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Updated: Jun 27, 2025
Assessing Autophagic Flux by Measuring LC3, p62, and LAMP1 Co-localization Using Multispectral Imaging Flow Cytometry
Published on: July 21, 2017
Near-infrared imaging for visualizing the synergistic relationship between autophagy and NFS1 protein during
Wei Hu1,2,3, Yifan He1, Haixian Ren2
1Key Laboratory of Catalysis and Energy Materials Chemistry of Ministry of Education, Key Laboratory of Analytical Chemistry of the State Ethnic Affairs Commission, College of Chemistry and Materials Science, South-Central Minzu University Wuhan 430074 China lichychem@mail.scuec.edu.cn wangyychem@mail.scuec.edu.cn.
Abstract:
Drug resistance is a major challenge for cancer treatment, and its identification is crucial for medical research. However, since drug resistance is a multi-faceted phenomenon, it is important to simultaneously evaluate multiple target fluctuations. Recently developed fluorescence-based probes that can simultaneously respond to multiple targets offer many advantages for real-time and in situ monitoring of cellular metabolism, including ease of operation, rapid reporting, and their non-invasive nature. As such we developed a dual-response platform (Vis-H2S) with integrated ICT-TICT to image H2S and viscosity in mitochondria, which could simultaneously track fluctuations in cysteine desulfurase (NFS1 protein and H2S inducer) and autophagy during chemotherapy-induced multidrug resistance. This platform could monitor multiple endogenous metabolites and the synergistic relationship between autophagy and NFS1 protein during multidrug resistance induced by chemotherapy. The results indicated that chemotherapeutic drugs simultaneously up-regulate the levels of NFS1 protein and autophagy. It was also found that the NFS1 protein was linked with autophagy, which eventually led to multidrug resistance. As such, this platform could serve as an effective tool for the in-depth exploration of drug resistance mechanisms.
Insights
This study introduces a new dual-response platform to image hydrogen sulfide (H₂S) and viscosity in mitochondria. It tracks key factors in chemotherapy-induced drug resistance, revealing a link between NFS1 protein and autophagy.
Area of Science:
- Biochemistry
- Cell Biology
- Cancer Research
Background:
- Drug resistance is a significant hurdle in cancer therapy, necessitating methods to monitor complex cellular changes.
- Simultaneous evaluation of multiple cellular targets is crucial for understanding multifaceted drug resistance.
- Fluorescence-based probes offer real-time, in situ monitoring of cellular metabolism.
Purpose of the Study:
- To develop a dual-response platform for simultaneous imaging of hydrogen sulfide (H₂S) and viscosity in mitochondria.
- To investigate the synergistic relationship between autophagy and NFS1 protein in chemotherapy-induced multidrug resistance.
- To track fluctuations in cysteine desulfurase (NFS1) and autophagy during cancer drug resistance.
Main Methods:
- Development of a dual-response platform (Vis-H₂S) integrating ICT-TICT.
- In situ imaging of H₂S and viscosity within mitochondria.
- Simultaneous monitoring of NFS1 protein and autophagy levels.
Main Results:
- Chemotherapeutic drugs were found to up-regulate both NFS1 protein and autophagy levels.
- A direct link between NFS1 protein and autophagy was identified as a contributor to multidrug resistance.
- The platform successfully monitored endogenous metabolites and their synergistic interactions.
Conclusions:
- The developed Vis-H₂S platform is effective for real-time, in situ monitoring of cellular metabolism and drug resistance mechanisms.
- The study elucidates the interconnected roles of NFS1 protein and autophagy in the development of multidrug resistance.
- This tool facilitates deeper exploration into the complexities of cancer drug resistance.
