~(99m)Tc-GE11分子探针在荷人肺腺癌裸鼠体内分布及受体显像研究
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摘要
目的制备靶向肺腺癌细胞跨膜表达蛋白表皮生长因子受体(epidermal growth factor receptor,EGFR)的核素分子探针~(99m)Tc-GE11,鉴定其理化性质并探讨其在荷瘤裸鼠体内分布及显像情况。方法采用氯化亚锡氧化还原法对GE11进行~(99m)Tc标记,初步鉴定其标记率、比活度、放化纯度及稳定性等,并观察多肽标记物在荷瘤裸鼠体内的生物分布及显像情况。结果 ~(99m)Tc-GE11标记率为(93.12±0.83)%,比活度为(24.21±0.42)TBq/mmol。体外室温放置6、12 h后,其放化纯度仅下降了2.14%、3.11%。Sephadex G50柱层析后,其与血浆蛋白结合率为4.8%;与不同浓度半胱氨酸37℃温育1 h后,未结合~(99m)Tc含量无明显变化;脂/水分配系数lgP为(-1.87±0.05);荷瘤裸鼠体内生物分布及SPECT显像示:~(99m)Tc-GE11在血液、肾脏中的放射性清除迅速,肿瘤组织可以选择性浓聚~(99m)Tc-GE11,在注射1.0 h时,肿瘤/肌肉比值达到最大,且肿瘤组织显影最清晰。结论制备的~(99m)Tc-GE11小分子多肽探针理化特性理想,在荷人肺腺癌裸鼠模型中显像较好。
Objective To prepare a ~(99 m)Tc-labled molecular probe(~(99 m)Tc-GE11) targeting epidermal growth factor receptor(EGFR) on lung adenocarcinoma cells and evaluate its biodistribution and performance for EGFR imaging in nude mice bearing lung adenocarcinoma xenografts. Methods GE11 was labeled with ~(99 m)Tc through oxidation and reduction of stannous chloride, and the labeling rate, specific activity, radiochemical purity and stability of ~(99 m)Tc-GE11 were assessed. The biodistribution profile and receptor imaging performance of ~(99 m)Tc-GE11 were evaluated in a nude mouse model bearing lung adenocarcinoma xenografts. Results The mean labeling rate of ~(99 m)TC-GE11 was(93.12±1.43)%, and its mean specific activity was 24.21±0.42 TBq/mmol. At room temperature, the radiochemical purity of ~(99 m)TC-GE11 was reduced by 2.14% at 6 h and by 3.11% at 12 h. The results of chromatography with Sephadex-G50 column showed that the binding rate of ~(99 m)TC-GE11 with plasma proteins was 4.8%. At 37 ℃, incubation of ~(99 m)Tc-GE11 with different concentrations of cysteine for 1 h did not cause significant changes in the level of free ~(99 m)Tc. The fat/water distribution coefficient(lgP) of ~(99 m)TC-GE11 was-1.87±0.05. In the tumor-bearing nude mice, ~(99 m)Tc-GE11 was rapidly cleared in blood and kidney after intravenous injection and was selectively accumulated in the tumor; the tumor/non-tumor(T/NT) ratio reached the highest level and the clearest tumor imaging was achieved at 1 h after the injection. Conclusion ~(99 m)Tc-GE11 molecular probe prepared based on a small molecular polypeptide possesses good physiochemical properties with an optimal biodistribution profile, and allows clear in vivo tumor imaging in tumor-bearing nude mice.
Objective To prepare a ~(99 m)Tc-labled molecular probe(~(99 m)Tc-GE11) targeting epidermal growth factor receptor(EGFR) on lung adenocarcinoma cells and evaluate its biodistribution and performance for EGFR imaging in nude mice bearing lung adenocarcinoma xenografts. Methods GE11 was labeled with ~(99 m)Tc through oxidation and reduction of stannous chloride, and the labeling rate, specific activity, radiochemical purity and stability of ~(99 m)Tc-GE11 were assessed. The biodistribution profile and receptor imaging performance of ~(99 m)Tc-GE11 were evaluated in a nude mouse model bearing lung adenocarcinoma xenografts. Results The mean labeling rate of ~(99 m)TC-GE11 was(93.12±1.43)%, and its mean specific activity was 24.21±0.42 TBq/mmol. At room temperature, the radiochemical purity of ~(99 m)TC-GE11 was reduced by 2.14% at 6 h and by 3.11% at 12 h. The results of chromatography with Sephadex-G50 column showed that the binding rate of ~(99 m)TC-GE11 with plasma proteins was 4.8%. At 37 ℃, incubation of ~(99 m)Tc-GE11 with different concentrations of cysteine for 1 h did not cause significant changes in the level of free ~(99 m)Tc. The fat/water distribution coefficient(lgP) of ~(99 m)TC-GE11 was-1.87±0.05. In the tumor-bearing nude mice, ~(99 m)Tc-GE11 was rapidly cleared in blood and kidney after intravenous injection and was selectively accumulated in the tumor; the tumor/non-tumor(T/NT) ratio reached the highest level and the clearest tumor imaging was achieved at 1 h after the injection. Conclusion ~(99 m)Tc-GE11 molecular probe prepared based on a small molecular polypeptide possesses good physiochemical properties with an optimal biodistribution profile, and allows clear in vivo tumor imaging in tumor-bearing nude mice.
引文
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[2] PASSIGLIA F,RIZZO S,DI MAIO M,et al.The diagnostic accuracy of circulating tumor DNA for the detection of EGFR-T790M mutation in NSCLC:A systematic review and meta-analysis[J].Sci Rep,2018,8(1):13379.DOI:10.1038/s41598-018-30780-4.
