利用生物纳米技术检测外周血肺癌细胞的实验研究

目的建立磁性纳米颗粒从肿瘤患者外周血中富集与纯化肿瘤细胞的方法，应用荧光纳米晶生物探针检测方法，实现对肺癌早期微转移的诊断。方法体外培养人肝癌细胞和肺癌细胞，采集健康人外周血单个核细胞（PBMC），将PBMC与肝癌细胞和肺癌细胞按比例混合，通过磁粒抗体复合物富集癌细胞，用荧光纳米晶探针进行肺癌特异性鉴定。结果光镜下可见较多的人肝癌细胞和人肺癌细胞与磁性纳米颗粒紧密结合，而PBMC与磁性纳米颗粒不结合。荧光显微镜下可见荧光纳米晶表面蛋白A探针特异性地与肺癌细胞结合产生荧光信号，但与肝癌细胞不结合，无荧光信号出现。结论本方法可成功地从外周血中富集到肺癌细胞，经荧光纳米晶探针成功地鉴定为肺癌细胞，达到诊断肺癌早期微转移的目的。

Detection of A549 pulmonary carcinoma cells by bio-nanotechnology

Objective To establish a method of enriching tumor cells from peripheral blood of tumor patients by superparamagnetic nanoparticles, and using fluorescence nanocrystals bioprobe method to realize no injury diagnosis-to earlier period pulmonary carcinoma micrometastasis. Methods Hepatoma carcinoma cells and pulmonary carcinoma cells were cultured in vitro, Mononuclear cells（PBMCs） in healthy person＇s peripheral blood were collected. PBMCs were mixed with liver cancer and lung cancer ceils by proportion, and enriched tumor ceils by superparamagnetic nanoparticle antibody com- plex, lung cancer specificness was identified by fluorescence nanocrystais bioprobe. We smeared tumor cells on glass slide and identified pulmonary carcinoma cells specificity by fluorescence nanocrystals bioprobe. Results We could find many hepatoma carcinoma ceils and pulmonary carcinoma cells binding tightly with magnetic nanoparticles under the light micro-scope, but at the same conditions, PBMC couldn＇t bind with magnetic nanoparticles. Nanocrystals SP-A bioprebe could specially bind with pulmonary carcinoma cells, and produced fluorescence signal, but did not bind with hepatoma carcinoma cell, and no fluorescence signal appeared. Conclusions This method can enrich tumor cells from peripheral blood of tumor patients SP-A bioprobe, and the lung cancer ceils are identified successfully by fluorescence nanocrystals bioprobe, and achieved the purpose of diagnosis of earlier micrometastasis of lung cancer.