title: Down-regulation of histone deacetylase 4, -5 and -6 as a mechanism of synergistic enhancement of apoptosis in human lung cancer cells treated with the combination of a synthetic retinoid, Am80 and green tea catechin.
abstract: (-)-Epigallocatechin gallate (EGCG), a green tea catechin, acts as a synergist with various anticancer drugs, including retinoids. Am80 is a synthetic retinoid with a different structure from all-trans-retinoic acid: Am80 is now clinically utilized as a new drug for relapsed and intractable acute promyelocytic leukemia patients. Our experiments showed that the combination of EGCG and Am80 synergistically induced both apoptosis in human lung cancer cell line PC-9 and up-regulated expressions of growth arrest and DNA damage-inducible gene 153 (GADD153), death receptor 5, and p21waf1genes in the cells. To understand the mechanisms of synergistic anticancer activity of the combination, we gave special attention to the lysine acetylation of proteins. Proteomic analysis using nanoLC-ESI-MS/MS revealed that PC-9 cells treated with the combination contained 331 acetylated proteins, while nontreated cells contained 553 acetylated proteins, and 59 acetylated proteins were found in both groups. Among them, the combination increased acetylated-p53 and acetylated-α-tubulin through reduction of histone deacetylase (HDAC) activity in cytosol fraction, although the levels of acetylation in histones H3 or H4 did not change, and the combination reduced protein levels of HDAC4, -5 and -6 by 20% to 80%. Moreover, we found that a specific inhibitor of HDAC4 and -5 strongly induced p21waf1gene expression, and that of HDAC6 induced both GADD153 and p21waf1gene expression, which resulted in apoptosis. All results demonstrate that EGCG in combination with Am80 changes levels of acetylation in nonhistone proteins via down-regulation of HDAC4, -5 and -6 and stimulates apoptotic induction.
authors.affiliation: Graduate School of Science and Engineering, Saitama University, Shimo-okubo 255, Sakura-ku, Saitama, Saitama 338-8570, Japan; Research Institute for Clinical Oncology, Saitama Cancer Center, Ina, Kitaadachi-gun, Saitama 362-0806, Japan. Electronic address: taki.oya56@gmail.com.
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Oya
authors.affiliation: Graduate School of Science and Engineering, Saitama University, Shimo-okubo 255, Sakura-ku, Saitama, Saitama 338-8570, Japan; Research Institute for Clinical Oncology, Saitama Cancer Center, Ina, Kitaadachi-gun, Saitama 362-0806, Japan. Electronic address: amondal@mail.saitama-u.ac.jp.
authors.forename: Anupom
authors.initials: A
authors.lastname: Mondal
authors.affiliation: Graduate School of Science and Engineering, Saitama University, Shimo-okubo 255, Sakura-ku, Saitama, Saitama 338-8570, Japan; Research Institute for Clinical Oncology, Saitama Cancer Center, Ina, Kitaadachi-gun, Saitama 362-0806, Japan. Electronic address: ewmedsci@gmail.com.
authors.forename: Anchalee
authors.initials: A
authors.lastname: Rawangkan
authors.affiliation: Graduate School of Science and Engineering, Saitama University, Shimo-okubo 255, Sakura-ku, Saitama, Saitama 338-8570, Japan; Research Institute for Clinical Oncology, Saitama Cancer Center, Ina, Kitaadachi-gun, Saitama 362-0806, Japan. Electronic address: birdvet_67@hotmail.com.
authors.forename: Sonthaya
authors.initials: S
authors.lastname: Umsumarng
authors.affiliation: Graduate School of Science and Engineering, Saitama University, Shimo-okubo 255, Sakura-ku, Saitama, Saitama 338-8570, Japan; Research Institute for Clinical Oncology, Saitama Cancer Center, Ina, Kitaadachi-gun, Saitama 362-0806, Japan. Electronic address: e-da@cancer-c.pref.saitama.jp.
authors.forename: Keisuke
authors.initials: K
authors.lastname: Iida
authors.affiliation: Faculty of Medicine, Saga University, Nabeshima, Saga 849-8501, Japan. Electronic address: TATSURO.WATANABE@ucdenver.edu.
