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Published on: August 16, 2021
Dynamic Monitoring of Content Variation of Guanosine Triphosphate in Single Cells under External Stimulus with
Xin Guan1, Wei Zhao1, Wanjiao Li1
1School of Basic Medical Sciences, Beihua University, Jilin, 132013 Jilin, P. R. China.
Abstract:
Photothermal therapy (PTT) has demonstrated significant potential in the ablation of tumors, but understanding of the dynamic molecular changes in intracellular energy metabolism during PTT-induced cell death is less studied, which is helpful for the optimization of the therapeutic approach. Guanosine triphosphate (GTP), is a key nucleotide involved in energy metabolism and cellular processes, playing a critical role in tumor cell proliferation and metabolic reprogramming. Herein, we developed an aptamer-functionalized Nb2C MXene nanoprobe (FA-Nb2C-GTP) to monitor dynamic content variation of intracellular GTP during PTT process. Based on fluorescence resonance energy transfer (FRET), the probe exhibits an ″off-on″ fluorescence response upon GTP binding. It exhibits high specificity for GTP (LOD = 15.67 μM), good photothermal conversion efficiency (34.21%), and biocompatibility. During photothermal therapy, FA-Nb2C-GTP enabled real-time cell imaging of dynamic content variation of GTP. In HepG2 cells, the experiments found that GTP levels initially increased, reaching a peak at 5 min postirradiation with a 232.8% fluorescence enhancement relative to controls, followed by a decline upon extended NIR irradiation exposure. Conversely, HL-7702 cells exhibited minimal GTP fluctuations, with a maximum fluorescence increase of 23.8% at 7 min postirradiation. These results revealed distinct GTP metabolic response behaviors between the tumor and normal cells during PTT-induced apoptosis. The designed FA-Nb2C-GTP nanoprobe represents a powerful tool for investigating the relationship between energy metabolism and PTT-induced tumor cell death, offering insights for the rational design of targeted PTT agents.
Insights
Researchers developed a novel aptamer-functionalized MXene nanoprobe to monitor guanosine triphosphate (GTP) levels during photothermal therapy (PTT). This tool reveals distinct intracellular GTP metabolic responses in tumor versus normal cells, aiding PTT optimization.
Area of Science:
- Biomedical Engineering
- Nanotechnology
- Molecular Biology
Background:
- Photothermal therapy (PTT) shows promise for tumor ablation, but intracellular metabolic changes during PTT-induced cell death require further study.
- Guanosine triphosphate (GTP) is crucial for energy metabolism and tumor cell proliferation, making its dynamic changes during PTT a key area of investigation.
- Understanding metabolic reprogramming during PTT is essential for optimizing therapeutic strategies and developing targeted agents.
Purpose of the Study:
- To develop a nanoprobe for real-time monitoring of intracellular guanosine triphosphate (GTP) variations during photothermal therapy (PTT).
- To investigate the distinct metabolic responses of tumor and normal cells to PTT by tracking dynamic GTP content.
- To establish a tool for understanding the relationship between energy metabolism and PTT-induced cell death.
Main Methods:
- Development of an aptamer-functionalized Nb2C MXene nanoprobe (FA-Nb2C-GTP) utilizing fluorescence resonance energy transfer (FRET) for GTP detection.
- Characterization of the probe's specificity, limit of detection (LOD = 15.67 μM), photothermal conversion efficiency (34.21%), and biocompatibility.
- Real-time cell imaging to monitor intracellular GTP dynamics in HepG2 (tumor) and HL-7702 (normal) cells during PTT.
Main Results:
- The FA-Nb2C-GTP probe exhibited an "off-on" fluorescence response specific to GTP.
- HepG2 cells showed a significant initial increase in GTP levels post-irradiation, peaking at 5 min (232.8% fluorescence enhancement), followed by a decline.
- HL-7702 cells displayed minimal GTP fluctuations, with a maximum fluorescence increase of 23.8% at 7 min, indicating distinct metabolic responses between tumor and normal cells.
Conclusions:
- The FA-Nb2C-GTP nanoprobe effectively enables real-time monitoring of intracellular GTP dynamics during PTT.
- Distinct GTP metabolic responses observed in tumor versus normal cells provide insights into PTT-induced apoptosis mechanisms.
- This nanoprobe serves as a valuable tool for investigating energy metabolism in PTT and designing improved therapeutic agents.
