AFP-GP73-miRNA多效纳米探针的构建及其在肝癌诊断中的应用研究

It's lack accuracy under the commonly employed alpha-fetal protein (AFP) detection in serum for hepatocellular carcinoma (HCC). Reports show that combined detection with multi-biomarker would improve the diagnosis of cancers. Therefore, this project will firstly construct two kinds nano probes with the core of CdTe quantum dot and magnetic iron oxide separately, combine AFP with the novel biomarker, Golgi protein-73 (GP73) and also with newly founded miR-500 and miR-21, in order to increase the sensitivity and specificity of HCC detection. Then, clinical plasm of HCC will be detected to explore a novel non-involved early diagnosis method. Meanwhile, with the unique advantages of nanoparticles like having high surface area, fluorescence property, magnetic character and so on, thus easily to achieve the goals of multiple effects within one step operation. Besides, the projece will compare these two nano probe and figure out their value in HCC diagnosis, and to evaluate their potential using in detection devices. This research is aiming to explore the non-invasion diagnosis and future integration detection of HCC through plasm, and will play an important role in improving the early detection, cancer diagnosis level and treatment of HCC.

临床常用的肝癌标志物如甲胎蛋白（AFP）尚不理想，对于肝癌的早期诊断意义不大。临床实践说明，肿瘤标志物联检方法可提高肿瘤的诊断水平。为此，本项目拟先以荧光碲化镉量子点或磁性四氧化三铁纳米材料为载体，负载AFP、肝癌新标志物高尔基体蛋白（GP73）、致癌性微小RNA miR-500和miR-21等肝癌标志物，然后结合临床标本的检测希望建立一种新颖的非创性肝癌早期诊断新方法。纳米材料不仅比表面积大，易富集多种标志物；而且具有荧光、磁性等特性，可方便仪器检测，如用于检测血清标志物将可能实现一次检测体现多项肿瘤标志物数值的目标。本项目还要比较这两种纳米探针在肝癌诊断中的优缺点，评价纳米光/磁特性在肝癌血清检测中的应用价值及进行非侵入式肝癌早期检测器件开发的潜力，不但在血清检测领域中具有较好的探索性，而且将有利于提高肝癌早期诊断的水平，对于提高我国肝癌的诊治水平起积极的推动作用。

早期诊断对肝癌来说仍存在挑战。目前临床常用的肝癌标志物如甲胎蛋白（AFP）尚不理想。于是我们采用组合思路设计构建纳米复合材料，对病人血样进行体外非侵入的检查，目的是探索纳米复合探针在肝细胞肝癌（HCC）早期诊断中的可行性。我们的研究中成功得到可控碲化镉纳米颗粒，成功体现了尺寸与发射光谱的相关性。并且评价了细胞毒性。也在细胞水平实验中得到预期标记效果。在与APF结合后体现了纳米探针的功效。这些结果提示其标志探针性质是具有进一步优化和推广应用价值的。在肝癌细胞模型以及真实肝癌患者血清检测中，我们的探针体现了更高的灵敏性。这些结果极具开发和经济价值。在多种蛋白复合处理时候没有得到预期结果。蛋白的稳定性、摩尔比、活性、及产率都在可控范围之外，重复性不理想。