In vivo imaging of miR-221 biogenesis in papillary thyroid carcinoma.
OBJECTIVE
To investigate the overexpression of miR-221 in papillary thyroid carcinoma (PTC), we developed a Gaussia luciferase (Gluc) system regulated by miR-221.


METHODS
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.
Kim
Hyun Joo
HJ
Department of Nuclear Medicine, College of Medicine, Seoul National University, 28 Yeongon-dong, Chongno-gu, Seoul, 110-744, South Korea.
Chung
June-Key
JK
Hwang
Do Won
DW
Lee
Dong Soo
DS
Kim
Soonhag
S
D000818:Animals; D002291:Carcinoma, Papillary / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D045744:Cell Line, Tumor; D003587:Cytomegalovirus / Q000235:genetics / Q000378:metabolism; D003952:Diagnostic Imaging / Q000379:methods*; D020869:Gene Expression Profiling; D017930:Genes, Reporter; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008819:Mice, Nude; D035683:MicroRNAs / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D009368:Neoplasm Transplantation; D016133:Polymerase Chain Reaction; D013961:Thyroid Gland / Q000378:metabolism; D013964:Thyroid Neoplasms / Q000235:genetics / Q000378:metabolism*; D051598:Whole Body Imaging / Q000379:methods
D008156:Luciferases; D035683:MicroRNAs; C525709:MIR221, human