Paper - WANFANG MED ONLINE

You Position: Home > Paper

Epigenetic Silencing of Eyes Absent 4 Gene by Acute Myeloid Leukemia 1-Eight-twenty-one Oncoprotein Contributes to Leukemogenesis in t(8;21) Acute Myeloid Leukemia

( views:42, downloads:12 )

Author:: No author available

Journal Title:: Chinese Medical Journal

Issue:: 11

DOI:: 10.4103/0366-6999.182838

Key Word:: Acute Myeloid Gene 1-Eight-twenty-one;Acute Myeloid Leukemia;Epigenetics;Eyes Absent 4

Abstract： Background:The acute myeloid leukemia 1 (AML 1)-eight-twenty-one (ETO) fusion protein generated by the t(8;21)(q22;q22) translocation is considered to display a crucial role in leukemogenesis in AML.By focusing on the anti-leukemia effects of eyes absent 4 (EYA4) gene on AML cells,we investigated the biologic and molecular mechanism associated with AML1-ETO expressed in t(8;21) AML.Methods:Qualitative polymerase chain reaction (PCR),quantitative reverse transcription PCR (RT-PCR),and Western blotting analysis were used to observe the mRNA and protein expression levels of EYA4 in cell lines.Different plasmids (including mutant plasmids) of dual luciferase reporter vector were built to study the binding status of AML1-ETO to the promoter region of EYA4.Chromatin immunoprecipitation assay was used to study the epigenetic silencing mechanism of EYA4.Bisulfite sequencing was applied to detect the methylation status in EYA4 promoter region.The influence of EYA4 gene in the cell proliferation,apoptosis,and cell clone-forming ability was detected by the technique of Cell Counting Kit-8,flow cytometry,and clonogenic assay.Results:EYA4 gene was hypermethylated in AML1-ETO+ patients and its expression was down-regulated by 6-fold in Kasumi-1 and SKNO-1 cells,compared to HL-60 and SKNO-1-siA/E cells,respectively.We demonstrated that AML1-ETO triggered the epigenetic silencing of EYA4 gene by binding at AML1-binding sites and recruiting histone deacetylase 1 and DNA methyltransferases.Enhanced EYA4 expression levels inhibited cellular proliferation and suppressed cell colony formation in AML1-ETO+ cell lines.We also found EYA4 transfection increased apoptosis ofKasumi-1 and SKNO-1 cells by 1.6-fold and 1.4-fold compared to negative control,respectively.Conclusions:Our study identified EYA4 gene as targets for AML1-ETO and indicated it as a novel tumor suppressor gene.In addition,we provided evidence that EYA4 gene might be a novel therapeutic target and a potential candidate for treating AML 1-ETO+ t (8;21) AML.

