What is it about?
Studying how cancer cells die requires reliable ways to both induce and measure cell death. In this laboratory protocol, we developed methods for inducing programmed cell death in solid tumour cell lines using chemical treatment and acidic conditions that mimic aspects of the tumour microenvironment. We also optimised Annexin V/propidium iodide flow cytometry to distinguish viable cells from those undergoing early or late programmed cell death and from dead or necrotic cells.
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Photo by National Cancer Institute on Unsplash
Why is it important?
The tumour microenvironment is very different from the conditions typically used to grow cancer cells in the laboratory. Tumours are often acidic, and this environmental stress can influence how cancer cells survive and die. A reproducible method for exposing cells to low pH provides a useful experimental model for investigating these effects. Combining this approach with Annexin V/PI flow cytometry allows different stages of cell death to be quantified at the level of individual cells and cell populations.
Perspectives
Understanding how cancer cells respond to environmental stress is important for studying tumour biology and treatment response. The methods described here provide a relatively accessible approach for investigating programmed cell death under acidic conditions and can be adapted alongside other flow-cytometric markers to examine different aspects of cell behaviour. More broadly, incorporating features of the tumour microenvironment into experimental models may help bridge the gap between conventional cell culture conditions and the complex environment experienced by tumour cells in vivo.
Dr Catherine Worsley
University of Pretoria
Read the Original
This page is a summary of: Inducing apoptosis using chemical treatment and acidic pH, and detecting it using the Annexin V flow cytometric assay, PLOS One, June 2022, PLOS,
DOI: 10.1371/journal.pone.0270599.
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Resources
The effect of acute acid exposure on immunomodulatory protein secretion, cell survival, and cell cycle progression in tumour cell lines
The acidic tumour microenvironment influences tumour cell survival, proliferation, and immune evasion. This article presents data on the effect of low pH on breast and oesophageal squamous cell carcinoma cell (OSCC) lines. Distinct differences in apoptosis were noted between the cell lines, with OSCC better adapted to survive acidic conditions and release pro-tumourigenic compounds. This highlights the important role that tumour acidity plays in driving the evolution of different tumour types.
The acidic tumour microenvironment: Manipulating the immune response to elicit escape
This review article highlights the role that the acidic tumour microenvironment plays in driving immune escape. It describes the effects on different immune cell types, and also discusses therapeutic options to target the acidic tumour microenvironment.
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