In Vivo Imaging of miR-221 Biogenesis in Papillary Thyroid Carcinoma (original) (raw)
Abstract
Purpose
To investigate the overexpression of miR-221 in papillary thyroid carcinoma (PTC), we developed a Gaussia luciferase (Gluc) system regulated by miR-221.
Procedures
Quantities of primary or mature miR-221 in normal thyroid cells (HT-ori3) and in PTC (NPA, TPC-1) were measured by quantitative real-time polymerase chain reaction. Cytomegalovirus (CMV)/Gluc-3xPT_miR221, which included three perfect complementary target sequences repeats of miR221 in the 3′-untranslated region of Gluc, was transfected into cells with pre-miR-221 or anti-miR-221 and Gluc activities were then compared in vitro and in vivo.
Results
Primary or mature miR-221 were overexpressed in PTC as compared with HT-ori3. In cells transfected with the Gaussia luciferase reporter system (CMV/Gluc-3xPT_miR221), Gluc activities were regulated according to miR-221 levels in vitro and in vivo.
Conclusions
These results suggest that the devised CMV/Gluc-3xPT_miR221 system may be a useful tool for monitoring quantities of endogenous miR-221 in cells or living organisms.
Access this article
Subscribe and save
- Get 10 units per month
- Download Article/Chapter or eBook
- 1 Unit = 1 Article or 1 Chapter
- Cancel anytime Subscribe now
Buy Now
Price excludes VAT (USA)
Tax calculation will be finalised during checkout.
Instant access to the full article PDF.
Similar content being viewed by others
References
- Lee RC, Feinbaum RL, Ambros V (1993) The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14. Cell 75:843–854
Article PubMed CAS Google Scholar - Lu J, Getz G, Miska EA et al (2005) MicroRNA expression profiles classify human cancers. Nature 435:834–838
Article PubMed CAS Google Scholar - Kloosterman WP, Plasterk RH (2006) The diverse functions of microRNAs in animal development and disease. Dev Cell 11:441–450
Article PubMed CAS Google Scholar - Reinhart BJ, Slack FJ, Basson M et al (2000) The 21-nucleotide let-7 RNA regulates developmental timing in Caenorhabditis elegans. Nature 403:901–906
Article PubMed CAS Google Scholar - Bushati N, Cohen SM (2007) microRNA functions. Annu Rev Cell Dev Biol 23:175–205
Article PubMed CAS Google Scholar - Kim VN (2005) MicroRNA biogenesis: coordinated cropping and dicing. Nat Rev Mol Cell Biol 6:376–385
Article PubMed CAS Google Scholar - Zeng Y, Yi R, Cullen BR (2003) MicroRNAs and small interfering RNAs can inhibit mRNA expression by similar mechanisms. Proc Natl Acad Sci U S A 100:9779–9784
Article PubMed CAS Google Scholar - Valencia-Sanchez MA, Liu J, Hannon GJ, Parker R (2006) Control of translation and mRNA degradation by miRNAs and siRNAs. Genes Dev 20:515–524
Article PubMed CAS Google Scholar - Obernosterer G, Leuschner PJ, Alenius M, Martinez J (2006) Post-transcriptional regulation of microRNA expression. Rna 12:1161–1167
Article PubMed CAS Google Scholar - Weiler J, Hunziker J, Hall J (2006) Anti-miRNA oligonucleotides (AMOs): ammunition to target miRNAs implicated in human disease. Gene Ther 13:496–502
Article PubMed CAS Google Scholar - Brennecke J, Hipfner DR, Stark A, Russell RB, Cohen SM (2003) bantam encodes a developmentally regulated microRNA that controls cell proliferation and regulates the proapoptotic gene hid in Drosophila. Cell 113:25–36
Article PubMed CAS Google Scholar - Osada H, Takahashi T (2007) MicroRNAs in biological processes and carcinogenesis. Carcinogenesis 28:2–12
Article PubMed CAS Google Scholar - Tsuchiya S, Okuno Y, Tsujimoto G (2006) MicroRNA: biogenetic and functional mechanisms and involvements in cell differentiation and cancer. J Pharmacol Sci 101:267–270
Article PubMed CAS Google Scholar - Zhang B, Pan X, Cobb GP, Anderson TA (2007) microRNAs as oncogenes and tumor suppressors. Dev Biol 302:1–12
Article PubMed CAS Google Scholar - Esquela-Kerscher A, Slack FJ (2006) Oncomirs—microRNAs with a role in cancer. Nat Rev Cancer 6:259–269
