Application of superparamagnetic iron oxide labeled antisense oligodeoxynucleotide probe in cellular magnetic resonance imaging.
- Author:
Ming WEN
1
;
Bi-Bo LI
;
Yu OUYANG
;
Ming-Dong JIANG
;
Yi LUO
;
Shao-Lin LI
Author Information
- Publication Type:Journal Article
- MeSH: Cell Line, Tumor; DNA, Antisense; chemistry; genetics; Ferric Compounds; chemistry; Humans; Magnetic Resonance Imaging; Magnetics; Molecular Probe Techniques; Oligodeoxyribonucleotides; chemistry; genetics; Oxyphil Cells; chemistry; Receptor, ErbB-2; analysis; genetics
- From: Acta Academiae Medicinae Sinicae 2008;30(5):554-558
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo prepare the superparamagnetic iron oxide (SPIO)-labeled antisense oligodeoxynucleotide (ASODN) probe and evaluate the application of this probe in cellular magnetic resonance imaging (MRI).
METHODSWe prepared the SPIO-labeled ASODN probe using chemical cross linking method to conjugate SPIO to ASODN, detected its configuration by atomic force microscopy, determined the conjugating rate and biology activation by high performance liquid chromatography, and detected the stability by polyacrylamide gel electrophoresis. After that, we transfected the SK-Br3 oncocytes which had over-expression of the c-erbB2 oncogene by this probes, observed the intracellular iron distribution by optical microscope, measured iron content by atomic absorption spectroscopy, and observed the signal change by MRI.
RESULTSAtomic force microscope showed that the SPIO-labeled ASODN probe was mostly spherical and well-distributed, with a diameter of 25-40 nm and a conjugating rate of 100%. This probe had inhered biological activity and stability. In addition, light microscopy revealed an intracellular uptake of iron oxides in the transfected SK-Br3 oncocyte, and the iron content of the group of transfected SK-Br3 oncocytes was significantly higher than those of other contrast groups (all P < 0.01). MRI showed that transfected SK-Br3 oncocyte had the lowest signal among all other cells (all P < 0.05).
CONCLUSIONSWe prepared the SPIO-labeled ASODN probe successfully. It can effectively transfect SK-Br3 oncocyte and enter SK-Br3 oncocyte, and thus reduce the signal intension in MRI.