miR-10a inhibits cell proliferation and promotes cell apoptosis by targeting BCL6 in diffuse large B-cell lymphoma

Abstract

The BCL6 (B-Cell Lymphoma 6) gene is a proto-oncogene that is often expressed in diffuse large B-cell lymphomas (DLBCLs). BCL6 loss of function can kill DLBCL cells, demonstrating that BCL6 is necessary for the survival of DLBCL cells and could be a therapeutic target. In this study, we found that BCL6 protein levels were consistently upregulated in DLBCL tissues, whereas its mRNA levels varied randomly in tissues, suggesting that a post-transcriptional mechanism was involved in BCL6 regulation. We used bioinformatics analysis to search for miRNAs, which potentially target BCL6, and identified specific targeting sites for miR-10a in the 3′-untranslated region (3′-UTR) of BCL6. We further identified an inverse correlation between miR-10a levels and BCL6 protein levels, but not mRNA levels, in DLBCL tumor tissue samples. By overexpressing or knocking down miR-10a in DLBCL cells, we experimentally validated that miR-10a directly recognizes the 3′-UTR of the BCL6 transcript and regulated BCL6 expression. Furthermore, we demonstrated that negatively regulating BCL6 by miR-10a suppressed the proliferation and promoted apoptosis of DLBCL cells.

Keywords

microRNA / miR-10a / BCL6 / DLBCL / proliferation / apoptosis

References

[1] Agirre X, Jimenez-Velasco A, San Jose-Eneriz E, Garate L, Bandres E, Cordeu L, Aparicio O, Saez B, Navarro G, Vilas-Zornoza A, Perez-Roger I, Garcia-Foncillas J, Torres A, Heiniger A, Calasanz MJ, Fortes P, Roman-Gomez J, Prosper F (2008) Down-regulation of hsa-miR-10a in chronic myeloid leukemia CD34+ cells increases USF2-mediated cell growth. Mol Cancer Res 6 (12):1830–1840

[2] Ambros V (2004) The functions of animal microRNAs. Nature 431 (7006):350–355

[3] Baron BW, Anastasi J, Montag A, Huo D, Baron RM, Karrison T, Thirman MJ, Subudhi SK, Chin RK, Felsher DW, Fu YX, McKeithan TW, Baron JM (2004) The human BCL6 transgene promotes the development of lymphomas in the mouse. Proc Natl Acad Sci USA 101(39):14198–14203

[4] Bartel DP (2004) MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116(2):281–297

[5] Basso K, Dalla-Favera R (2012) Roles of BCL6 in normal and transformed germinal center B cells. Immunol Rev 247(1):172–183

[6] Bertolo C, Roa S, Sagardoy A, Mena-Varas M, Robles EF, Martinez-Ferrandis JI, Sagaert X, Tousseyn T, Orta A, Lossos IS, Amar S, Natkunam Y, Briones J, Melnick A, Malumbres R, Martinez-Climent JA (2013) LITAF, a BCL6 target gene, regulates autophagy in mature B-cell lymphomas. Br J Haematol 162 (5):621–630

[7] Calin GA, Croce CM (2006) MicroRNA signatures in human cancers. Nat Rev Cancer 6(11):857–866

[8] Cerchietti L, Melnick A (2013) Targeting BCL6 in diffuse large B-cell lymphoma: what does this mean for the future treatment? Expert Rev Hematol 6(4):343–345

[9] Chen X, Guo X, Zhang H, Xiang Y, Chen J, Yin Y, Cai X, Wang K, Wang G, Ba Y, Zhu L, Wang J, Yang R, Zhang Y, Ren Z, Zen K, Zhang J, Zhang CY (2009) Role of miR-143 targeting KRAS in colorectal tumorigenesis. Oncogene 28(10):1385–1392

[10] Duan S, Cermak L, Pagan JK, Rossi M, Martinengo C, di Celle PF, Chapuy B, Shipp M, Chiarle R, Pagano M (2012) FBXO11 targets BCL6 for degradation and is inactivated in diffuse large B-cell lymphomas. Nature 481(7379):90–93

[11] Ding J, Dirks WG, Ehrentraut S, Geffers R, MacLeod RA, Nagel S, Pommerenke C, Romani J, Scherr M, Vaas LA, Zaborski M, Drexler HG, Quentmeier H (2015) BCL6—regulated by AhR/ARNT and wild-type MEF2B—drives expression of germinal center markers MYBL1 and LMO2. Haematologica 100(6):801–809

[12] Esquela-Kerscher A, Slack FJ (2006) Oncomirs—microRNAs with a role in cancer. Nat Rev Cancer 6(4):259–269

