近红外荧光染料IR-780在肾透明细胞癌中的成像特征
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摘要
背景:课题组研发了一组可以直接与肿瘤细胞结合的有机近红外荧光染料,IR-780是其中的优秀代表。目的:评估近红外荧光染料IR-780在肾透明细胞癌中的特异性成像效果。方法:取对数生长期的人肾透明细胞癌细胞786-O、ACHN和正常人胚肾细胞293T,分别与20μmol/L荧光染料IR-780共孵育30 min,激光共聚焦显微镜下观察染料在肾透明细胞癌细胞中的成像。取对数生长期的人肾透明细胞癌细胞786-O,与20μmol/L近红外荧光染IR-780共孵育30 min,激光共聚焦显微镜下观察染料在细胞线粒体与及溶酶体内的定位。在人血液样本中分别加入10,100,1 000,10 000个DAPI染色的人肾透明细胞癌细胞786-O,分离培养各组单个核细胞,再与20μmol/L近红外荧光染料IR-780共孵育30 min,激光共聚焦显微镜下观察DAPI和IR780双阳性染色细胞。结果与结论:(1)近红外荧光染料IR-780具备使多种肾透明细胞癌细胞显像的能力,对正常肾胚上皮细胞则无此能力;(2)近红外荧光染料IR-780与溶酶体或线粒体均有明显的染色重叠,验证了IR-780在膜性细胞器线粒体和溶酶体内的选择性聚集作用;(3)近红外荧光染料IR-780可检测到血液中微量的肾透明细胞癌细胞;(4)结果表明,近红外荧光染料IR-780能够在肾透明细胞癌细胞内特异性蓄积,可用于血液中肾透明细胞癌细胞的特异性诊断。
BACKGROUND: The near-infrared fluorescent dyes that directly bind to tumor cells have been developed, and IR-780 stands out. OBJECTIVE: To evaluate the specific imaging effect of near-infrared fluorescent dye IR-780 in clear cell renal cell carcinoma(ccRCC). METHODS: Human ccRCC cells 786-O, ACHN and normal human embryonic kidney 293 T cells in logarithmic growth phase were incubated with 20 μmol/L IR-780 for 30 minutes. The localization of the dye in ccRCC was observed by laser confocal microscopy. Human ccRCC cells 786-O in logarithmic phase was incubated with 20 μmol/L IR-780 for 30 minutes. The localization of the dye in mitochondria and lysosomes was observed under confocal laser scanning microscope. 10, 100, 1 000 and 10 000 DAPI-stained human ccRCC cells 786-O were respectively added to human blood samples. The mononuclear cells in each group were isolated and cultured with 20 μmol/L IR-780 for 30 minutes. DAPI and IR-780 double positive cells were observed under confocal laser scanning microscope. RESULTS AND CONCLUSION: IR-780 possessed the ability to visualize a variety of ccRCC cells, but not for normal renal epithelial cells. IR-780 had obvious overlapping staining in lysosomes or mitochondria, indicating its selective accumulation in mitochondria and lysosomes in the organelles. IR-780 could detect a small amount of ccRCC in the blood. These results suggest that the near-infrared fluorescent dye IR-780 can specifically accumulate in ccRCC cells, which can be used for specific diagnosis of ccRCC.
BACKGROUND: The near-infrared fluorescent dyes that directly bind to tumor cells have been developed, and IR-780 stands out. OBJECTIVE: To evaluate the specific imaging effect of near-infrared fluorescent dye IR-780 in clear cell renal cell carcinoma(ccRCC). METHODS: Human ccRCC cells 786-O, ACHN and normal human embryonic kidney 293 T cells in logarithmic growth phase were incubated with 20 μmol/L IR-780 for 30 minutes. The localization of the dye in ccRCC was observed by laser confocal microscopy. Human ccRCC cells 786-O in logarithmic phase was incubated with 20 μmol/L IR-780 for 30 minutes. The localization of the dye in mitochondria and lysosomes was observed under confocal laser scanning microscope. 10, 100, 1 000 and 10 000 DAPI-stained human ccRCC cells 786-O were respectively added to human blood samples. The mononuclear cells in each group were isolated and cultured with 20 μmol/L IR-780 for 30 minutes. DAPI and IR-780 double positive cells were observed under confocal laser scanning microscope. RESULTS AND CONCLUSION: IR-780 possessed the ability to visualize a variety of ccRCC cells, but not for normal renal epithelial cells. IR-780 had obvious overlapping staining in lysosomes or mitochondria, indicating its selective accumulation in mitochondria and lysosomes in the organelles. IR-780 could detect a small amount of ccRCC in the blood. These results suggest that the near-infrared fluorescent dye IR-780 can specifically accumulate in ccRCC cells, which can be used for specific diagnosis of ccRCC.
