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Preparation of liposome-coated oligonucleotide labeled with ^(99m)Tc and its uptake in vascular smooth muscle cells

Preparation of liposome-coated oligonucleotide labeled with ^(99m)Tc and its uptake in vascular smooth muscle cells
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摘要 To explore the preparation method of liposome-coated 99mTc-labeled antisense oligonucleotide (ASON), targeteing the proliferating cell nuclear antigen (PCNA), and to explore the biological characteristics and the uptake kinetics of a radiolabeled probe in vascular smooth muscle cells, an 18-base single-stranded antisense oligonucleotide targeting PCNA mRNA and the complementary strand (sense oligonucleotide, SON) were synthesized. The ASON (SON) was labeled with 99mTc, by conjugating the bifunctional chelator (hydrazino nicotinamide, HYNIC), and puri- fied through a gel filtration column of Sephadex G-25. The product was then encapsulated in cationic liposome (oli- gofectamineTM). The radiolabeling efficiency, radiochemical purity, stability of the liposome-coated 99mTc-HYNIC- ASON in a phosphate buffered solution (PBS), and fresh human serum and its uptake rate were studied. There was no significant difference between the 99mTc radiolabeling efficiencies of HYNIC-ASON and HYNIC-SON, which were 60.04% ± 1.92% and 59.60% ± 2.53%, respectively (P > 0.05, n = 5). The radiochemical purity of the lipo- some-coated 99mTc-HYNIC-ASON was 94.70% ± 1.90% (n = 5). And after incubation with PBS and fresh human se- rum at a concentration of 1.2 μg·mL-1 for 120 min, the radiochemical purities were 92.27% and 91.55% respectively. At 90 min after transfection, the uptake rate of the liposome-coated 99mTc-HYNIC-ASON reached its peak of 83.8% ± 5.92% in vascular smooth muscle cells (VSMCs) and was much higher than that of the nonliposome-coated 99mTc- HYNIC-ASON, which was 11.16% ± 0.54% (P < 0.01, n = 4). The labeling method of PCNA ASON (SON) conju- gated by HYNIC has been proved successful. The liposome was able to enhance the ASON (SON) uptake in VSMCs, and could be widely used as a safe, convenient, effective gene transfer carrier. To explore the preparation method of liposome-coated ^99mTc-labeled antisense oligonucleotide (ASON), targeteing the proliferating cell nuclear antigen (PCNA), and to explore the biological characteristics and the uptake kinetics of a radiolabeled probe in vascular smooth muscle cells, an 18-base single-stranded antisense oligonucleotide targeting PCNA mRNA and the complementary strand (sense oligonucleotide, SON) were synthesized. The ASON (SON) was labeled with ^99mTc, by conjugating the bifunctional chelator (hydrazino nicotinamide, HYNIC), and purified through a gel filtration column of Sephadex G-25. The product was then encapsulated in cationic liposome (oligofectamineTM). The radiolabeling efficiency, radiochemical purity, stability of the liposome-coated ^99Tc-HYNICASON in a phosphate buffered solution (PBS), and fresh human serum and its uptake rate were studied. There was no significant difference between the ^99mTc radiolabeling efficiencies of HYNIC-ASON and HYNIC-SON, which were 60.04% ± 1.92% and 59.60% ± 2.53%, respectively (P 〉 0.05, n = 5). The radiochemical purity of the liposome-coated &99mTc-HYNIC-ASON was 94.70% ± 1.90% (n = 5). And after incubation with PBS and fresh human serum at a concentration of 1.2 μg·mL^-1 for 120 min, the radiochemical purities were 92.27% and 91.55% respectively. At 90 min after transfection, the uptake rate of the liposome-coated ^99mTc-HYNIC-ASON reached its peak of 83.8% ± 5.92% in vascular smooth muscle cells (VSMCs) and was much higher than that of the nonliposome-coated ^99mTc- HYNIC-ASON, which was 11.16% ± 0.54% (P 〈 0.01, n = 4). The labeling method of PCNA ASON (SON) conjugated by HYNIC has been proved successful. The liposome was able to enhance the ASON (SON) uptake in VSMCs, and could be widely used as a safe, convenient, effective gene transfer carder.
出处 《Nuclear Science and Techniques》 SCIE CAS CSCD 2007年第2期101-106,共6页 核技术(英文)
基金 Supported by Natural Science Foundation of China (No.30271439)
关键词 放射性同位素标记 脂质体包覆寡核苷酸 锝99M 制备 血管平滑肌细胞 吸收 PCNA Proliferating cell nuclear antigen, Oligonucleotides, Antisense, Isotope labeling, Liposome, Vascularsmooth muscle cells
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参考文献12

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