[13] Gao P, Li Q, Wang Z, Yan F, Lu C, Cao X (2014) Significance of BCL6, MYC, P53 genes abnormalities for the prognosis of diffuse large B-cell lymphoma. Zhonghua Yi Xue Yi Chuan Xue Za Zhi 31 (5):628–631

[14] Gaur A, Jewell DA, Liang Y, Ridzon D, Moore JH, Chen C, Ambros VR, Israel MA (2007) Characterization of microRNA expression levels and their biological correlates in human cancer cell lines. Cancer Res 67(6):2456–2468

[15] Gerrard M, Waxman IM, Sposto R, Auperin A, Perkins SL, Goldman S, Harrison L, Pinkerton R, McCarthy K, Raphael M, Patte C, Cairo MS (2013) Outcome and pathologic classification of children and adolescents with mediastinal large B-cell lymphoma treated with FAB/LMB96 mature B-NHL therapy. Blood 121 (2):278–285

[16] Hatzi K, Jiang Y, Huang C, Garrett-Bakelman F, Gearhart MD, Giannopoulou EG, Zumbo P, Kirouac K, Bhaskara S, Polo JM, Kormaksson M, MacKerell ADJr, Xue F, Mason CE, Hiebert SW, Prive GG, Cerchietti L, Bardwell VJ, Elemento O, Melnick A (2013) A hybrid mechanism of action for BCL6 in B cells defined by formation of functionally distinct complexes at enhancers and promoters. Cell Rep 4(3):578–588

[17] Havelange V, Ranganathan P, Geyer S, Nicolet D, Huang X, Yu X, Volinia S, Kornblau SM, Andreeff M, Croce CM, Marcucci G, Bloomfield CD, Garzon R (2014) Implications of the miR-10 family in chemotherapy response of NPM1-mutated AML. Blood 123(15):2412–2415

[18] He L, Hannon GJ (2004) MicroRNAs: small RNAs with a big role in gene regulation. Nat Rev Genet 5(7):522–531

[19] Hollister K, Kusam S, Wu H, Clegg N, Mondal A, Sawant DV, Dent AL (2013) Insights into the role of Bcl6 in follicular Th cells using a new conditional mutant mouse model. J Immunol 191(7):3705–3711

[20] Ivey KN, Srivastava D (2015) microRNAs as developmental regulators. Cold Spring Harb Perspect Biol 7(7):a008144

[21] John B, Enright AJ, Aravin A, Tuschl T, Sander C, Marks DS (2004) Human microRNA targets. PLoS Biol 2(11):e363

[22] Jung I, Aguiar RC (2009) microRNA-155 expression and outcome in diffuse large B-cell lymphoma. Br J Haematol 144(1):138–140

[23] Khan S, Wall D, Curran C, Newell J, Kerin MJ, Dwyer RM (2015) microRNA-10a is reduced in breast cancer and regulated in part through retinoic acid. BMC Cancer 15(1):345

[24] Koscianska E, Krzyzosiak WJ (2014) Current understanding of the role of microRNAs in spinocerebellar ataxias. Cerebellum Ataxias 1:7

[25] Krek A, Grun D, Poy MN, Wolf R, Rosenberg L, Epstein EJ, MacMenamin P, da Piedade I, Gunsalus KC, Stoffel M, Rajewsky N (2005) Combinatorial microRNA target predictions. Nat Genet 37(5):495–500

[26] Krol J, Loedige I, Filipowicz W (2010) The widespread regulation of microRNA biogenesis, function and decay. Nat Rev Genet 11 (9):597–610

[27] Lewis BP, Shih IH, Jones-Rhoades MW, Bartel DP, Burge CB (2003) Prediction of mammalian microRNA targets. Cell 115(7):787–798

[28] Parekh S, Polo JM, Shaknovich R, Juszczynski P, Lev P, Ranuncolo SM, Yin Y, Klein U, Cattoretti G, Dalla Favera R, Shipp MA, Melnick A (2007) BCL6 programs lymphoma cells for survival and differentiation through distinct biochemical mechanisms. Blood 110(6):2067–2074

[29] Parekh S, Prive G, Melnick A (2008) Therapeutic targeting of the BCL6 oncogene for diffuse large B-cell lymphomas. Leuk Lymphoma 49(5):874–882

[30] Pillai RS, Bhattacharyya SN, Filipowicz W (2007) Repression of protein synthesis by miRNAs: how many mechanisms? Trends Cell Biol 17(3):118–126

