Mitochondria/lysosome dual-organelle labelling esterase probe for monitoring cell viability and evaluating lung cancer drug efficiency

Spectrochim Acta A Mol Biomol Spectrosc. 2025 Feb 15:327:125379. doi: 10.1016/j.saa.2024.125379. Epub 2024 Nov 2.

Abstract

Monitoring of cell viability plays a key role in cancer therapy and evaluation of drug efficiency. Mitochondria and lysosomes are involved in regulating cell viability in many biological processes such as apoptosis, necrosis, autophagy, and cell proliferation. Thus, there is an emerging interest in the real-time evaluation of cell viability in both mitochondria and lysosomes. Herein, for the first time, we rationally designed and developed a mitochondria/lysosome dual-organelle labelling esterase-responsive ratiometric fluorescent probe, named TMLE-2, for dual-channel monitoring of cell viability and evaluation of lung cancer drug efficiency. TMLE-2 showed dramatic ratio fluorescence changes (about 51-fold) upon reacting with esterase. Furthermore, TMLE-2 enabled visualization of mitochondria and lysosomes with red and green emission, respectively; moreover, H2O2-induced cell damage, sorafenib-induced ferroptosis and ascorbic-acid-mediated cell protective effects were successfully assessed by dual-organelle ratiometric fluorescent imaging and flow cytometry data. More importantly, TMLE-2 was successfully used for the first time to evaluate the efficiency of lung cancer drugs at the cellular and tissue levels based on dual-organelle esterase activity assay. In summary, the newly designed TMLE-2 is expected to have enormous potential for facilitating advancements in biomedical fields related to cell viability.

Keywords: Cell viability; Dual organelles imaging; Esterase probe; Ratiometric fluorescent imaging; Single fluorescent probe.

MeSH terms

  • A549 Cells
  • Antineoplastic Agents* / pharmacology
  • Cell Line, Tumor
  • Cell Survival* / drug effects
  • Esterases* / metabolism
  • Ferroptosis / drug effects
  • Fluorescent Dyes* / chemistry
  • Humans
  • Hydrogen Peroxide / metabolism
  • Lung Neoplasms* / drug therapy
  • Lung Neoplasms* / pathology
  • Lysosomes* / drug effects
  • Lysosomes* / metabolism
  • Mitochondria* / drug effects
  • Mitochondria* / metabolism

Substances

  • Esterases
  • Fluorescent Dyes
  • Antineoplastic Agents
  • Hydrogen Peroxide