引文
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[14]Siegel RL,Miller KD,Jemal A.Cancer statistics,2016.CA Cancer JClin.2016;66(1):7-30.
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[18]Tan HJ,Norton EC,Ye Z,et al.Long-term survival following partial vs radical nephrectomy among older patients with early-stage kidney cancer.JAMA.2012;307(15):1629-1635.
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[20]Yang X,Shi C,Tong R,et al.Near IR heptamethine cyanine dye-mediated cancer imaging.Clin Cancer Res.2010;16(10):2833-2844.
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[22]Shi C,Zhang C,Su Y,et al.Cyanine dyes in optical imaging of tumours.Lancet Oncol.2010;11(9):815-816.
[23]Zhang C,Wang S,Xiao J,et al.Sentinel lymph node mapping by a near-infrared fluorescent heptamethine dye.Biomaterials.2010;31(7):1911-1917.
[24]田迎,卢光明,郑玲近红外荧光染料用于肿瘤检测的初步研究[J].医疗卫生装备,2015(10):24-26.
[25]Zhang E,Luo S,Tan X,et al.Mechanistic study of IR-780 dye as a potential tumor targeting and drug delivery agent.Biomaterials.2014;35(2):771-778.
[26]Yi X,Zhang J,Yan F,et al.Synthesis of IR-780 dye-conjugated abiraterone for prostate cancer imaging and therapy.Int J Oncol.2016;49(5):1911-1920.
[27]Kawaguchi M,Hara N,Bilim V,et al.A diagnostic marker for superficial urothelial bladder carcinoma:lack of nuclear ATBF1(ZFHX3)by immunohistochemistry suggests malignant progression.BMC Cancer.2016;16(1):805.
[28]Wu JB,Shao C,Li X,et al.Near-infrared fluorescence imaging of cancer mediated by tumor hypoxia and HIF1alpha/OATPs signaling axis.Biomaterials.2014;35(28):8175-8185.
[29]Tian Y,Sun J,Yan H,et al.A rapid and convenient method for detecting a broad spectrum of malignant cells from malignant pleuroperitoneal effusion of patients using a multifunctional NIR heptamethine dye.Analyst.2015;140(3):750-755.
[2]Fitzmaurice C,Dicker D,Pain A,et al.The Global Burden of Cancer2013.JAMA Oncol.2015;1:505-527.
[3]Miller KD,Siegel RL,Lin CC,et al.Cancer treatment and survivorship statistics,2016.CA Cancer J Clin.2016;66:271-289.
[4]Siegel RL,Miller KD,Jemal A.Cancer Statistics,2017.CA Cancer J Clin.2017;67:7-30.
[5]Prokofiev D,Kreutzer N,Kress A,et al.Small renal mass.Urologe A.2012;51(10):1459-1465,1466-1468.
[6]Ficarra V,Bhayani S,Porter J,et al.Predictors of warm ischemia time and perioperative complications in a multicenter,international series of robot-assisted partial nephrectomy.Eur Urol.2012;61(2):395-402.
[7]Ljungberg B,Bensalah K,Canfield S,et al.EAU guidelines on renal cell carcinoma:2014 update.Eur Urol.2015;67(5):913-924.
[8]Biffi S,Garrovo G,Maeor P,et al.In vivo biodistribution and lifetime analysis of Cy5.5-connjugated ritaximab in mice bearing lymphoid tumor xenograft using time-domain near-infrared optical imaging.Mol Imaging.2008;7(6):272-282.
