State Key Laboratory of Natural and Biomimetic Drugs, Beijing Key Laboratory of Molecular Pharmaceutics and New Drug Delivery Systems, and Department of Medicinal Chemistry, School of Pharmaceutical Sciences , Peking University , No. 38, Xueyuan Road , Beijing 100191 , P.R. China.
Anal Chem. 2019 Sep 3;91(17):11045-11054. doi: 10.1021/acs.analchem.9b01382. Epub 2019 Aug 13.
Early precise diagnosis of cancers is crucial to realize more effective therapeutic interventions with minimal toxic effects. Cancer phenotypes may also alter greatly among patients and within individuals over the therapeutic process. The identification and characterization of specific biomarkers expressed on tumor cells are in high demand for diagnosis and treatment, but they are still a challenge. Herein, we designed three new bioorthogonal surface-enhanced Raman scattering (SERS) nanoprobes and successfully applied the cocktail of them for MDA-MB-231 and MCF-7 breast cancer multiplex phenotype detection. The SERS nanoprobes containing Raman reporters with diynl, azide, or cyano moieties demonstrated apparent Raman shift peaks in 2205, 2120, and 2230 cm, respectively, in the biologically Raman-silent region. Three target ligands, including oligonucleotide aptamer (AS1411), arginine-glycine-aspatic acid (RGD) peptide, and homing cell adhesion molecule antibody (anti-CD44), were separately conjugated to the nanoprobes for active recognition capability. The cocktail of the nanoprobes manifested minimal cytotoxicity and simultaneously multiplex phenotype imaging of MDA-MB-231 and MCF-7 cells. Quantitative measurement of cellular uptake by inductively coupled plasma mass spectrometry (ICPMS) verified that MDA-MB-231 cells harbored a much higher expression level of CD44 receptor than MCF-7 cells. For in vivo SERS detection, Raman shift peaks of 2120, 2205, and 2230 cm in the micro-tumor were clearly observed, representing the existence of three specific biomarkers of nucleolin, integrin αβ, and CD44 reporter, which could be used for early cancer phenotype identification. The biodistribution results indicated that target ligand modified nanoprobes exhibited much more accumulation in tumors than those nanoprobes without target ligands. The multicolor cocktail of bioorthogonal SERS nanoprobes offers an attractive and insightful strategy for early cancer multiplex phenotype targeting and diagnosis in vivo that is noninvasive and has low cross-talk, unique spectral-molecular signature, high sensitivity, and negligible background interference.
早期精确诊断癌症对于实现更有效的治疗干预措施并将毒副作用降至最低至关重要。癌症表型在患者之间以及治疗过程中个体内也可能发生很大变化。因此，鉴定和表征肿瘤细胞上表达的特定生物标志物对于诊断和治疗非常重要，但这仍然是一个挑战。在此，我们设计了三种新的生物正交表面增强拉曼散射（SERS）纳米探针，并成功地将它们的混合物用于 MDA-MB-231 和 MCF-7 乳腺癌多重表型检测。含有二炔、叠氮或氰基部分的拉曼报告分子的 SERS 纳米探针在生物拉曼静默区分别显示出 2205、2120 和 2230cm 的明显拉曼位移峰。三种靶配体，包括寡核苷酸适体（AS1411）、精氨酸-甘氨酸-天冬氨酸（RGD）肽和归巢细胞黏附分子抗体（抗-CD44），分别被连接到纳米探针对其进行主动识别能力。纳米探针的混合物表现出最小的细胞毒性，并同时对 MDA-MB-231 和 MCF-7 细胞进行多重表型成像。通过电感耦合等离子体质谱（ICPMS）进行的细胞摄取定量测量证实，MDA-MB-231 细胞的 CD44 受体表达水平明显高于 MCF-7 细胞。用于体内 SERS 检测，微肿瘤中 2120、2205 和 2230cm 的拉曼位移峰清晰可见，代表核仁素、整合素 αβ 和 CD44 报告物的三种特定生物标志物的存在，可用于早期癌症表型鉴定。生物分布结果表明，与没有靶配体的纳米探针相比，靶配体修饰的纳米探针在肿瘤中的积累更多。生物正交 SERS 纳米探针的多色混合物为体内早期癌症多重表型靶向和诊断提供了一种有吸引力和有见地的策略，具有非侵入性、低串扰、独特的光谱-分子特征、高灵敏度和可忽略的背景干扰。