authors.forename: Tatsuro
authors.initials: T
authors.lastname: Watanabe
authors.affiliation: Graduate School of Science and Engineering, Saitama University, Shimo-okubo 255, Sakura-ku, Saitama, Saitama 338-8570, Japan; Research Institute for Clinical Oncology, Saitama Cancer Center, Ina, Kitaadachi-gun, Saitama 362-0806, Japan. Electronic address: mikimo-k@t.vodafone.ne.jp.
authors.forename: Miki
authors.initials: M
authors.lastname: Kanno
authors.affiliation: Graduate School of Science and Engineering, Saitama University, Shimo-okubo 255, Sakura-ku, Saitama, Saitama 338-8570, Japan; Research Institute for Clinical Oncology, Saitama Cancer Center, Ina, Kitaadachi-gun, Saitama 362-0806, Japan. Electronic address: orikaorin@gmail.com.
authors.forename: Kaori
authors.initials: K
authors.lastname: Suzuki
authors.affiliation: Graduate School of Science and Engineering, Saitama University, Shimo-okubo 255, Sakura-ku, Saitama, Saitama 338-8570, Japan; Research Institute for Clinical Oncology, Saitama Cancer Center, Ina, Kitaadachi-gun, Saitama 362-0806, Japan. Electronic address: risk1103@gmail.com.
authors.forename: Zhenghao
authors.initials: Z
authors.lastname: Li
authors.affiliation: Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University (TMDU), Tokyo 101-0062, Japan. Electronic address: kage.chem@tmd.ac.jp.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Kagechika
authors.affiliation: Japan Pharmaceutical Information Center, Shibuya, Tokyo 150-0002, Japan. Electronic address: kshudo@abeam.ocn.ne.jp.
authors.forename: Koichi
authors.initials: K
authors.lastname: Shudo
authors.affiliation: Faculty of Medicine, Saga University, Nabeshima, Saga 849-8501, Japan. Electronic address: uv4h-fjk@asahi-net.or.jp.
authors.forename: Hirota
authors.initials: H
authors.lastname: Fujiki
authors.affiliation: Graduate School of Science and Engineering, Saitama University, Shimo-okubo 255, Sakura-ku, Saitama, Saitama 338-8570, Japan; Research Institute for Clinical Oncology, Saitama Cancer Center, Ina, Kitaadachi-gun, Saitama 362-0806, Japan. Electronic address: masami0306@mail.saitama-u.ac.jp.
authors.forename: Masami
authors.initials: M
authors.lastname: Suganuma
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D001565:Benzoates / Q000494:pharmacology*; D002289:Carcinoma, Non-Small-Cell Lung / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D002392:Catechin / Q000031:analogs & derivatives* / Q000494:pharmacology; D045744:Cell Line, Tumor; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000235:genetics; D015536:Down-Regulation / Q000187:drug effects; D004357:Drug Synergism; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D018922:HL-60 Cells; D000073864:Histone Deacetylase 6 / Q000037:antagonists & inhibitors / Q000378:metabolism; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology*; D006655:Histone Deacetylases / Q000378:metabolism; D006801:Humans; D008175:Lung Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D012097:Repressor Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D013662:Tea / Q000737:chemistry; D013764:Tetrahydronaphthalenes / Q000494:pharmacology*; D051742:Transcription Factor CHOP / Q000235:genetics
chemical_list: D001565:Benzoates; C493956:CDKN1A protein, human; D050759:Cyclin-Dependent Kinase Inhibitor p21; C497249:DDIT3 protein, human; D056572:Histone Deacetylase Inhibitors; D012097:Repressor Proteins; D013662:Tea; D013764:Tetrahydronaphthalenes; C061133:tamibarotene; D051742:Transcription Factor CHOP; D002392:Catechin; C045651:epigallocatechin gallate; C400300:HDAC4 protein, human; C404500:HDAC5 protein, human; C433811:HDAC6 protein, human; D000073864:Histone Deacetylase 6; D006655:Histone Deacetylases
keywords: GADD153; HDAC; Lysine acetylation; Retinoid; p21(waf1); p53