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Updated: Sep 2, 2025
Simultaneous Imaging and Flow-Cytometry-based Detection of Multiple Fluorescent Senescence Markers in Therapy-Induced Senescent Cancer Cells
Published on: July 12, 2022
Simultaneous Imaging and Flow-cytometry-based Detection of Multiple Fluorescent Senescence Markers in Therapy-induced
Eva Dovjak1, Mario Mairhofer1, Claudia Wöß1
1Johannes Kepler University Linz.
Abstract:
Chemotherapeutic drugs can induce irreparable DNA damage in cancer cells, leading to apoptosis or premature senescence. Unlike apoptotic cell death, senescence is a fundamentally different machinery restraining propagation of cancer cells. Decades of scientific studies have revealed the complex pathological effects of senescent cancer cells in tumors and microenvironments that modulate cancer cells and stromal cells. New evidence suggests that senescence is a potent prognostic factor during cancer treatment, and therefore rapid and accurate detection of senescent cells in cancer samples is essential. This paper presents a method to visualize and detect therapy-induced senescence (TIS) in cancer cells. Diffuse large B-cell lymphoma (DLBCL) cell lines were treated with mafosfamide (MAF) or daunorubicin (DN) and examined for the senescence marker, senescence-associated β-galactosidase (SA-β-gal), the DNA synthesis marker 5-ethynyl-2'-deoxyuridine (EdU), and the DNA damage marker gamma-H2AX (γH2AX). Flow cytometer imaging can help generate high-resolution single-cell images in a short period of time to simultaneously visualize and quantify the three markers in cancer cells.
Insights
Chemotherapy can cause cancer cells to become senescent, a state that stops their growth. This study introduces a new imaging flow cytometry method to quickly detect therapy-induced senescence (TIS) in cancer cells.
Area of Science:
- Oncology
- Cell Biology
- Biotechnology
Background:
- Chemotherapeutic agents induce DNA damage in cancer cells, triggering apoptosis or senescence.
- Senescence, distinct from apoptosis, halts cancer cell proliferation and influences tumor microenvironments.
- Accurate detection of therapy-induced senescence (TIS) is crucial for cancer treatment prognosis.
Purpose of the Study:
- To develop and validate a method for visualizing and quantifying TIS in cancer cells.
- To assess the utility of imaging flow cytometry for simultaneous detection of senescence markers.
Main Methods:
- Diffuse large B-cell lymphoma (DLBCL) cell lines were treated with mafosfamide (MAF) or daunorubicin (DN).
- Cells were analyzed for senescence-associated β-galactosidase (SA-β-gal), 5-ethynyl-2'-deoxyuridine (EdU) for DNA synthesis, and gamma-H2AX (γH2AX) for DNA damage.
- Imaging flow cytometry was employed for high-resolution, single-cell analysis.
Main Results:
- Imaging flow cytometry enabled simultaneous visualization and quantification of SA-β-gal, EdU, and γH2AX in treated DLBCL cells.
- The method demonstrated efficiency in rapidly detecting TIS markers at the single-cell level.
Conclusions:
- Imaging flow cytometry provides a rapid and accurate approach for detecting TIS.
- This technique facilitates the study of senescence as a prognostic factor in cancer therapy.
