99Tcm标记小干扰RNA探针的干扰活性及其在荷瘤鼠体内的生物分布
In-vitro Inhibitory Activity and In-vivo Biodistribution of 99Tcm Radiolabeled Small Interference RNA
-
摘要: 使用双功能螯合剂NHSMAG3和氯化亚锡还原法构建99Tcm标记的小干扰RNA（Small Interference RNA，siRNA）探针。将标记和未标记的端粒逆转录酶（Human Telomerase Reverse Trancriptase，hTERT）靶向siRNA转染至肝癌HepG2细胞，72 h后，蛋白印迹法显示两者具有相近的蛋白抑制率（约76.7%）。标记物在荷HepG肿瘤裸鼠体内的生物分布显示，肾脏分布最高，其次为肝脏。注射hTERT靶向探针后 1～6 h内，肿瘤分布由（0.82 ± 0.16）%ID/g增加至（0.97 ± 0.15）%ID/g，肿瘤与血液和肿瘤与肌肉的放射性摄取比（T/NT）分别为2.62 ± 0.70和6.02 ± 0.52，显著高于对照组（P< 0.05）。以上结果提示，99Tcm-siRNA探针对活体肿瘤示踪具有良好的研究前景和潜在的应用价值。Abstract: Small interference RNA (siRNA) was radiolabeled by using the bifunctional chelator of NHS-MAG3 and SnCl2●2H2O. After transfected into Hepatocarcinoma HepG2 cells, labeled and unlabeled hTERTtargeted siRNA had the similar inhibitory rate about 76.7% measured by western blotting method. In the biodistribution study, the radio-active accumulation was primarily found in the kidneys, and then in liver. The radioactivity of hTERT-targeted siRNA in tumor increased from (0.82 ± 0.16) %ID/g to (0.97 ± 0.15) %ID/g from 1 to 6 hours after the administration. The uptake ratio of tumor to blood and tumor to muscle were 2.62 ± 0.70 and 6.02 ± 0.52, respectively, significantly higher than that of the control group (P<0.05). These results indicated that 99Tcm radiolabeled hTERTtargeted siRNA allows for prospective future and potential application value in the noninvasive visualization of tumor telomerase in vivo.
-
Keywords:
- 99Tcm /
- small interference RNA (siRNA) /
- inhibitory activity /
- biodistribution
-
-
[1] 康磊, 王荣福. 人端粒酶逆转录酶在肿瘤靶向分子显像中的研究进展 [J]. 中华核医学杂志, 2010, 34(1): 68-71. [2] Liu M, Wang RF, Zhang CL, et al. Noninvasive imaging of human telomerase reverse transcriptase (hTERT) messenger RNA with 99Tcm radiolabeled antisense probes in malignant tumors [J]. J Nucl Med, 2007, 48(12): 2 028-2 036. [3] 康磊, 王荣福, 闫平, 等. 99Tcm标记的小干扰RNA探针的制备及稳定性评价[J]. 核化学与放射化学, 2010, 32(6): 348-353. [4] Masutomi K, Yu EY, Khurts S, et al. Telomerase maintains telomere structure in normal human cells [J]. Cell, 2003, 114(2): 241-253. [5] Bramsen JB, Laursen MB, Nielsen AF, et al. A large-scale chemical modification screen identifies design rules to generate siRNAs with high activity, high stability and low toxicity [J]. Nucleic Acids Res, 2009, 37(9): 2 867-2 881. [6] 刘萌, 王荣福. S-乙酰基-NHS-MAG3在反义显像研究中的应用及进展[J]. 同位素, 2004, 17(3): 164-168. [7] van de Water FM, Boerman OC, Wouterse AC, et al. Intravenously administered short interfering RNA accumulates in the kidney and selectively suppresses gene function in renal proximal tubules [J]. Drug Metab Dispos, 2006, 34(8): 1 393-1 397. [8] McCaffrey AP, Meuse L, Pham TT, et al. RNA interference in adult mice [J]. Nature, 2002, 418(6 893): 38-39.
计量
- 文章访问数: 452
- HTML全文浏览量: 0
- PDF下载量: 1065
下载:
