[1]李佳,王政杰,张磊,等.177Lu-DOTA-anti-CITED1-PNA的制备、鉴定及其对甲状腺乳头状癌K1细胞增殖能力的影响[J].第三军医大学学报,2019,41(20):1954-1960.
 LI Jia,WANG Zhengjie,ZHANG Lei,et al.Preparation and identification of 177Lu-labeled DOTA-anti-CITED1-PNA and its effect on proliferation of human papillary thyroid cancer K1 cells[J].J Third Mil Med Univ,2019,41(20):1954-1960.
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177Lu-DOTA-anti-CITED1-PNA的制备、鉴定及其对甲状腺乳头状癌K1细胞增殖能力的影响(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
41卷
期数:
2019年第20期
页码:
1954-1960
栏目:
基础医学
出版日期:
2019-10-30

文章信息/Info

Title:
Preparation and identification of 177Lu-labeled DOTA-anti-CITED1-PNA and its effect on proliferation of human papillary thyroid cancer K1 cells
作者:
李佳王政杰张磊王英黄欢曹熠熠庞华
重庆医科大学附属第一医院核医学科
Author(s):
LI Jia WANG Zhengjie ZHANG Lei WANG Ying HUANG Huan CAO Yiyi PANG Hua

Department of Nuclear Medicine, First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China

关键词:
甲状腺乳头状癌CITED1放射性核素镥反义肽核酸
Keywords:
papillary thyroid carcinoma CITED1 Lutetium-177 antisense peptide nucleic acid
分类号:
R445.5;R736.1;R817.5
文献标志码:
A
摘要:

目的 研究放射性镥核素(177Lu)标记的结合转化激活因子(Cbp/p300 interacting transactivator with Glu/Asp rich carboxy-terminal domain 1,CITED1)mRNA反义肽核酸(antisense peptide nucleic acid,asPNA)即177Lu-DOTA-anti-CITED1-PNA的制备,鉴定标记物的理化性质并探讨其在人甲状腺乳头状癌K1细胞系中的摄取能力及细胞毒性作用。方法 实时荧光定量PCR(RT-PCR)鉴定肽核酸(peptide nucleic acid,PNA)的靶向特异性。通过间接标记法得到反义探针177Lu-DOTA-anti-CITED1-PNA并测定其标记率,检测其体内外的稳定性。体外细胞实验测定反义探针的摄取能力及对K1细胞增殖能力影响。结果 RT-PCR显示反义肽核酸作用于K1细胞后能明显降低CITED1的表达;反义探针的放射化学纯度为(94.5±0.12)%,比活度为(8.7±0.53)MBq/μg,48 h的放射化学纯度仍大于90%;体外细胞摄取、滞留以及细胞增殖实验显示反义探针能被人K1细胞特异性摄取,且相较于未偶联的177Lu和asPNA,177Lu和asPNA偶联后联用具有显著的抗肿瘤细胞作用。结论 成功制备了177Lu标记的CITED1mRNA反义肽核酸探针,证实反义探针对K1细胞具有特异性、靶向性且反义探针的联合作用能有效降低细胞活力及增殖能力。

Abstract:

Objective To prepare and characterize 177Lu-DOTA-anti-CIITED1-PNA, a 177Lu-labled antisense probe for the antisense peptide nucleic acid (asPNA) of Cbp/p300 interacting transactivator with Glu/Asp rich carboxy-terminal domain 1 (CITED1) mRNA and assess its uptake and cytotoxicity in human papillary thyroid cancer K1 cells.  MethodsThe targeting specificity of the peptide nucleic acid was identified by RT-PCR. The antisense probe 177Lu-DOTA-anti-CITED1-PNA was obtained using an indirect labeling method, and the labeling rate and in vitro and in vivo stability of the probe were assessed. The uptake of the antisense probe and its effect on the proliferation of K1 cells in vitro were evaluated.  ResultsRT-PCR showed that the asPNA could significantly reduce the expression of CITED1 in K1 cells. The prepared antisense probe had a radiochemical purity of (94.5±0.12)% with a specific activity of 3.7±0.53 MBq/μg; its radiochemical purity was still above 90% after 48 h. This antisense probe could be specifically taken up by human K1 cells, and compared with 177Lu-labeled DOTA and a nonsense-PNA, the antisense probe 177Lu-DOTA-anti-CITED1-PNA showed a much stronger anti-tumor effect in K1 cells.  ConclusionWe successfully obtained 177Lu-labeled CITED1 asPNA probe, which can specifically target K1 cells and effectively reduce the cell viability and proliferation in vitro.

