Luciano Mazzoccoli1, Marcela Cristina Robaina1, Alexandre Gustavo Apa2, Martin Bonamino3,4, Luciana Wernersbach Pinto5, Eduardo Queiroga6, Carlos E Bacchi6, Claudete Esteves Klumb7,8. 1. Programa de Pesquisa em Hemato-Oncologia Molecular, Instituto Nacional de Câncer, Rio de Janeiro, Brazil. 2. Serviço de Hematologia, Instituto Nacional de Câncer, Rio de Janeiro, Brazil. 3. Programa de Carcinogênese Molecular, Instituto Nacional de Câncer, Rio de Janeiro, Brazil. 4. Fundação Instituto Oswaldo Cruz, Vice-presidência de Pesquisa e Laboratórios de Referência, Rio de Janeiro, Brazil. 5. Divisão de Patologia, Instituto Nacional de Câncer, Rio de Janeiro, Brazil. 6. Laboratório Bacchi, Consultoria em Patologia, Botucatu, São Paulo, Brazil. 7. Programa de Pesquisa em Hemato-Oncologia Molecular, Instituto Nacional de Câncer, Rio de Janeiro, Brazil. cklumb@inca.gov.br. 8. Laboratório de Hemato-Oncologia Celular e Molecular, Instituto Nacional de Câncer, Praça da Cruz Vermelha, 23, 6° andar, ala C, Rio de Janeiro, RJ, CEP: 20230-130, Brazil. cklumb@inca.gov.br.
Abstract
PURPOSE: Burkitt lymphoma (BL) is a B-cell lymphoma frequently diagnosed in children. It is characterized by MYC translocations, which lead to the constitutive expression of the MYC oncogene. MYC contributes to miR-29 repression through an E-box MYC binding site on the miR-29b-1/miR-29a promoter region. We evaluated the role of miR-29a/b/c and their predicted targets in BL pathogenesis. METHODS: Mature sequences of miR-29a/b/c were transfected to the BL cell lines BL41 and Raji, and evaluated for DNMT3B, MCL1, BIM, CDK6, AKT and TCL1 protein expression as well as for MCL-1 and CDK6 mRNA expression. BL cells were treated with 5-aza-2'-deoxycytidine (decitabine) and evaluated for miR29 expressions and methylation status. DNMT3B inhibition was performed by DNMT3B siRNA. RESULTS: Ectopic expression of miR-29s in BL cells decreased CDK6, DNMT3B, TCL1 and MCL-1 protein levels, but CDK6 and MCL-1 mRNA expression was unaffected by miR-29. Decitabine enhanced miR-29 expression levels and decreased CDK6 protein expression. Additionally, inhibition of DNMT3B by siRNA increased miR-29a/b expression. Notably, the miR-29a/b1 and miR-29b2/c promoter genes showed methylated CpG sequences that were demethylated after decitabine treatments. Furthermore, MYC-negative tumours had higher levels of miR-29 expression compared with MYC-translocated cases, suggesting that MYC regulates miR-29 in BL tumours. CONCLUSIONS: Our results suggest a significant role for miR-29s in BL pathogenesis in altering the expression of targets involved in critical cancer pathways, such as cell cycle control, apoptosis inhibition and DNA methylation. Moreover, methylation-mediated miR-29 epigenetic silencing may occur during BL development.
