Abstract
MicroRNAs (miRNAs) are short non coding RNAs that regulate the gene expression and play a relevant role in physiopathological mechanisms such as development, proliferation, death, and differentiation of normal and cancer cells. Recently, abnormal expression of miRNAs has been reported in most of solid or hematopoietic malignancies, including multiple myeloma (MM), where miRNAs have been found deeply dysregulated and act as oncogenes or tumor suppressors. Presently, the most recognized approach for definition of miRNA portraits is based on microarray profiling analysis. We here describe a workflow based on the identification of dysregulated miRNAs in plasma cells from MM patients based on Affymetrix technology. We describe how it is possible to search miRNA putative targets performing whole gene expression profile on MM cell lines transfected with miRNA mimics or inhibitors followed by luciferase reporter assay to analyze the specific targeting of the 3′untranslated region (UTR) sequence of a mRNA by selected miRNAs. These technological approaches are suitable strategies for the identification of relevant druggable targets in MM.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Anderson KC (2014) Multiple myeloma. Hematol Oncol Clin North Am 28:xi–xii. doi:10.1016/j.hoc.2014.08.001
Tagliaferri P et al (2012) Promises and challenges of microRNA-based treatment of multiple myeloma. Current Cancer Drug Targets 12:838–846
Tassone P, Tagliaferri P (2012) Editorial: new approaches in the treatment of multiple myeloma: from target-based agents to the new era of microRNAs (dedicated to the memory of Prof. Salvatore Venuta). Curr Cancer Drug Targets 12:741–742
Rossi M et al (2013) From target therapy to miRNA therapeutics of human multiple myeloma: theoretical and technological issues in the evolving scenario. Curr Drug Targets 14:1144–1149
Rossi M et al (2014) MicroRNA and multiple myeloma: from laboratory findings to translational therapeutic approaches. Curr Pharm Biotechnol 15:459–467
Misso G et al (2013) Emerging pathways as individualized therapeutic target of multiple myeloma. Expert Opin Biol Ther 13(Suppl 1):S95–S109. doi:10.1517/14712598.2013.807338
Misso G et al (2014) Mir-34: a new weapon against cancer? Mol Ther Nucleic Acids 3:e194. doi:10.1038/mtna.2014.47
Lionetti M et al (2013) Biological and clinical relevance of miRNA expression signatures in primary plasma cell leukemia. Clin Cancer Res 19:3130–3142. doi:10.1158/1078-0432.CCR-12-2043
Lionetti M, Agnelli L, Lombardi L, Tassone P, Neri A (2012) MicroRNAs in the pathobiology of multiple myeloma. Curr Cancer Drug Targets 12:823–837
Amodio N et al (2013) miR-29b induces SOCS-1 expression by promoter demethylation and negatively regulates migration of multiple myeloma and endothelial cells. Cell Cycle 12:3650–3662. doi:10.4161/cc.26585
Amodio N, Di Martino MT, Neri A, Tagliaferri P, Tassone P (2013) Non-coding RNA: a novel opportunity for the personalized treatment of multiple myeloma. Expert Opin Biol Ther 13(Suppl 1):S125–S137. doi:10.1517/14712598.2013.796356
Amodio N et al (2012) DNA-demethylating and anti-tumor activity of synthetic miR-29b mimics in multiple myeloma. Oncotarget 3:1246–1258
Di Martino MT et al (2014) In vivo activity of miR-34a mimics delivered by stable nucleic acid lipid particles (SNALPs) against multiple myeloma. PloS One 9:e90005. doi:10.1371/journal.pone.0090005
Di Martino MT et al (2014) In vitro and in vivo activity of a novel locked nucleic acid (LNA)-inhibitor-miR-221 against multiple myeloma cells. PloS One 9:e89659. doi:10.1371/journal.pone.0089659
Di Martino MT et al (2012) Synthetic miR-34a mimics as a novel therapeutic agent for multiple myeloma: in vitro and in vivo evidence. Clin Cancer Res 18:6260–6270. doi:10.1158/1078-0432.CCR-12-1708
Leone E et al (2013) Targeting miR-21 inhibits in vitro and in vivo multiple myeloma cell growth. Clin Cancer Res 19:2096–2106. doi:10.1158/1078-0432.CCR-12-3325
Leotta M et al (2014) A p53-dependent tumor suppressor network is induced by selective miR-125a-5p inhibition in multiple myeloma cells. J Cell Physiol 229:2106–2116. doi:10.1002/jcp.24669
Rossi M et al (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. doi:10.1002/jcp.24306
Scognamiglio I et al (2014) Transferrin-conjugated SNALPs encapsulating 2′-O-methylated miR-34a for the treatment of multiple myeloma. Biomed Res Int 2014:217365. doi:10.1155/2014/217365
Monroig PD, Chen L, Zhang S, Calin GA (2014) Small molecule compounds targeting miRNAs for cancer therapy. Adv Drug Deliv Rev. doi:10.1016/j.addr.2014.09.002
Amodio N et al (2012) miR-29b sensitizes multiple myeloma cells to bortezomib-induced apoptosis through the activation of a feedback loop with the transcription factor Sp1. Cell Death Dis 3:e436. doi:10.1038/cddis.2012.175
Di Martino MT et al (2013) In vitro and in vivo anti-tumor activity of miR-221/222 inhibitors in multiple myeloma. Oncotarget 4:242–255
Lionetti M et al (2009) Identification of microRNA expression patterns and definition of a microRNA/mRNA regulatory network in distinct molecular groups of multiple myeloma. Blood 114:e20–e26. doi:10.1182/blood-2009-08-237495
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 25:402–408. doi:10.1006/meth.2001.1262
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2015 Springer Science+Business Media New York
About this protocol
Cite this protocol
Di Martino, M.T., Amodio, N., Tassone, P., Tagliaferri, P. (2015). Functional Analysis of microRNA in Multiple Myeloma. In: Guzzi, P. (eds) Microarray Data Analysis. Methods in Molecular Biology, vol 1375. Humana Press, New York, NY. https://doi.org/10.1007/7651_2015_250
Download citation
DOI: https://doi.org/10.1007/7651_2015_250
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3172-9
Online ISBN: 978-1-4939-3173-6
eBook Packages: Springer Protocols