Detection of micrometastases in peripheral blood of non-small cell lung cancer with a refined immunomagnetic nanoparticle enrichment assay

Qing Li1, Hui Qi1, Han-Xin Zhou1, Chun-Yan Deng1, Hai Zhu3, Jin-Feng Li3, Xi-Li Wang3, Fu-Rong Li1,21Clinical Medical Research Center, The Second Clinical Medical College (Shenzhen People&#39;s Hospital), Jinan University, Shenzhen, People&#39;s Republic of China; 2Shenzhen Institute...

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Autores principales:	Li Q, Qi H, Zhou HX, Deng CY, Zhu H, Li JF, Wang XL, Li FR
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2011
Materias:	Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/cf71e88965614b4fa433331e2d25932e
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spelling	oai:doaj.org-article:cf71e88965614b4fa433331e2d25932e2021-12-02T02:14:38ZDetection of micrometastases in peripheral blood of non-small cell lung cancer with a refined immunomagnetic nanoparticle enrichment assay1176-91141178-2013https://doaj.org/article/cf71e88965614b4fa433331e2d25932e2011-09-01T00:00:00Zhttp://www.dovepress.com/detection-of-micrometastases-in-peripheral-blood-of-non-small-cell-lun-a8384https://doaj.org/toc/1176-9114https://doaj.org/toc/1178-2013Qing Li1, Hui Qi1, Han-Xin Zhou1, Chun-Yan Deng1, Hai Zhu3, Jin-Feng Li3, Xi-Li Wang3, Fu-Rong Li1,21Clinical Medical Research Center, The Second Clinical Medical College (Shenzhen People&#39;s Hospital), Jinan University, Shenzhen, People&#39;s Republic of China; 2Shenzhen Institute of Gerontology, Shenzhen, People&#39;s Republic of China; 3Shenzhen Bioeasy Biotechnologies Co Ltd, Shenzhen, People&#39;s Republic of ChinaAbstract: Fe3O4 particles are currently used as the core of immunomagnetic microspheres in the immunomagnetic enrichment assay of circulating tumor cells (CTCs). It is difficult to further improve the sensitivity of CTC detection or to improve tumor cell-type identification and characterization. In the present study, we prepared immunomagnetic nanoparticles with nanopure iron as the core, coated with anti-cytokeratin 7/8 (CK7/8) monoclonal antibody. These immunomagnetic nanoparticles (IMPs) were used in conjunction with immunocytochemistry (ICC) to establish a refined immunomagnetic nanoparticle enrichment assay for CTC detection in non-small cell lung cancer (NSCLC). The assay was compared with nested reverse transcription polymerase chain reaction (RT-PCR) to detect CK19 mRNA and lung specific X protein (LUNX) mRNA. Human lung adenocarcinoma cell line A549 was used for sensitivity and specificity evaluation. Peripheral blood samples were collected from each group for CTC detection. The average diameter of the immunomagnetic nanoparticles was 51 nm, and the amount of adsorbed antibodies was 111.2 &micro;g/mg. We could detect down to one tumor cell in 5 &times; 107 peripheral blood mononuclear cells. The sensitivity was consistent with that of nested RT-PCR; however, the false positive rate was significantly reduced. The modified assay combined with ICC did not differ from nested RT-PCR in sensitivity, but it had significantly increased specificity. This approach could, therefore, contribute to identification of micrometastases, re-defining clinical staging, and guiding individual postoperative treatments. The technique shows considerable potential clinical value and further clinical trials are warranted.Keywords: NSCLC, circulating tumor cells, nested RT-PCR, immunomagnetic nanoparticles, immunocytochemistryLi QQi HZhou HXDeng CYZhu HLi JFWang XLLi FRDove Medical PressarticleMedicine (General)R5-920ENInternational Journal of Nanomedicine, Vol 2011, Iss default, Pp 2175-2181 (2011)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine (General) R5-920
