CIGMH, Departamento de Ciências da Vida, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Campus de Caparica, 2829-516 Caparica, Portugal.
J Nanobiotechnology. 2010 Feb 24;8:5. doi: 10.1186/1477-3155-8-5.
Molecular nanodiagnostics applied to cancer may provide rapid and sensitive detection of cancer related molecular alterations, which would enable early detection even when those alterations occur only in a small percentage of cells. The use of gold nanoparticles derivatized with thiol modified oligonucleotides (Au-nanoprobes) for the detection of specific nucleic acid targets has been gaining momentum as an alternative to more traditional methodologies. Here, we present an Au-nanoparticles based approach for the molecular recognition and quantification of the BCR-ABL fusion transcript (mRNA), which is responsible for chronic myeloid leukemia (CML), and to the best of our knowledge it is the first time quantification of a specific mRNA directly in cancer cells is reported. This inexpensive and very easy to perform Au-nanoprobe based method allows quantification of unamplified total human RNA and specific detection of the oncogene transcript. The sensitivity settled by the Au-nanoprobes allows differential gene expression from 10 ng/mul of total RNA and takes less than 30 min to complete after total RNA extraction, minimizing RNA degradation. Also, at later stages, accumulation of malignant mutations may lead to resistance to chemotherapy and consequently poor outcome. Such a method, allowing for fast and direct detection and quantification of the chimeric BCR-ABL mRNA, could speed up diagnostics and, if appropriate, revision of therapy. This assay may constitute a promising tool in early diagnosis of CML and could easily be extended to further target genes with proven involvement in cancer development.
应用于癌症的分子纳米诊断技术可以快速、灵敏地检测到与癌症相关的分子变化，即使这些变化仅发生在一小部分细胞中，也能实现早期检测。用巯基修饰的寡核苷酸衍生的金纳米粒子（Au-纳米探针）来检测特定的核酸靶标，已经作为一种替代更传统方法的方法得到了越来越多的应用。在这里，我们提出了一种基于 Au 纳米粒子的方法，用于分子识别和定量检测导致慢性髓性白血病（CML）的 BCR-ABL 融合转录本（mRNA），据我们所知，这是首次直接在癌细胞中定量检测特定 mRNA。这种廉价且非常易于操作的基于 Au 纳米探针的方法可以定量未扩增的总人类 RNA，并特异性检测致癌基因转录本。Au 纳米探针的灵敏度可以从 10ng/mul 的总 RNA 中检测到差异基因表达，并且在总 RNA 提取后不到 30 分钟即可完成，最大限度地减少了 RNA 降解。此外，在后期，恶性突变的积累可能导致对化疗的耐药性，从而导致预后不良。这种能够快速、直接检测和定量嵌合 BCR-ABL mRNA 的方法，可以加快诊断速度，如果需要，还可以修改治疗方案。该检测方法有望成为 CML 早期诊断的一种有前途的工具，并且可以很容易地扩展到进一步具有明确参与癌症发展的靶基因。
