MiR-29b suppresses the proliferation and migration of osteosarcoma cells by targeting CDK6

Abstract

Osteosarcoma is the most common primary sarcoma of bone, and it is a leading cause of cancer death among adolescents and young adults. However, the molecular mechanism underlying osteosarcoma carcinogenesis remains poorly understood. Recently, cyclin-dependent kinase 6 (CDK6) was identified as an important onco- gene. We found that CDK6 protein level, rather than CDK6 mRNA level, is much higher in osteosarcoma tissues than in normal adjacent tissues, which indicates a post-transcriptional mechanism involved in CDK6 regulation in osteosarcoma. MiRNAs are small non- coding RNAs that repress gene expression at the post- transcriptional level and have widely been shown to play important roles in many human cancers. In this study, we investigated the role of miR-29b as a novel regulator of CDK6 using bioinformatics methods. We demon- strated that CDK6 can be downregulated by miR-29b via binding to the 3′-UTR region in osteosarcoma cells. Furthermore, we identified an inverse correlation between miR-29b and CDK6 protein levels in osteosar- coma tissues. Finally, we examined the function of miR- 29b-driven repression of CDK6 expression in osteosarcoma cells. The results revealed that miR-29b acts as a tumor suppressor of osteosarcoma by target- ing CDK6 in the proliferation and migration processes. Taken together, our results highlight an important role for miR-29b in the regulation of CDK6 in osteosarcoma and may open new avenues for future osteosarcoma therapies.

Keywords

miR-29b / osteosarcoma / proliferation / migration / tumorigenesis

References

[1] Admassi D (2009) Osteosarcoma of medial cuniform bone. Ethiop Med J 47:305–308

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

[3] Broadhead ML, Clark JCM, Myers DE, Dass CR, Choong PFM (2011) The molecular pathogenesis of osteosarcoma: a review. Sarcoma. Article No.: 959248

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

[5] Carrington JC, Ambros V (2003) Role of microRNAs in plant and animal development. Science 301:336–338

[6] Chen X, Guo X, Zhang H, Xiang Y, Chen J, Yin Y, Cai X, Wang K, Wang G, Ba Y (2009) Role of miR-143 targeting KRAS in colorectal tumorigenesis. Oncogene 28:1385–1392

[7] Cortez MA, Nicoloso MS, Shimizu M, Rossi S, Gopisetty G, Molina JR, Carlotti C Jr, Tirapelli D, Neder L, Brassesco MS (2010) miR-29b and miR-125a regulate podoplanin and suppress invasion in glioblastoma. Genes Chromosom Cancer 49:981–990

[8] Dai N, Zhong ZY, Cun YP, Qing Y, Chen C, Jiang P, Li MX, Wang D (2013) Alteration of the microRNA expression profile in human osteosarcoma cells transfected with APE1 siRNA. Neoplasma 60:384–394

[9] Ekholm SV, Reed SI (2000) Regulation of G(1) cyclin-dependent kinases in the mammalian cell cycle. Curr Opin Cell Biol 12:676–684

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

[11] Fang JH, Zhou HC, Zeng CX, Yang J, Liu YL, Huang XZ, Zhang JP, Guan XY, Zhuang SM (2011) MicroRNA-29b suppresses tumor angiogenesis, invasion, and metastasis by regulating matrix metalloproteinase 2 expression. Hepatology 54:1729–1740

[12] Finn RS, Dering J, Conklin D, Kalous O, Cohen DJ, Desai AJ, Ginther C, Atefi M, Chen I, Fowst C (2009) PD 0332991, a selective cyclin D kinase 4/6 inhibitor, preferentially inhibits proliferation of luminal estrogen receptor-positive human breast cancer cell lines in vitro. Breast Cancer Res 11:R77

[13] Fry DW, Bedford DC, Harvey PH, Fritsch A, Keller PR, Wu ZP, Dobrusin E, Leopold WR, Fattaey A, Garrett MD (2001) Cell cycle and biochemical effects of PD 0183812—a potent inhibitor of the cyclin D-dependent kinases CDK4 and CDK6. J Biol Chem 276:16617–16623

[14] Fry DW, Harvey PJ, Keller PR, Elliott WL, Meade MA, Trachet E, Albassam M, Zheng XX, Leopold WR, Pryer NK (2004) Specific inhibition of cyclin-dependent kinase 4/6 by PD 0332991 and associated antitumor activity in human tumor xenografts. Mol Cancer Ther 3:1427–1437

[15] Garzon R, Heaphy CEA, Havelange V, Fabbri M, Volinia S, Tsao T, Zanesi N, Kornblau SM, Marcucci G, Calin GA (2009) MicroRNA 29b functions in acute myeloid leukemia. Blood 114:5331–5341

[16] Gorlick R (2009) Current concepts on the molecular biology of osteosarcoma. In: Jaffe N, Bruland OS, Bielack SS (eds) Pediatric and adolescent osteosarcoma. Springer, New York, pp 467–478