[3] XU L,HAUSMANN M,DIETMAIER W,et al.Expression of growth factor receptors and targeting of EGFR incholangiocarcinoma cell lines[J].BMC Cancer,2010,10:302.DOI:10.1186/1471-2407-10-302.
[4] WANG J S,WANG G B,LI B,et al.MiR-141-3p is a key negative regulator of the EGFR pathway in osteosarcoma[J].Oncotargets Ther,2018,11:4461-4478.DOI:10.2147/OTT.S171304.
[5] SHEN J,ZHANG T,CHENG Z,et al.Lycorine inhibits glioblastoma multiforme growth through EGFR suppression[J].J Exp Clin Cancer Res,2018,37(1):157.DOI:10.1186/s13046-018-0785-4.
[6] MENG Q Q,LI Z.Molecular imaging probes for diagnosis and therapy evaluation of breast cancer[J].Int J Biomed Imaging,2013,2013:230487.DOI:10.1155/2013/230487.
[7] LI Z H,ZHAO R J,WU X H,et al.Identification and characterization of a novel peptide ligand of epidermal growth factor receptor for targeted delivery oftherapeutics[J].FASEB J,2005,19(14):1978-1985.DOI:10.1096/fj.05-4058com.
[8] 郭妍,李宗海,顾健人,等.新多肽配体GE11与表皮生长因子受体的结合能力分析[J].分析测试学报,2008,27(12):1318-1321.DOI:10.3969/j.issn.1004-4957.2008.12.012.GUO Y,LI Z H,GU J R.et al.Study on the binding ability of new peptide ligand GE11 and EGFR[J].J Instrum Anal,2008,27(12):1318-1321.DOI:10.3969/j.issn.1004- 4957.2008.12.012.
[9] 查林,冯世斌,郑磊,等.整合素αvβ3放射性配体99Tcm-TP1326的制备及其正常兔显像研究[J].第三军医大学学报,2012,34(3):235-238.DOI:10.16016/j.1000-5404.2012.03.024.CHA L,FENG S B,ZHENG L,et al.Preparation of 99Tcm-TP1326 targeting integrin αvβ3 receptor and its imaging in rabbits[J].J Third Mil Med Univ,2012,34(3):235-238.DOI:10.16016/j.1000-5404.2012.03.024.
[10] 丁小江,郑磊,黄定德,等.99Tcm标记VPAC1 配体TP1724 及其在动物体内分布与显像研究[J].第三军医大学学报,2016,38(19):2107-2113.DOI:10.16016 /j.1000-5404.201603049 DING X J,ZHENG L,HUANG D D,et al.Distribution and imaging of a 99Tcm-labeled linear peptide derived from vasoactive intestinal peptide receptor in vivo[J].J Third Mil Med Univ,2016,38(19):2107-2113.DOI:10.16016 /j.1000-5404.201603049.
[11] 邹丹丹,窦骏.恶性肿瘤的分子靶点检测和靶向治疗[J].临床与实验病理学杂志,2012,28(9):1026-1029.DOI:10.13315/j.cnki.cjcep.2012.09.035 ZOU D D,DOU J.Molecular target detection and targeted therapy for malignant tumors [J].Chin J Clin Exp Pathol,2012,28(9):1026-1029.DOI:10.13315/j.cnki.cjcep.2012.09.035.
[12] XU N Q,FANG W F,MU L B,et al.Overexpression of wildtype EGFR is tumorigenic and denotes a therapeutic target in non-small cell lung cancer[J].Oncotarget,2016,7(4):3884-3896.DOI:10.18632/oncotarget.6461.
[13] LEE M.Is EGFR expression important in non-small cell lung cancer?[J].Thorax,2006,61(2):98-99.DOI:10.1136/thx.2005.047936.
[14] CONSTANTINOU C,PAPADOPOULOS S,KARYDA E,et al.Expression and clinical significance of claudin-7,PDL-1,PTEN,c-kit,c-met,c-myc,ALK,CK5/6,CK17,p53,EGFR,ki67,p63 in triple-negative breast cancer-A single centre prospective observational study[J].In Vivo,2018,32(2):303-311.DOI:10.21873/invivo.11238.
[15] LOUD J T,PETERS J A,FRASER M,et al.Applications of advances in molecular biology and genomics to clinical cancer care[J].Cancer Nurs,2002,25(2):110-122.
[16] HOPKINSA L,GROOM C R.The druggable genome[J].Nat Rev Drug Discov,2002,1(9):727-730.DOI:10.1038/nrd892.
[17] GOLDENBERG D M.Advancing role of radiolabeled antibodies in the therapy of cancer[J].Cancer Immunol,Immunother,2003,52(5):281-296.
[18] TARHINI A A,KIRKWOOD J M.Clinical and immunologic basis of interferon therapy in melanoma[J].Ann N Y Acad Sci,2009,1182:47-57.DOI:10.1111/j.1749-6632.2009.05073.x.
[19] 李宗海.表皮生长因子受体介导的靶向性基因治疗导入系统[D].上海:复旦大学,2005.LI Z H.Epidermal growth factor receptor-targeted gene delivery system[D].Shanghai:Fudan University,2005.
[20] 李贵平,黄宝丹,杜丽,等.99mTc标记的新型肺癌多肽分子探针及其生物学分布[J].南方医科大学学报,2013,33(8):1169-1172.DOI:10.3969/j.issn.1673-4254.2013.08.15.LI G P,HUANG B D,DU L,et al.99mTc radiolabeling of a novel polypeptide molecular probe for lung cancer and its biodistribution in animals[J].J South Med Univ,2013,33(8):1169-1172.DOI:10.3969/j.issn.1673-4254.2013.08.15.