PURPOSE:Burkitt lymphoma (BL) is a B-cell lymphoma frequently diagnosed in children. It is characterized by MYC translocations, which lead to the constitutive expression of the MYC oncogene. MYC contributes to miR-29 repression through an E-box MYC binding site on the miR-29b-1/miR-29a promoter region. We evaluated the role of miR-29a/b/c and their predicted targets in BL pathogenesis. METHODS: Mature sequences of miR-29a/b/c were transfected to the BL cell lines BL41 and Raji, and evaluated for DNMT3B, MCL1, BIM, CDK6, AKT and TCL1 protein expression as well as for MCL-1 and CDK6 mRNA expression. BL cells were treated with 5-aza-2'-deoxycytidine (decitabine) and evaluated for miR29 expressions and methylation status. DNMT3B inhibition was performed by DNMT3B siRNA. RESULTS: Ectopic expression of miR-29s in BL cells decreased CDK6, DNMT3B, TCL1 and MCL-1 protein levels, but CDK6 and MCL-1 mRNA expression was unaffected by miR-29. Decitabine enhanced miR-29 expression levels and decreased CDK6 protein expression. Additionally, inhibition of DNMT3B by siRNA increased miR-29a/b expression. Notably, the miR-29a/b1 and miR-29b2/c promoter genes showed methylated CpG sequences that were demethylated after decitabine treatments. Furthermore, MYC-negative tumours had higher levels of miR-29 expression compared with MYC-translocated cases, suggesting that MYC regulates miR-29 in BL tumours. CONCLUSIONS: Our results suggest a significant role for miR-29s in BL pathogenesis in altering the expression of targets involved in critical cancer pathways, such as cell cycle control, apoptosis inhibition and DNA methylation. Moreover, methylation-mediated miR-29 epigenetic silencing may occur during BL development.
Authors: Michael A Teitell; Mark A Lones; Sherrie L Perkins; Warren G Sanger; Mitchell S Cairo; Jonathan W Said Journal: Am J Clin Pathol Date: 2005-10 Impact factor: 2.493
Authors: Eugenia Haralambieva; Ed Schuuring; Stefano Rosati; Carel van Noesel; Patty Jansen; Inge Appel; Jeroen Guikema; Henry Wabinga; Luiz Fernando Bleggi-Torres; King Lam; Eva van den Berg; Clemens Mellink; Shama van Zelderen-Bhola; Philip Kluin Journal: Genes Chromosomes Cancer Date: 2004-05 Impact factor: 5.006
Authors: Pablo Landgraf; Mirabela Rusu; Robert Sheridan; Alain Sewer; Nicola Iovino; Alexei Aravin; Sébastien Pfeffer; Amanda Rice; Alice O Kamphorst; Markus Landthaler; Carolina Lin; Nicholas D Socci; Leandro Hermida; Valerio Fulci; Sabina Chiaretti; Robin Foà; Julia Schliwka; Uta Fuchs; Astrid Novosel; Roman-Ulrich Müller; Bernhard Schermer; Ute Bissels; Jason Inman; Quang Phan; Minchen Chien; David B Weir; Ruchi Choksi; Gabriella De Vita; Daniela Frezzetti; Hans-Ingo Trompeter; Veit Hornung; Grace Teng; Gunther Hartmann; Miklos Palkovits; Roberto Di Lauro; Peter Wernet; Giuseppe Macino; Charles E Rogler; James W Nagle; Jingyue Ju; F Nina Papavasiliou; Thomas Benzing; Peter Lichter; Wayne Tam; Michael J Brownstein; Andreas Bosio; Arndt Borkhardt; James J Russo; Chris Sander; Mihaela Zavolan; Thomas Tuschl Journal: Cell Date: 2007-06-29 Impact factor: 41.582
Authors: M Hashimoto; F Suizu; W Tokuyama; H Noguchi; N Hirata; M Matsuda-Lennikov; T Edamura; M Masuzawa; N Gotoh; S Tanaka; M Noguchi Journal: Oncogenesis Date: 2013-09-16 Impact factor: 7.485
Authors: Michelle Van Sinderen; Meaghan Griffiths; Ellen Menkhorst; Keith Niven; Evdokia Dimitriadis Journal: Oncol Lett Date: 2019-07-09 Impact factor: 2.967
Authors: Shatovisha Dey; Jason J Kwon; Sheng Liu; Gabriel A Hodge; Solaema Taleb; Teresa A Zimmers; Jun Wan; Janaiah Kota Journal: Mol Cancer Res Date: 2019-10-29 Impact factor: 5.852
Authors: Mirco Di Marco; Alice Ramassone; Sara Pagotto; Eleni Anastasiadou; Angelo Veronese; Rosa Visone Journal: Int J Mol Sci Date: 2018-10-12 Impact factor: 5.923
Authors: Yun Zhao; Wei Huang; Tae-Min Kim; Yuchae Jung; Lata G Menon; Hongyan Xing; Hongwei Li; Rona S Carroll; Peter J Park; Hong Wei Yang; Mark D Johnson Journal: J Exp Clin Cancer Res Date: 2019-01-25