Int J Nanomedicine 2011 ;6 2175-81. [IF:4.976]
Detection of micrometastases in peripheral blood of non-small cell lung cancer with a refined immunomagnetic nanoparticle enrichment assay.
Li Q , Qi H , Zhou HX , Deng CY , Zhu H , Li JF , Wang XL , Li FR .
Clinical Medical Research Center, The Second Clinical Medical College (Shenzhen People's Hospital), Jinan University, Shenzhen, People's Republic of China.
深圳市人民医院临床医学研究中心，暨南大学
Abstract
Fe(3)O(4) 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 μg/mg. We could detect down to one tumor cell in 5 × 10(7) 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.
摘要:
目前Fe(3)O(4)颗粒作为免疫磁性微球核心用于血循环中肿瘤细胞（CTCs）的免疫磁性富集分析。现在很难进一步改进CTC检测的敏感性以及对特定肿瘤类型、特征的辨认。在本研究中，我们制备了以纳米铁为核心，外面包被抗细胞角蛋白7/8(CK7/8)单克隆抗体的免疫磁性纳米颗粒。该免疫磁性纳米颗粒联合免疫细胞化学(ICC)即免疫磁性纳米颗粒富集分析用于检测非小细胞肺癌CTC。对比该分析方法与巢式逆转录多聚酶链反应检测（巢式RT-PCR）CK19mRNA和肺特异性X蛋白(LUNX)mRNA的结果；以人肺腺癌细胞A549评价检测方法的敏感性和特异性；抽取外周血检测CTC。免疫磁性纳米颗粒的平均直径为51nm，吸附的抗体量为111.2μg/mg。只要5*107个外周血单核细胞中有一个肿瘤细胞就能检测到。结果表明该方法和巢式RT-PCR敏感性类似，但假阳性显著降低。改良的免疫磁性分析联合ICC与巢式RT-PCR相比，敏感性相似，但特异性显著提高。综上所述，该技术能够用于检测微转移，重新定义临床分期以及指导个体化的术后治疗；该技术有重要的临床价值需行进一步的临床试验。