[9]Medarova Z,Rashkovetsky L,Pantaztpoulos P,et al.Multiparametric monitoring of tumor response to chemotherapy by noninvasive imaging.Cancer Res.2009;69(3):1182-1189.
[10]Yi X,Wang F,Qin W,et al.Near-infrared fluorescent probes in cancer imaging and therapy:an emerging field.Int J Nanomedicine.2014;9:1347-1365.
[11]Guo Z,Park S,Yoon J,et al.Recent progress in the development of near-infrared fluorescent probes for bioimaging applications.Chem Soc Rev.2014;43(1):16-29.
[12]Hilderbrand SA,Weissleder R.Near-infrared fluorescence:application to in vivo molecular imaging.Curr Opin Chem Biol.2010;14(1):71-79.
[13]Yang X,Shao C,Wang R,et al.Optical imaging of kidney cancer with novel near infrared heptamethine carbocyanine fluorescent dyes.JUrol.2013;189(2):702-710.
[14]Siegel RL,Miller KD,Jemal A.Cancer statistics,2016.CA Cancer JClin.2016;66(1):7-30.
[15]Chen W,Zheng R,Zeng H,et al.The incidence and mortality of major cancers in China,2012.Chin J Cancer.2016;35(1):73.
[16]叶章群,孙颖浩,那彦群,等.中国泌尿外科疾病诊断治疗指南[M].北京:人民卫生出版社,2014.
[17]Caputo PA,Ramirez D,Zargar H,et al.Laparoscopic Cryoablation for Renal Cell Carcinoma:100-Month Oncologic Outcomes.J Urol.2015;194(4):892-896.
[18]Tan HJ,Norton EC,Ye Z,et al.Long-term survival following partial vs radical nephrectomy among older patients with early-stage kidney cancer.JAMA.2012;307(15):1629-1635.
[19]Hilderbrand SA,Weissleder R.Near-infrared fluorescence:application to in vivo molecular imaging.Curr Opin Chem Biol.2010;14(1):71-79.
[20]Yang X,Shi C,Tong R,et al.Near IR heptamethine cyanine dye-mediated cancer imaging.Clin Cancer Res.2010;16(10):2833-2844.
[21]贺丽芳,黄文河,张国君.恶性肿瘤的光学分子影像手术导航[J].中国肿瘤临床,2016,43(21):927-931.
[22]Shi C,Zhang C,Su Y,et al.Cyanine dyes in optical imaging of tumours.Lancet Oncol.2010;11(9):815-816.
[23]Zhang C,Wang S,Xiao J,et al.Sentinel lymph node mapping by a near-infrared fluorescent heptamethine dye.Biomaterials.2010;31(7):1911-1917.
[24]田迎,卢光明,郑玲近红外荧光染料用于肿瘤检测的初步研究[J].医疗卫生装备,2015(10):24-26.
[25]Zhang E,Luo S,Tan X,et al.Mechanistic study of IR-780 dye as a potential tumor targeting and drug delivery agent.Biomaterials.2014;35(2):771-778.
[26]Yi X,Zhang J,Yan F,et al.Synthesis of IR-780 dye-conjugated abiraterone for prostate cancer imaging and therapy.Int J Oncol.2016;49(5):1911-1920.
[27]Kawaguchi M,Hara N,Bilim V,et al.A diagnostic marker for superficial urothelial bladder carcinoma:lack of nuclear ATBF1(ZFHX3)by immunohistochemistry suggests malignant progression.BMC Cancer.2016;16(1):805.
[28]Wu JB,Shao C,Li X,et al.Near-infrared fluorescence imaging of cancer mediated by tumor hypoxia and HIF1alpha/OATPs signaling axis.Biomaterials.2014;35(28):8175-8185.
[29]Tian Y,Sun J,Yan H,et al.A rapid and convenient method for detecting a broad spectrum of malignant cells from malignant pleuroperitoneal effusion of patients using a multifunctional NIR heptamethine dye.Analyst.2015;140(3):750-755.