Article PubMed CAS Google Scholar - Pallante P, Visone R, Ferracin M et al (2006) MicroRNA deregulation in human thyroid papillary carcinomas. Endocr Relat Cancer 13:497–508
Article PubMed CAS Google Scholar - Felli N, Fontana L, Pelosi E et al (2005) MicroRNAs 221 and 222 inhibit normal erythropoiesis and erythroleukemic cell growth via kit receptor down-modulation. Proc Natl Acad Sci USA 102:18081–18086
Article PubMed CAS Google Scholar - He H, Jazdzewski K, Li W et al (2005) The role of microRNA genes in papillary thyroid carcinoma. Proc Natl Acad Sci U S A 102:19075–19080
Article PubMed CAS Google Scholar - Nikiforova MN, Tseng GC, Steward D, Diorio D, Nikiforov YE (2008) MicroRNA expression profiling of thyroid tumors: biological significance and diagnostic utility. J Clin Endocrinol Metab 93(5):1600–1608
Article PubMed CAS Google Scholar - Gillies JK, Lorimer IA (2007) Regulation of p27Kip1 by miRNA 221/222 in glioblastoma. Cell Cycle 6:2005–2009
PubMed CAS Google Scholar - Doubrovin M, Serganova I, Mayer-Kuckuk P, Ponomarev V, Blasberg RG (2004) Multimodality in vivo molecular-genetic imaging. Bioconjug Chem 15:1376–1388
Article PubMed CAS Google Scholar - Lee JY, Kim S, Hwang DW et al (2008) Development of a dual-luciferase reporter system for in vivo visualization of MicroRNA biogenesis and posttranscriptional regulation. J Nucl Med 49(2):285–294
Article PubMed CAS Google Scholar - Ottobrini L, Ciana P, Biserni A, Lucignani G, Maggi A (2006) Molecular imaging: a new way to study molecular processes in vivo. Mol Cell Endocrinol 246:69–75
Article PubMed CAS Google Scholar - Gould SJ, Subramani S (1988) Firefly luciferase as a tool in molecular and cell biology. Anal Biochem 175:5–13
Article PubMed CAS Google Scholar - Brasier AR, Tate JE, Habener JF (1989) Optimized use of the firefly luciferase assay as a reporter gene in mammalian cell lines. Biotechniques 7:1116–1122
PubMed CAS Google Scholar - Tannous BA, Kim DE, Fernandez JL, Weissleder R, Breakefield XO (2005) Codon-optimized Gaussia luciferase cDNA for mammalian gene expression in culture and in vivo. Mol Ther 11:435–443
Article PubMed CAS Google Scholar
Acknowledgements
This work was funded by the Korean Science and Engineering Foundation (KOSEF) grant funded by the Korea government (MOST; No. 2007-02242), National R&D Program for Cancer Control of Ministry of Health & Welfare (0820320), and Seoul R&BD program. H.J. Kim was supported by the BK21 Project for Medicine, Dentistry, and Pharmacy in Korea (2007).
Author information
Authors and Affiliations
- Department of Nuclear Medicine, College of Medicine, Seoul National University, 28 Yeongon-dong, Chongno-gu, Seoul, 110-744, South Korea
Hyun Joo Kim, June-Key Chung, Do Won Hwang, Dong Soo Lee & Soonhag Kim - Laboratory of Molecular Imaging and Therapy of Cancer Research Institute, College of Medicine, Seoul National University, Seoul, South Korea
Hyun Joo Kim, June-Key Chung & Do Won Hwang - Medical Research Center, College of Medicine, Seoul National University, Seoul, South Korea
Soonhag Kim
Authors
- Hyun Joo Kim
You can also search for this author inPubMed Google Scholar - June-Key Chung
You can also search for this author inPubMed Google Scholar - Do Won Hwang
You can also search for this author inPubMed Google Scholar - Dong Soo Lee
You can also search for this author inPubMed Google Scholar - Soonhag Kim
You can also search for this author inPubMed Google Scholar
Corresponding authors
Correspondence toJune-Key Chung or Soonhag Kim.
Rights and permissions
About this article
Cite this article
Kim, H.J., Chung, JK., Hwang, D.W. et al. In Vivo Imaging of miR-221 Biogenesis in Papillary Thyroid Carcinoma.Mol Imaging Biol 11, 71–78 (2009). https://doi.org/10.1007/s11307-008-0188-6
- Received: 26 May 2008
- Revised: 05 August 2008
- Accepted: 11 August 2008
- Published: 22 November 2008
- Issue Date: March 2009
- DOI: https://doi.org/10.1007/s11307-008-0188-6