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  • Title: Noninvasive imaging of human telomerase reverse transcriptase (hTERT) messenger RNA with 99mTc-radiolabeled antisense probes in malignant tumors.
    Author: Liu M, Wang RF, Zhang CL, Yan P, Yu MM, Di LJ, Liu HJ, Guo FQ.
    Journal: J Nucl Med; 2007 Dec; 48(12):2028-36. PubMed ID: 18006621.
    Abstract:
    UNLABELLED: The expression of human telomerase reverse transcriptase (hTERT) is present in most malignant cells (>85%) but is undetectable in most normal somatic cells. Visualization of hTERT expression using radionuclide targeting can provide important diagnostic information in malignant tumors. The overall aim of this study was to evaluate whether (99m)Tc-radiolabeled antisense oligonucleotide (ASON) targeting hTERT messenger RNA (mRNA) can be used for imaging of hTERT expression in vivo. METHODS: One 18-mer antisense or sense uniformly phosphorothioate-modified oligonucleotide targeting hTERT mRNA was radiolabeled with (99m)Tc through the bifunctional chelator N-hydroxysuccinimidyl derivative of S-acetylmercaptoacetyltriglycine (S-acetyl NHS-MAG3). Then the radiolabeled probe was characterized in vitro. Reverse transcription-polymerase chain reaction (RT-PCR) was performed to assay the mRNA level after proliferating cells had been incubated with the antisense and sense probes. (99m)Tc-MAG3-ASON or (99m)Tc-MAG3-SON was injected intravenously in mammary tumor-bearing BALB/c nude mice. Biodistribution and in vivo imaging was performed periodically. All data were analyzed by statistical software. RESULTS: The labeling efficiencies of (99m)Tc-MAG3-ASON/SON reached 76% +/- 5% (n = 5) within 15-30 min at room temperature, the specific activity was up to 1,850 kBq/mug, and the radiochemical purity was >96% after purification. (99m)Tc-MAG3-ASON showed complete stability at room temperature and in fresh 37 degrees C human serum. In comparison with (99m)Tc-MAG3-SON, (99m)Tc-MAG3-ASON preserved the capacity to bind living hTERT-expressing cells specifically and to inhibit the expression of hTERT mRNA significantly as well as ASON. In nude mice bearing hTERT-expressing MCF-7 xenografts, tumor radioactivity uptake of (99m)Tc-MAG3-ASON after injection was significantly higher than that of (99m)Tc-MAG3-SON after injection (P < 0.05). The hTERT-expressing xenografts were clearly imaged at 4-8 h noninvasively after injection of (99m)Tc-MAG3-ASON, whereas the xenografts were not imaged at any time after injection of (99m)Tc-MAG3-SON. CONCLUSION: This in vivo study provides evidence that (99m)Tc-MAG3-ASON targeting hTERT mRNA can be used as a potential candidate for visualization of hTERT expression in carcinomas.
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