RESUMO
The modification of EGFR aptamer (Apt 1) and HER2 aptamer (Apt 2) with gold nanoparticles (AuNPs) is reported to obtain probe I (Apt 1-AuNPs) and probe II (Apt 2-AuNPs). Taking Eca109, KYSE510, and KYSE150 cells as models, the sandwich scattering system of probe I-cell-probe II was formed by the recognition of tumor markers by the aptamer modified probe, and the resonance Rayleigh scattering (RRS) spectra were investigated. The results showed that the scattering system can be used to quantitatively detect the Eca109 cell lines in the range 5.0×10 to 5.0×105 cells·mL-1 with a detection limit of 15 cells· mL-1.The system can also detect the KYSE510 cell lines in a linear range of 5.0×10 to 5.0×105 cells·mL-1 with a detection limit of 18 cells·mL-1 and the KYSE150 cell lines in a linear range of 3.0×10 to 5.0×105 cells·mL-1 with a detection limit of 12 cells·mL-1. To demonstrate the potential application of the RRS method for real sample analysis, cells were spiked into blank serum samples at concentrations from 1.0×102 to 1.0×105 cells·mL-1. The recovery was between 97.0% and 102.3%, and the RSD was between 1.1% and 4.9%, confirming the feasibility of the proposed method for ESCC cell determination.
Assuntos
Aptâmeros de Nucleotídeos , Neoplasias Esofágicas , Nanopartículas Metálicas , Humanos , Ouro , Neoplasias Esofágicas/diagnósticoRESUMO
Telomerase is considered a valuable diagnostic and prognostic cancer biomarker. Accurate and reliable detection of telomerase activity is of great value in clinical diagnosis, screening of inhibitors, and therapeutics. Here, we developed a novel amplified fluorescence resonance energy transfer (FRET) nanoprobe for highly sensitive and reliable monitoring of intracellular telomerase activity. The nanoprobe (QDSA@DNA) was composed of a streptavidin-modified quantum dot (QDSA) which was functionalized with a telomerase primer sequence (TP) and Cy5-tagged signal switching sequence (SS) through biotin-streptavidin interaction. When the nanoprobe was assembled, the Cy5 was in close proximity to the QDSA, resulting in high FRET efficiency from the QDSA to Cy5. In the presence of telomerase, the TP could be extended to produce telomeric repeat units, which was complementary to the loop of SS. Thus, the SS could hybridize with elongated sequences to form a rigid double-stranded structure, which forced the Cy5 away from the surface of the QDSA, causing low FRET efficiency. Furthermore, due to the production of multiple repeat units by telomerase, multiple hairpin structures could be opened, yielding significant fluorescence ratio (FQDsa/FCy5) enhancement for sensing of telomerase activity. In this way, the combination of a FRET and target-assisted strategy in a nanoprobe improved the detection accuracy and amplified the detection signal, respectively. The QDSA@DNA nanoprobe also showed high selectivity, excellent nuclease stability, and good biocompatibility. More importantly, this nanoprobe was found to be an excellent platform for efficient monitoring of intracellular telomerase activity, providing a potential platform in tumor diagnosis and screening of telomerase-related inhibitors.
Assuntos
Corantes Fluorescentes/química , Nanoestruturas/química , Telomerase/metabolismo , Transferência Ressonante de Energia de Fluorescência/métodos , Células HeLa , Humanos , Pontos QuânticosRESUMO
Although great achievements have been made in the synthesis of giant lanthanide clusters, novel structural models are still scarce. Herein, we report a giant lanthanide cluster Dy76 , constructed from [Dy3 (µ3 -OH)4 ] and [Dy5 (µ4 -O)(µ3 -OH)8 ] building blocks. As the largest known Dy cluster, the structure of Dy76 can be seen as arising from the fusion of two Dy48 clusters; these clusters can be isolated under various synthetic conditions and were characterized by single-crystal X-ray diffraction. This new, fused structural model of the pillar motif has not been found in Ln clusters. Furthermore, the successful conversion of Dy76 back into Dy48 in a retrosynthetic manner supports the proposed fusion formation mechanism of Dy76 . Electrospray ionization mass spectrometry (ESI-MS) analysis suggests that the metal cluster skeleton of Dy76 shows good stability in various solvents. This work not only reveals a new structural type of Ln clusters but also provides insight into the novel fusion assembly process.