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J Nucl Med. 2008; 49 (Supplement 1):168P
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Novel Approaches to Molecular Imaging

Novel Approaches to Molecular Imaging Posters

In vivo imaging of the biogenesis and functional targeting of miR-221 in papillary thyroid carcinoma

Hyun Joo Kim1, Soonhag Kim1, Young Ha Kim1, June-Key Chung1, Dong Soo Lee1 and Keon Wook Kang1

1 Nuclear Medicine, Seoul National University College of Medicine, Seoul, South Korea


Formula

729

Objectives: MicroRNAs (miRNAs) are small noncoding RNA molecules that control expression of target genes. The abnormally expressed miRNAs function as oncogenes or tumor suppressors in human cancer. To evaluate the quantities and gene regulation of miR-221 in papillary thyroid carcinoma (PTC), we performed qRT-PCR and microarray and developed Gaussia luciferase (Gluc) reporter system regulated by miR-221.

Methods: The quantities of primary or mature miR-221 in cells were measured with qRT-PCR. CMV/Gluc-3xPT including 3 repeat perfect complementary target sequences (3xPT) of miR-221 was transfected into cells and pre.miR-221 or anti.miR-221 were co-transfected with CMV/Gluc-3xPT. The Gluc activities in cells were measured by luciferase assay. Mice implanted with PTC expressing Gluc regulated by miR-221 were monitored with bioluminescence imaging for 6 days. Microarray was performed with 44K probes and total RNA isolated from pre.miR-221 or anti.miR-221 treated thyroid cells. Target genes regulated by miR-221 were evaluated with RT-PCR and Gluc reporter system.

Results: PCR results showed primary or mature miR-221 is 2.24 or 17 times higher in PTC than in normal thyroid cells. The Gluc activities in cells were repressed more than 2 times by endogenous miR-221 or exogenous pre.miR-221 or recovered by anti.miR-221 in vitro and in vivo. Microarray analysis showed thousands of genes were directly and indirectly regulated by miR-221 and shifted gene expression pattern of normal thyroid cells toward PTC. Of the miR-221 directly targeted genes, HOXB5, which contains miR-221 seed region in its 3'UTR, was significantly down-regulated by miR-221.

Conclusions: MiR-221 is highly expressed in PTC and it drives carcinoma gene expression patterns by directly and indirectly regulating numerous genes, including HOXB5.





This Article
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Right arrow Email this article to a friend
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Right arrow Alert me to new issues of the journal
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Right arrow Articles by Kim, H. J.
Right arrow Articles by Kang, K. W.
PubMed
Right arrow Articles by Kim, H. J.
Right arrow Articles by Kang, K. W.