Cytochrome c (cyt c) and caspase-9 were critical biomarkers in mitochondria-mediated apoptosis. A novel electrochemical immunosensor was developed for in situ analysis of cyt c and caspase-9 in the cytosol. Gold nanoparticle-polydopamine (AuNP/PDA) composites were used to fabricate the interface of the sensor. The anti-cyt c or anti-caspase-9 functionalized-immunosensor provided a biomimetic interface for immunosensing of cyt c or caspase-9 in Hela cells during apoptosis. The changes in the expression level of cyt c and caspase-9 in the cytosol upon curcumin-induced apoptosis were detected by using the proposed method, and also the influence of different concentrations and incubation times of curcumin-induced Hela cells was investigated. This method achieved a linear range (0.1-100 μM) for standard cyt c and caspase-9, with a detection limit of 0.03 ± 0.01 μM for standard cyt c and 0.08 ± 0.02 μM for standard caspase-9. Moreover, this method was used to detect cells which could detect as low as 100 cells which expressed cyt c and caspase-9, and also the results are in good agreement with standard flow cytometry analysis. The developed electrochemical immunosensor offered a simple and rapid approach for sensitive evaluation of apoptosis markers with considerable specificity and reproducibility, and also the developed strategy could be of great importance in clinical diagnosis and therapeutic research.
A simple method for preparing quercetin surface-functionalized germanium nanoparticles (Qu-GeNPs) with enhanced antioxidant and anticancer activity is reported. Spherical germanium nanoparticles (GeNPs) were capped by quercetin (Qu) with a mean particle size of approximately 33 nm and were characterized by TEM, AFM, UV-visible absorption spectroscopy, FTIR, and XRD measurements. The in vitro drug release of Qu from the Qu-GeNPs indicated that Qu could principally be distributed around tumor tissues rather than in the normal section and Qu-GeNPs were internalized by MCF-7 cells. Their biological activity test results indicated that these Qu-GeNPs possessed stronger hydroxyl-scavenging effects and proliferative inhibition effect on MCF-7 cancer cells than quercetin, thus suggesting that the strategy to use GeNPs as a carrier of Qu could be an efficient way to achieve enhanced antioxidant and anticancer activity. In addition, Qu-GeNPs possessed a high apoptotic induction effect in cancer cells, especially in high dosages, and could arrest MCF-7 cells in the S phase.
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