spellingShingle	Medicine (General) R5-920 Li Q Qi H Zhou HX Deng CY Zhu H Li JF Wang XL Li FR Detection of micrometastases in peripheral blood of non-small cell lung cancer with a refined immunomagnetic nanoparticle enrichment assay
description	Qing Li1, Hui Qi1, Han-Xin Zhou1, Chun-Yan Deng1, Hai Zhu3, Jin-Feng Li3, Xi-Li Wang3, Fu-Rong Li1,21Clinical Medical Research Center, The Second Clinical Medical College (Shenzhen People&#39;s Hospital), Jinan University, Shenzhen, People&#39;s Republic of China; 2Shenzhen Institute of Gerontology, Shenzhen, People&#39;s Republic of China; 3Shenzhen Bioeasy Biotechnologies Co Ltd, Shenzhen, People&#39;s Republic of ChinaAbstract: Fe3O4 particles are currently used as the core of immunomagnetic microspheres in the immunomagnetic enrichment assay of circulating tumor cells (CTCs). It is difficult to further improve the sensitivity of CTC detection or to improve tumor cell-type identification and characterization. In the present study, we prepared immunomagnetic nanoparticles with nanopure iron as the core, coated with anti-cytokeratin 7/8 (CK7/8) monoclonal antibody. These immunomagnetic nanoparticles (IMPs) were used in conjunction with immunocytochemistry (ICC) to establish a refined immunomagnetic nanoparticle enrichment assay for CTC detection in non-small cell lung cancer (NSCLC). The assay was compared with nested reverse transcription polymerase chain reaction (RT-PCR) to detect CK19 mRNA and lung specific X protein (LUNX) mRNA. Human lung adenocarcinoma cell line A549 was used for sensitivity and specificity evaluation. Peripheral blood samples were collected from each group for CTC detection. The average diameter of the immunomagnetic nanoparticles was 51 nm, and the amount of adsorbed antibodies was 111.2 &micro;g/mg. We could detect down to one tumor cell in 5 &times; 107 peripheral blood mononuclear cells. The sensitivity was consistent with that of nested RT-PCR; however, the false positive rate was significantly reduced. The modified assay combined with ICC did not differ from nested RT-PCR in sensitivity, but it had significantly increased specificity. This approach could, therefore, contribute to identification of micrometastases, re-defining clinical staging, and guiding individual postoperative treatments. The technique shows considerable potential clinical value and further clinical trials are warranted.Keywords: NSCLC, circulating tumor cells, nested RT-PCR, immunomagnetic nanoparticles, immunocytochemistry
format	article
author	Li Q Qi H Zhou HX Deng CY Zhu H Li JF Wang XL Li FR
author_facet	Li Q Qi H Zhou HX Deng CY Zhu H Li JF Wang XL Li FR
author_sort	Li Q
title	Detection of micrometastases in peripheral blood of non-small cell lung cancer with a refined immunomagnetic nanoparticle enrichment assay
title_short	Detection of micrometastases in peripheral blood of non-small cell lung cancer with a refined immunomagnetic nanoparticle enrichment assay
title_full	Detection of micrometastases in peripheral blood of non-small cell lung cancer with a refined immunomagnetic nanoparticle enrichment assay
title_fullStr	Detection of micrometastases in peripheral blood of non-small cell lung cancer with a refined immunomagnetic nanoparticle enrichment assay
title_full_unstemmed	Detection of micrometastases in peripheral blood of non-small cell lung cancer with a refined immunomagnetic nanoparticle enrichment assay
title_sort	detection of micrometastases in peripheral blood of non-small cell lung cancer with a refined immunomagnetic nanoparticle enrichment assay
publisher	Dove Medical Press
publishDate	2011
url	https://doaj.org/article/cf71e88965614b4fa433331e2d25932e
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Detection of micrometastases in peripheral blood of non-small cell lung cancer with a refined immunomagnetic nanoparticle enrichment assay

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