Developing novel instruments and technologies for spatio-temporal and dynamic meas-urements of the intricate cellular effects involving molecular translocation, signal transduction, and molecular interactions inside l...Developing novel instruments and technologies for spatio-temporal and dynamic meas-urements of the intricate cellular effects involving molecular translocation, signal transduction, and molecular interactions inside living cells is essential for the cell and molecular biology science. For the purpose of monitoring and investigating molecular events in living cells at real-time, the surface plasmon resonance based cytosensor (SBCS) for cell cultur-ing and signal monitoring was established, and on the basis of it, the corresponding technology was also established by monitoring and analyzing SPR re-sponses induced in rat C6 glioma cells by phorbol 12-myristate 13-acetate (PMA). The SPR signals induced by PMA in living C6 cells were significantly different from those groups without cells. These re-sponses were strongly dependent on and saturable to the concentrations of PMA, and could be sup-pressed by the specific and potent PKC inhibitors, which indicated that the measured signal could be the reflection of the redistribution of intracellular components near the cell membrane triggered by the activation of PKC. This research provides a quantita-tive and non-invasive technique to study the spa-tio-temporal characteristics of the cellular effects in living cells at real-time. Furthermore, this technology could also be widely used in the basic research as well as applied realms, such as space effects evaluation, environmental safety assessment, bio-logical weapon detection, cellular and molecular re-search, and drug screening.展开更多
Circulating tumor cells(CTCs)are important markers for cancer.The part of tumor cells that are detached from the primary tumor or metastatic tumor and enter the blood circulation is called CTCs.It is crucial to develo...Circulating tumor cells(CTCs)are important markers for cancer.The part of tumor cells that are detached from the primary tumor or metastatic tumor and enter the blood circulation is called CTCs.It is crucial to develop a rapid,accurate,and easy-to-implement diagnostic system for CTCs for early tumor diagnosis and for monitoring progression of the disease.In this work,we reported a colorimetric and fluorescent dual-mode assay for the detection of CTCs.Our assay used magnetic nanoparticles and aptamer for CTCs capture and gold nanoparticles-loaded covalent organic frameworks(Au@COFs)for signal amplification,respectively.The magnetic nanoparticles were modified with folic acid to capture CTCs by interaction between folic acid and the folate receptor overexpressed on the surface of tumor cells.The covalent organic frameworks were engineered to have both nitro-reductase-like and glucose–oxidase-like activities.The nitro-reductase-like activity converted the substrate p-nitrophenol to p-nitroaniline for colorimetric detection,and the glucose-oxidase-like activity enabled fluorescence detection.Specifically,Au@COFs catalyzed glucose oxidation and generated hydrogen peroxide to oxidize Fe 2+to Fe 3+,which converted MIL(Al)-MOF to MIL(Fe)-MOF through ion exchange,resulting in the fluorescence quenching of MIL(Al)-MOF.Our assay showed high sensitivity with a detection limit of 17 cells/mL using MCF-7 cells as model cancer cells.This work provided an e fficient and ultrasensitive strategy for CTCs detection and has potential applications in cancer identification and diagnosis.Dual-mode detection system,combining colorimetric and fluorescent signals,integration of magnetic nanoparticles and aptamers for CTC capture,and utilization of covalent organic frameworks loaded with gold nanoparticles for signal amplification can improve sensitivity and accuracy.展开更多
基金supported by the National Natural Science Foundation(Grant No.30370365).
文摘Developing novel instruments and technologies for spatio-temporal and dynamic meas-urements of the intricate cellular effects involving molecular translocation, signal transduction, and molecular interactions inside living cells is essential for the cell and molecular biology science. For the purpose of monitoring and investigating molecular events in living cells at real-time, the surface plasmon resonance based cytosensor (SBCS) for cell cultur-ing and signal monitoring was established, and on the basis of it, the corresponding technology was also established by monitoring and analyzing SPR re-sponses induced in rat C6 glioma cells by phorbol 12-myristate 13-acetate (PMA). The SPR signals induced by PMA in living C6 cells were significantly different from those groups without cells. These re-sponses were strongly dependent on and saturable to the concentrations of PMA, and could be sup-pressed by the specific and potent PKC inhibitors, which indicated that the measured signal could be the reflection of the redistribution of intracellular components near the cell membrane triggered by the activation of PKC. This research provides a quantita-tive and non-invasive technique to study the spa-tio-temporal characteristics of the cellular effects in living cells at real-time. Furthermore, this technology could also be widely used in the basic research as well as applied realms, such as space effects evaluation, environmental safety assessment, bio-logical weapon detection, cellular and molecular re-search, and drug screening.
基金financially supported by Major science and technology project of Yunnan Province(202302AE090022)Key Research and Development Program of Yunnan(202203AC100010)+6 种基金the National Natural Science Foundation of China(32160597,32160236,32371463)National Key Research and Development Program of China(2022YFC2601604)Cardiovascular Ultrasound Innovation Team of Yunnan Province(202305AS350021)Spring City Plan:the High-level Talent Promotion and Training Project of Kunming(2022SCP001)the Association Foundation Program of Yunnan Provincial Science and TechnologyDepar tmentKunming Medical University(202101AY070001-278)the second phase of“Double-First Class”program construction of Yunnan University。
文摘Circulating tumor cells(CTCs)are important markers for cancer.The part of tumor cells that are detached from the primary tumor or metastatic tumor and enter the blood circulation is called CTCs.It is crucial to develop a rapid,accurate,and easy-to-implement diagnostic system for CTCs for early tumor diagnosis and for monitoring progression of the disease.In this work,we reported a colorimetric and fluorescent dual-mode assay for the detection of CTCs.Our assay used magnetic nanoparticles and aptamer for CTCs capture and gold nanoparticles-loaded covalent organic frameworks(Au@COFs)for signal amplification,respectively.The magnetic nanoparticles were modified with folic acid to capture CTCs by interaction between folic acid and the folate receptor overexpressed on the surface of tumor cells.The covalent organic frameworks were engineered to have both nitro-reductase-like and glucose–oxidase-like activities.The nitro-reductase-like activity converted the substrate p-nitrophenol to p-nitroaniline for colorimetric detection,and the glucose-oxidase-like activity enabled fluorescence detection.Specifically,Au@COFs catalyzed glucose oxidation and generated hydrogen peroxide to oxidize Fe 2+to Fe 3+,which converted MIL(Al)-MOF to MIL(Fe)-MOF through ion exchange,resulting in the fluorescence quenching of MIL(Al)-MOF.Our assay showed high sensitivity with a detection limit of 17 cells/mL using MCF-7 cells as model cancer cells.This work provided an e fficient and ultrasensitive strategy for CTCs detection and has potential applications in cancer identification and diagnosis.Dual-mode detection system,combining colorimetric and fluorescent signals,integration of magnetic nanoparticles and aptamers for CTC capture,and utilization of covalent organic frameworks loaded with gold nanoparticles for signal amplification can improve sensitivity and accuracy.
