunkyo-ku, Tokyo, 113-8655, Japan and Yamanashi Prefectural Central Hospital, 1-1-1 Fujimi-cho, Kofu-shi, Yamanashi-Prefecture 400-8506, Japan.
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Hirata
authors.affiliation: Department of Gastroenterology, Department of Hematology and Oncology and Department of Pathology, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655, Japan, Division of Clinical Genome Research, The Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan, Hepato-Biliary-Pancreatic Surgery Division, Artificial Organ and Transplantation Division, Department of Surgery, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo , Bunkyo-ku, Tokyo, 113-8655, Japan and Yamanashi Prefectural Central Hospital, 1-1-1 Fujimi-cho, Kofu-shi, Yamanashi-Prefecture 400-8506, Japan.
authors.forename: Motoyuki
authors.initials: M
authors.lastname: Otsuka
authors.affiliation: Department of Gastroenterology, Department of Hematology and Oncology and Department of Pathology, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655, Japan, Division of Clinical Genome Research, The Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan, Hepato-Biliary-Pancreatic Surgery Division, Artificial Organ and Transplantation Division, Department of Surgery, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo , Bunkyo-ku, Tokyo, 113-8655, Japan and Yamanashi Prefectural Central Hospital, 1-1-1 Fujimi-cho, Kofu-shi, Yamanashi-Prefecture 400-8506, Japan.
authors.forename: Yousuke
authors.initials: Y
authors.lastname: Nakai
authors.affiliation: Department of Gastroenterology, Department of Hematology and Oncology and Department of Pathology, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655, Japan, Division of Clinical Genome Research, The Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan, Hepato-Biliary-Pancreatic Surgery Division, Artificial Organ and Transplantation Division, Department of Surgery, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo , Bunkyo-ku, Tokyo, 113-8655, Japan and Yamanashi Prefectural Central Hospital, 1-1-1 Fujimi-cho, Kofu-shi, Yamanashi-Prefecture 400-8506, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Isayama
authors.affiliation: Division of Clinical Genome Research, The Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan.
authors.forename: Tsuneo
authors.initials: T
authors.lastname: Ikenoue
authors.affiliation: Department of Hematology and Oncology and.
authors.forename: Mineo
authors.initials: M
authors.lastname: Kurokawa
authors.affiliation: Department of Pathology, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655, Japan.
authors.forename: Masashi
authors.initials: M
authors.lastname: Fukayama
authors.affiliation: Hepato-Biliary-Pancreatic Surgery Division, Artificial Organ and Transplantation Division, Department of Surgery, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo , Bunkyo-ku, Tokyo, 113-8655, Japan and.
authors.forename: Norihiro
authors.initials: N
authors.lastname: Kokudo
authors.affiliation: Yamanashi Prefectural Central Hospital, 1-1-1 Fujimi-cho, Kofu-shi, Yamanashi-Prefecture 400-8506, Japan.
authors.forename: Masao
authors.initials: M
authors.lastname: Omata
authors.affiliation: Department of Gastroenterology, Department of Hematology and Oncology and Department of Pathology, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655, Japan, Division of Clinical Genome Research, The Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan, Hepato-Biliary-Pancreatic Surgery Division, Artificial Organ and Transplantation Division, Department of Surgery, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo , Bunkyo-ku, Tokyo, 113-8655, Japan and Yamanashi Prefectural Central Hospital, 1-1-1 Fujimi-cho, Kofu-shi, Yamanashi-Prefecture 400-8506, Japan.
authors.forename: Kazuhiko
authors.initials: K
authors.lastname: Koike
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics* / Q000473:pathology; D022762:CCAAT-Enhancer-Binding Proteins / Q000096:biosynthesis* / Q000235:genetics; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000235:genetics*; D000071221:Enhancer of Zeste Homolog 2 Protein; D015972:Gene Expression Regulation, Neoplastic; D055785:Gene Knockdown Techniques; D056466:Histone Demethylases / Q000235:genetics; D006657:Histones / Q000235:genetics; D006801:Humans; D056484:Jumonji Domain-Containing Histone Demethylases / Q000037:antagonists & inhibitors / Q000235:genetics*; D010190:Pancreatic Neoplasms / Q000235:genetics* / Q000473:pathology; D063151:Polycomb Repressive Complex 2 / Q000235:genetics; D011518:Proto-Oncogene Proteins / Q000235:genetics; D016283:Proto-Oncogene Proteins p21(ras); D018631:ras Proteins / Q000235:genetics
chemical_list: D022762:CCAAT-Enhancer-Binding Proteins; C515848:CEBPA protein, human; D006657:Histones; C117307:KRAS protein, human; D011518:Proto-Oncogene Proteins; D056466:Histone Demethylases; D056484:Jumonji Domain-Containing Histone Demethylases; C524220:KDM6B protein, human; C498409:EZH2 protein, human; D000071221:Enhancer of Zeste Homolog 2 Protein; D063151:Polycomb Repressive Complex 2; D016283:Proto-Oncogene Proteins p21(ras); D018631:ras Proteins