参考文献/References:

[1]SCHMIDT A, IGLESIAS L, KLAIN M, et al. Radioactive iodine-refractory differentiated thyroid cancer: an uncommon but challenging situation[J]. Arch Endocrinol Metab, 2017, 61(1): 81-89. DOI: 10.1590/2359-3997000000245.
[2]DASH A, PILLAI M R, KNAPP F F Jr. Production of 177Lu for targeted radionuclide therapy: available options[J]. Nucl Med Mol Imaging, 2015, 49(2): 85-107. DOI: 10.1007/s13139-014-0315-z.
[3]SAKAI J, MAEDA T, TARUMOTO N, et al. Corrigendum to “A novel detection procedure for mutations in the 23S rRNA gene of Mycoplasma pneumoniae with peptide nucleic acid-mediated loop-mediated isothermal amplification assay”[J]. J Microbiol Methods, 2018, 144: 192. DOI: 10.1016/j.mimet.2017.11.004.
[4]LI H, GUAN H, GUO Y, et al. CITED1 promotes proliferation of papillary thyroid cancer cells via the regulation of p21 and p27[J]. Cell Biosci, 2018, 8(1): 57-63. DOI: 10.1186/s13578-018-0256-9.
[5]CAMACHO X, CALZADA V, FERNANDEZ M, et al. 177Lu-DOTA-bevacizumab: radioimmunotherapy agent for melanoma[J]. Curr Radiopharm, 2017, 10(1): 21-28. DOI: 10.2174/1874471009666161010155246.
[6]BREEMAN W A, CHAN H S, DE ZANGER R M, et al. Overview of development and formulation of 177Lu-DOTA-TATE for PRRT[J]. Curr Radiopharm, 2016, 9(1): 8-18.
[7]LI W B, ZHOU J, XU L, et al. Identification of genes associated with papillary thyroid carcinoma (PTC) for diagnosis by integrated analysis[J]. Horm Metab Res, 2016, 48(4): 226-231. DOI: 10.1055/s-0035-1569289.
[8]KAOCHAR S, DONG J, TORRES M, et al. ICG-001 exerts potent anticancer activity against uveal melanoma cells[J]. Invest Ophthalmol Vis Sci, 2018, 59(1): 132-143. DOI: 10.1167/iovs.17-22454.
[9]PHELPS H M, AL-JADIRY M F, CORBITT N M, et al. Molecular and epidemiologic characterization of Wilms tumor from Baghdad, Iraq[J]. World J Pediatr, 2018, 14(6): 585-593. DOI: 10.1007/s12519-018-0181-3.
[10]WOLF K I, JHA A, VAN BERKEL A, et al. Eruption of metastatic paraganglioma after successful therapy with 177Lu/90Y-DOTATOC and 177Lu-DOTATATE[J]. Nucl Med Mol Imaging, 2019, 53(3): 223-230. DOI: 10.1007/s13139-019-00579-w.
[11]BALLINGER J R. Theranostic radiopharmaceuticals: established agents in current use[J]. Br J Radiol, 2018, 91(1091): 20170969. DOI: 10.1259/bjr.20170969.
[12]LIU D, BALKIN E R, JIA F, et al. Targeted antisense radiotherapy and dose fractionation using a 177Lu-labeled anti-bcl-2 peptide nucleic acid-peptide conjugate[J]. Nucl Med Biol, 2015, 42(9): 704-710. DOI: 10.1016/j.nucmedbio.2015.05.006.
[13]PANSUWAN H, DITMANGKLO B, VILAIVAN C, et al. Hydrophilic and cell-penetrable pyrrolidinyl peptide nucleic acid via post-synthetic modification with hydrophilic side chains[J]. Bioconjug Chem, 2017, 28(9): 2284-2292. DOI: 10.1021/acs.bioconjchem.7b00308.
[14]刘春冬, 王建华, 曾芳.修饰性肽核酸的细胞转运[J].生物工程学报, 2016, 32(3): 292-305. DOI: 10.13345/j.cjb.150275.
LIU C D, WANG J H, ZENG F. Cellular delivery of modified peptide nucleic acids: a review[J]. Chin J Biotechnol, 2016, 32(3): 292-305. DOI: 10.13345/j.cjb.150275.
[15]HOFVING T, SANDBLOM V, ARVIDSSON Y, et al. 177Lu-octreotate therapy for neuroendocrine tumours is enhanced by Hsp90 inhibition[J]. Endocr Relat Cancer, 2019, 26(4): 437-449. DOI: 10.1530/ERC-18-0509.
 

更新日期/Last Update: 2019-10-25