[17] Iorio MV, Croce CM (2012) MicroRNA dysregulation in cancer: diagnostics, monitoring and therapeutics. A comprehensive review. EMBO Mol Med 4:143–159

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

[19] Jones KB, Salah Z, Del Mare S, Galasso M, Gaudio E, Nuovo GJ, Lovat F, LeBlanc K, Palatini J, Randall RL (2012) miRNA signatures associate with pathogenesis and progression of osteosarcoma. Cancer Res 72:1865–1877

[20] Kansara M, Thomas DM (2007) Molecular pathogenesis of osteosarcoma. DNA Cell Biol 26:1–18

[21] Kole AJ, Swahari V, Hammond SM, Deshmukh M (2011) miR-29b is activated during neuronal maturation and targets BH3-only genes to restrict apoptosis. Genes Dev 25:125–130

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

[23] Kumar MS, Erkeland SJ, Pester RE, Chen CY, Ebert MS, Sharp PA, Jacks T (2008) Suppression of non-small cell lung tumor development by the let-7 microRNA family. Proc Natl Acad Sci USA 105:3903–3908

[24] Lee YS, Nakahara K, Pham JW, Kim K, He Z, Sontheimer EJ, Carthew RW (2004) Distinct roles for Drosophila Dicer-1 and Dicer-2 in the siRNA/miRNA silencing pathways. Cell 117:69–81

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

[26] Longhi A, Errani C, De Paolis M, Mercuri M, Bacci G (2006) Primary bone osteosarcoma in the pediatric age: state of the art. Cancer Treat Rev 32:423–436

[27] Ma L, Weinberg RA (2008) Micromanagers of malignancy: role of microRNAs in regulating metastasis. Trends Genet 24:448–456

[28] Malumbres M, Barbacid M (2001) To cycle or not to cycle: a critical decision in cancer. Nat Rev Cancer 1:222–231

[29] Malumbres M, Barbacid M (2009) Cell cycle, CDKs and cancer: a changing paradigm. Nat Rev Cancer 9:153–166

[30] Marina N, Gebhardt M, Teot L, Gorlick R (2004) Biology and therapeutic advances for pediatric osteosarcoma. Oncologist 9:422–441

[31] Matushansky I, Radparvar F, Skoultchi AI (2003) CDK6 blocks differentiation: coupling cell proliferation to the block to differen-tiation in leukemic cells. Oncogene 22:4143–4149

[32] Mott JL, Kobayashi S, Bronk SF, Gores GJ (2007) mir-29 regulates Mcl-1 protein expression and apoptosis. Oncogene 26:6133–6140

[33] Mott JL, Kurita S, Cazanave SC, Bronk SF, Werneburg NW, Fernandez-Zapico ME (2010) Transcriptional suppression of mir-29b-1/mir-29a Promoter by c-Myc, Hedgehog, and NF-kappaB. J Cell Biochem 110:1155–1164

[34] Musgrove EA, Caldon CE, Barraclough J, Stone A, Sutherland RL (2011) Cyclin D as a therapeutic target in cancer. Nat Rev Cancer 11:558–572

[35] Nicoloso MS, Spizzo R, Shimizu M, Rossi S, Calin GA (2009) MicroRNAs—the micro steering wheel of tumour metastases. Nat Rev Cancer 9:293–302

[36] Ottaviani G, Jaffe N (2009) The epidemiology of osteosarcoma. In: Jaffe N, Bruland OS, Bielack SS (eds) Pediatric and adolescent osteosarcoma. Springer, New York, pp 3–13

[37] Rossi M, Pitari MR, Amodio N, Di Martino MT, Conforti F, Leone E, Botta C, Paolino FM, Del Giudice T, Iuliano E (2013) miR-29b negatively regulates human osteoclastic cell differentiation and function: implications for the treatment of multiple myeloma-related bone disease. J Cell Physiol 228:1506–1515

[38] Sampath D, Liu C, Vasan K, Sulda M, Puduvalli VK, Wierda WG, Keating MJ (2012) Histone deacetylases mediate the silencing of miR-15a, miR-16, and miR-29b in chronic lymphocytic leukemia. Blood 119:1162–1172

[39] Wang C, Bian Z, Wei D, Zhang JG (2011) miR-29b regulates migration of human breast cancer cells. Mol Cell Biochem 352:197–207

[40] Weinstein IB, Joe AK (2006) Mechanisms of disease: oncogene addiction—a rationale for molecular targeting in cancer therapy. Nat Clin Pract Oncol 3:448–457

[41] Zhang KX, Zhang CY, Liu L, Zhou JH (2014) A key role of microRNA-29b in suppression of osteosarcoma cell proliferation and migration via modulation of VEGF. Int J Clin Exp Pathol 7:5701–5708

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