title: Shikonin induces apoptosis of lung cancer cells via activation of FOXO3a/EGR1/SIRT1 signaling antagonized by p300.
abstract: Shikonin derivatives exert powerful cytotoxic effects including induction of apoptosis. Here, we demonstrate the cytotoxic efficacy of shikonin in vivo in xenograft models, which did not affect body weight as well as its reduction of cell viability in vitro using several non-small cell lung cancer (NSCLC) cell lines. We found that inhibition of AKT by shikonin activated the forkhead box (FOX)O3a/early growth response protein (EGR)1 signaling cascade and enhanced the expression of the target gene Bim, leading to apoptosis in lung cancer cells. Overexpression of wild-type or a constitutively active mutant of FOXO3a enhanced shikonin-induced Bim expression. The NAD+-dependent histone deacetylase sirtuin (SIRT)1 amplified the pro-apoptotic effect by deacetylating FOXO3a, which induced EGR1 binding to the Bim promoter and activated Bim expression. Meanwhile, PI3K/AKT activity was enhanced, whereas that of FOXO3a was reduced and p300 was upregulated by treatment with a sublethal dose of shikonin. FOXO3a acetylation was enhanced by p300 overexpression, while shikonin-induced Bim expression was suppressed by p300 overexpression, which promoted cell survival. FOXO3a acetylation was increased by p300 overexpression and treatment with SIRT1 inhibitor, improving cell survival. In addition, shikonin-induced FOXO3a nuclear localization was blocked by AKT activation and SIRT1 inhibition, which blocked Bim expression and conferred resistance to the cytotoxic effects of shikonin. The EGR1 increase induced by shikonin was restored by pretreatment with SIRT1 inhibitor. These results suggest that shikonin induces apoptosis in some lung cancer cells via activation of FOXO3a/EGR1/SIRT1 signaling, and that AKT and p300 negatively regulate this process via Bim upregulation.
authors.affiliation: Biomedical Translational Research Center, KRIBB, Daejeon 34141, Republic of Korea; Department of Biochemistry, Chungnam National University Medical School, Daejeon 301-747, Republic of Korea.
authors.forename: Yun-Ji
authors.initials: YJ
authors.lastname: Jeung
authors.affiliation: Biomedical Translational Research Center, KRIBB, Daejeon 34141, Republic of Korea; Functional Genomics, Korea University of Science and Technology (UST), Daejeon, 305-806, Republic of Korea.
authors.forename: Han-Gyeul
authors.initials: HG
authors.lastname: Kim
authors.affiliation: Biomedical Translational Research Center, KRIBB, Daejeon 34141, Republic of Korea.
authors.forename: Jiwon
authors.initials: J
authors.lastname: Ahn
authors.affiliation: Biomedical Translational Research Center, KRIBB, Daejeon 34141, Republic of Korea.
authors.forename: Ho-Joon
authors.initials: HJ
authors.lastname: Lee
authors.affiliation: Biomedical Translational Research Center, KRIBB, Daejeon 34141, Republic of Korea.
authors.forename: Sae-Bhom
authors.initials: SB
authors.lastname: Lee
authors.affiliation: Biomedical Translational Research Center, KRIBB, Daejeon 34141, Republic of Korea.
authors.forename: Misun
authors.initials: M
authors.lastname: Won
authors.affiliation: Biomedical Translational Research Center, KRIBB, Daejeon 34141, Republic of Korea.
authors.forename: Cho-Rock
authors.initials: CR
authors.lastname: Jung
authors.affiliation: Biomedical Translational Research Center, KRIBB, Daejeon 34141, Republic of Korea.
authors.forename: Joo-Young
authors.initials: JY
authors.lastname: Im
authors.affiliation: Biomedical Translational Research Center, KRIBB, Daejeon 34141, Republic of Korea.
authors.forename: Bo-Kyung
authors.initials: BK
authors.lastname: Kim
authors.affiliation: Department of Biochemistry, Chungnam National University Medical School, Daejeon 301-747, Republic of Korea.
authors.forename: Seung-Kiel
authors.initials: SK
authors.lastname: Park
authors.affiliation: Stem Cell Research Center, KRIBB, Daejeon 34141, Republic of Korea; Functional Genomics, Korea University of Science and Technology (UST), Daejeon, 305-806, Republic of Korea. Electronic address: mjson@kribb.re.kr.
authors.forename: Myung Jin
authors.initials: MJ
authors.lastname: Son
authors.affiliation: Biomedical Translational Research Center, KRIBB, Daejeon 34141, Republic of Korea; Functional Genomics, Korea University of Science and Technology (UST), Daejeon, 305-806, Republic of Korea. Electronic address: kschung@kribb.re.kr.
authors.forename: Kyung-Sook
authors.initials: KS
authors.lastname: Chung
mesh_terms: D000072283:A549 Cells; D000107:Acetylation; D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D000072224:Bcl-2-Like Protein 11 / Q000235:genetics / Q000378:metabolism; D002289:Carcinoma, Non-Small-Cell Lung / Q000188:drug therapy* / Q000201:enzymology / Q000235:genetics / Q000473:pathology; D004305:Dose-Response Relationship, Drug; D050881:E1A-Associated p300 Protein / Q000235:genetics / Q000378:metabolism*; D051766:Early Growth Response Protein 1 / Q000235:genetics / Q000378:metabolism*; D005260:Female; D000071316:Forkhead Box Protein O3 / Q000235:genetics / Q000378:metabolism*; D015972:Gene Expression Regulation, Neoplastic; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology*; D006801:Humans; D008175:Lung Neoplasms / Q000188:drug therapy* / Q000201:enzymology / Q000235:genetics / Q000473:pathology; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009285:Naphthoquinones / Q000494:pharmacology*; D051057:Proto-Oncogene Proteins c-akt / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference; D015398:Signal Transduction / Q000187:drug effects*; D056564:Sirtuin 1 / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D014162:Transfection; D023041:Xenograft Model Antitumor Assays
chemical_list: D000970:Antineoplastic Agents; C000606775:BCL2L11 protein, human; D000072224:Bcl-2-Like Protein 11; C497325:EGR1 protein, human; D051766:Early Growth Response Protein 1; C499517:FOXO3 protein, human; D000071316:Forkhead Box Protein O3; D056572:Histone Deacetylase Inhibitors; D009285:Naphthoquinones; C016101:shikonin; D050881:E1A-Associated p300 Protein; C494360:EP300 protein, human; D051057:Proto-Oncogene Proteins c-akt; C447939:SIRT1 protein, human; D056564:Sirtuin 1
keywords: AKT; EGR1; FOXO3a; SIRT1; Shikonin-induced apoptosis; p300