[31] Polo JM, Dell’Oso T, Ranuncolo SM, Cerchietti L, Beck D, Da Silva GF, Prive GG, Licht JD, Melnick A (2004) Specific peptide interference reveals BCL6 transcriptional and oncogenic mechanisms in B-cell lymphoma cells. Nat Med 10(12):1329–1335

[32] Roehle A, Hoefig KP, Repsilber D, Thorns C, Ziepert M, Wesche KO, Thiere M, Loeffler M, Klapper W, Pfreundschuh M, Matolcsy A, Bernd HW, Reiniger L, Merz H, Feller AC (2008) microRNA signatures characterize diffuse large B-cell lymphomas and follicular lymphomas. Br J Haematol 142(5):732–744

[33] Saito Y, Liang G, Egger G, Friedman JM, Chuang JC, Coetzee GA, Jones PA (2006) Specific activation of microRNA-127 with downregulation of the proto-oncogene BCL6 by chromatinmodifying drugs in human cancer cells. Cancer Cell 9(6):435–443

[34] Shaffer AL, Yu X, He Y, Boldrick J, Chan EP, Staudt LM (2000) BCL-6 represses genes that function in lymphocyte differentiation, inflammation, and cell cycle control. Immunity 13(2):199–212

[35] Takahashi H, Kanno T, Nakayamada S, Hirahara K, Sciume G, Muljo SA, Kuchen S, Casellas R, Wei L, Kanno Y, O’Shea JJ (2012) TGF-beta and retinoic acid induce the microRNA miR-10a, which targets Bcl-6 and constrains the plasticity of helper T cells. Nat Immunol 13(6):587–595

[36] Van Den Neste E, Schmitz N, Mounier N, Gill D, Linch D, Trneny M, Milpied N, Radford J, Ketterer N, Shpilberg O, Duhrsen U, Ma D, Briere J, Thieblemont C, Salles G, Moskowitz CH, Glass B, Gisselbrecht C (2015) Outcome of patients with relapsed diffuse large B-cell lymphoma who fail second-line salvage regimens in the International CORAL study. Bone Marrow Transplant. 51 (1):51–57

[37] Wang K, Xu Z, Wang N, Xu T, Zhu M (2014) microRNA and gene networks in human diffuse large B-cell lymphoma. Oncol Lett 8 (5):2225–2232

[38] Westin JR, Fayad LE (2009) Beyond R-CHOP and the IPI in largecell lymphoma: molecular markers as an opportunity for stratification. Curr Hematol Malig Rep 4(4):218–224

[39] Winter JN, Weller EA, Horning SJ, Krajewska M, Variakojis D, Habermann TM, Fisher RI, Kurtin PJ, Macon WR, Chhanabhai M, Felgar RE, Hsi ED, Medeiros LJ, Weick JK, Reed JC, Gascoyne RD (2006) Prognostic significance of Bcl-6 protein expression in DLBCL treated with CHOP or R-CHOP: a prospective correlative study. Blood 107(11):4207–4213

[40] Yan Y, Luo YC, Wan HY, Wang J, Zhang PP, Liu M, Li X, Li S, Tang H (2013) microRNA-10a is involved in the metastatic process by regulating Eph tyrosine kinase receptor A4-mediated epithelialmesenchymal transition and adhesion in hepatoma cells. Hepatology 57(2):667–677

[41] Yatomi Y (2012) From FAB classification to WHO classification of tumors of hematopoietic and lymphoid tissue. Rinsho Byori 60 (6):550–552

[42] Yu D, Rao S, Tsai LM, Lee SK, He Y, Sutcliffe EL, Srivastava M, Linterman M, Zheng L, Simpson N, Ellyard JI, Parish IA, Ma CS, Li QJ, Parish CR, Mackay CR, Vinuesa CG (2009) The transcriptional repressor Bcl-6 directs T follicular helper cell lineage commitment. Immunity 31(3):457–468

[43] Zeng T, Li G (2014) microRNA10a enhances the metastatic potential of cervical cancer cells by targeting phosphatase and tensin homologue. Mol Med Rep 10(3):1377–1382

[44] Zhang L, Huang J, Yang N, Greshock J, Megraw MS, Giannakakis A, Liang S, Naylor TL, Barchetti A, Ward MR, Yao G, Medina A, O’Brien-Jenkins A, Katsaros D, Hatzigeorgiou A, Gimotty PA, Weber BL, Coukos G (2006) microRNAs exhibit high frequency genomic alterations in human cancer. Proc Natl Acad Sci USA 103(24):9136–9141

RIGHTS & PERMISSIONS

2016 The Author(s) 2016. This article is published with open access at Springerlink.com and journal.hep.com.cn