Reference

[1]Alessandro, Gardini,Matteo, Cesaroni,Lucilla, Luzi,etc.AML1/ETO oncoprotein is directed to AML1 binding regions and co-localizes with AML1 and HEB on its targets.[J].2008,4.
[2]David, Grimwade,Robert K, Hills,Anthony V, Moorman,etc.Refinement of cytogenetic classification in acute myeloid leukemia: determination of prognostic significance of rare recurring chromosomal abnormalities among 5876 younger adult patients treated in the United Kingdom Medical Research Council trials.[J].2010,116.
[3]Xue Li,Oghi, Kenneth A.,Jie Zhang,etc.Eya protein phosphatase activity regulates Six1-Dach-Eya transcriptional effects in mammalian organogenesis.[J].2003,426(6964).
[4]Jessica, Nordlund,Lili, Milani,Anders, Lundmark,etc.DNA methylation analysis of bone marrow cells at diagnosis of acute lymphoblastic leukemia and at remission.[J].2012,7(4).
[5]Zardo G,Starnes LM,Racanicchi S,etc.Epigenetic silencing of the myelopoiesis regulator microRNA-223 by the AML1/ETO oncoprotein.[J].2007,12(5).
[6]Qumsiyeh M,Carroll A,Edwards C,etc.Patients with t(8; 21)(q22; q22) and acute myeloid leukemia have superior failure-free and overall survival when repetitive cycles of high-dose cytarabine are administered.[J].1999,17(12).
[7]Francesco, Fazi,Giuseppe, Zardo,Vania, Gelmetti,etc.Heterochromatic gene repression of the retinoic acid pathway in acute myeloid leukemia.[J].2007,109.
[8]V. S. Prassolov,A. V. Rulina,P. V. Spirin.Activated Leukemic Oncogenes AML1-ETO and c-kit: Role in Development of Acute Myeloid Leukemia and Current Approaches for Their Inhibition[J].2010,75(13).
[9]Rebay I,Jemc J.The eyes absent family of phosphotyrosine phosphatases: properties and roles in developmental regulation of transcription.[J].2007,76(0).
[10]Bennett, John M.,Catovsky, Daniel,Daniel, Marie T.,etc.Proposed Revised Criteria for the Classification of Acute Myeloid Leukemia.[J].1985,103(4).
[11]N, Martinez Soria,R, Tussiwand,P, Ziegler,etc.Transient depletion of RUNX1/RUNX1T1 by RNA interference delays tumour formation in vivo.[J].2009,23(1).
[12]Schonberger J,Wang L,Kim JB,etc.Mutation in the transcriptional coactivator EYA4 causes dilated cardiomyopathy and sensorineural hearing loss[J].2005,37(4).
[13]Felicetto, Ferrara,Luigi, Del Vecchio.Acute myeloid leukemia with t(8; 21)/AML1/ETO: a distinct biological and clinical entity.[J].2002,87.
[14]Markus, Pfister,Tímea, Tóth,Holger, Thiele,etc.A 4-bp insertion in the eya-homologous region (eyaHR) of EYA4 causes hearing impairment in a Hungarian family linked to DFNA10.[J].2002,8.
[15]Milani L,Lundmark A,Kiialainen A,etc.DNA methylation for subtype classification and prediction of treatment outcome in patients with childhood acute lymphoblastic leukemia.[J].2010,115(6).
[16]Hongzhi, Zou,Julian R, Molina,Jonathan J, Harrington,etc.Aberrant methylation of secreted frizzled-related protein genes in esophageal adenocarcinoma and Barrett's esophagus.[J].2005,116.
[17]Serve H,Stocking C,Muller Tidow C,etc.AML1/ETO induces self-renewal in hematopoietic progenitor cells via the Groucho-related amino-terminal AES protein.[J].2011,117(16).
[18]Luke F, Peterson,Anita, Boyapati,Eun-Young, Ahn,etc.Acute myeloid leukemia with the 8q22; 21q22 translocation: secondary mutational events and alternative t(8; 21) transcripts.[J].2007,110.
[19]Dahl HH,Hildebrand MS,Gardner RJ,etc.A novel splice site mutation in EYA4 causes DFNA10 hearing loss.[J].2007,143(14).
[20]Xiaoning, Gao,Ji, Lin,Li, Gao,etc.High expression of c-kit mRNA predicts unfavorable outcome in adult patients with t(8; 21) acute myeloid leukemia.[J].2015,10.
[21]Rayapureddi, Jayanagendra P.,Kattamuri, Chandramohan,Steinmetz, Brian D.,etc.Eyes absent represents a class of protein tyrosine phosphatases.[J].2003,426(6964).
[22]IM Wilson,EA Vucic,KSS Enfield,etc.EYA4 is inactivated biallelically at a high frequency in sporadic lung cancer and is associated with familial lung cancer risk[J].2014,33(36).
[23]Willemze R,Harousseau JL,Blaise D,etc.Identical outcome after autologous or allogeneic genoidentical hematopoietic stem-cell transplantation in first remission of acute myelocytic leukemia carrying inversion 16 or t(8; 21): a retrospective study from the european cooperative group for Blood a[J].2008,26(19).
[24]Lubbert M,Yalcin A,Duque Afonso J,etc.The HDAC class I-specific inhibitor entinostat (MS-275) effectively relieves epigenetic silencing of the LAT2 gene mediated by AML1/ETO.[J].2011,30(27).
[25]H, Narimatsu,T, Yokozawa,H, Iida,etc.Clinical characteristics and outcomes in patients with t(8;21) acute myeloid leukemia in Japan.[J].2008,22.
[26]Tina L, Tootle,Serena J, Silver,Erin L, Davies,etc.The transcription factor Eyes absent is a protein tyrosine phosphatase.[J].2003,426(6964).
[27]Klatt KK,Ahlquist DA,Osborn NK,etc.Aberrant methylation of the eyes absent 4 gene in ulcerative colitis-associated dysplasia.[J].2006,4(2).
[28]G, Marcucci,M A, Caligiuri,C D, Bloomfield.Molecular and clinical advances in core binding factor primary acute myeloid leukemia: a paradigm for translational research in malignant hematology.[J].2000,18.
[29]Min, Byung-Hoon,Tae, Chung Hyun,Rhee, Poong-Lyul,etc.EYA4 Acts as a New Tumor Suppressor Gene in Colorectal Cancer[J].2015,54(12).
[30]Zhuang,W.-Y.,Cen,J.-N.,Zhao,Y.,etc.Epigenetic silencing of Bcl-2, CEBPA and p14ARF by the AML1-ETO oncoprotein contributing to growth arrest and differentiation block in the U937 cell line[J].2013,30(1).
[31]Smith RJ,Wayne S,Van-Camp G,etc.Mutations in the transcriptional activator EYA4 cause late-onset deafness at the DFNA10 locus.[J].2001,10(3).
[32]Xun Hou.DNA Methylation Profiling Identifies EYA4 Gene as a Prognostic Molecular Marker in Hepatocellular Carcinoma[J].2014,21(12).
[33]P X, Xu,I, Woo,H, Her,etc.Mouse Eya homologues of the Drosophila eyes absent gene require Pax6 for expression in lens and nasal placode.[J].1997,124.
[34]Lin P,Chen L,Luthra R,etc.Acute myeloid leukemia harboring t(8;21)(q22;q22): a heterogeneous disease with poor outcome in a subset of patients unrelated to secondary cytogenetic aberrations.[J].2008,21(8).