Related Experiment Video
Updated: Jun 8, 2026
04:47
Autonomously Bioluminescent Mammalian Cells for Continuous and Real-time Monitoring of Cytotoxicity
Published on: October 28, 2013
A Color-Tuning Bioluminescent Sensor (AmyLuc) for Real-Time Monitoring of Intracellular pH Dynamics in Cancer Cells
Vanessa R Bevilaqua1,2,3, Angela Punzo2,4, Alessia Silla2,4
1Laboratory of Biomaterials, Department of Surgery, Pontifical Catholic University of São Paulo (PUC-SP), Sorocaba, São Paulo 18030-070, Brazil.
Analytical Chemistry
|June 7, 2026
Summary
Cancer cells reprogram metabolism, causing pH changes crucial for tumor growth. A new firefly luciferase biosensor (AmyLuc) effectively monitors these intracellular and extracellular pH shifts in real-time.
Area of Science:
- Biochemistry
- Molecular Biology
- Cancer Research
Background:
- Cancer cells exhibit altered metabolism, characterized by increased glucose uptake and lactate production (Warburg effect).
- This metabolic shift leads to tumor microenvironment (TME) acidification and intracellular alkalinization, promoting malignant behaviors like invasion and immune evasion.
- Real-time monitoring of intra- and extracellular pH dynamics is critical for understanding tumor progression and assessing therapeutic efficacy.
Purpose of the Study:
- To develop and validate a novel pH-sensitive bioluminescent biosensor for real-time monitoring of intracellular and extracellular pH changes in cancer cells.
- To investigate the metabolic alterations and associated pH dynamics in colorectal adenocarcinoma cells (Caco-2) under specific treatments.
Main Methods:
- Utilized a pH-sensitive bioluminescent color-tuning biosensor derived from firefly *Amydetes vivianii* luciferase (AmyLuc).
- Established a calibration curve using the ratio of bioluminescence emission intensities at 593 nm (pH 6.0) and 548 nm (pH 8.0) for accurate pH determination.
- Applied treatments with carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP) and antimycin A to induce metabolic stress in Caco-2 cells.
Main Results:
- AmyLuc biosensor successfully ratiometrically estimated intracellular and extracellular pH changes.
- Treatments induced sustained intracellular acidification (pH ~6.3) and gradual extracellular alkalinization (pH ~7.1).
- Observed pH dynamics are consistent with metabolic reprogramming from mitochondrial respiration to anaerobic glycolysis.
Conclusions:
- AmyLuc serves as a sensitive color-tuning bioluminescent pH biosensor for real-time monitoring of pH dynamics in cancer cells.
- The biosensor is suitable for studying cancer cell metabolism under stress and evaluating therapeutic interventions.
- The findings underscore the importance of pH monitoring in cancer research and treatment strategies.
