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Updated: Dec 10, 2025
Immunofluorescence Imaging of DNA Damage and Repair Foci in Human Colon Cancer Cells
Published on: June 9, 2020
Single-Cell Transcriptome Analysis of Colon Cancer Cell Response to 5-Fluorouracil-Induced DNA Damage
Sung Rye Park1, Sim Namkoong2, Leon Friesen3
1Department of Molecular & Integrative Physiology and Institute for Gerontology, University of Michigan Medical School, Ann Arbor, MI 48109, USA; Department of Biostatistics and Center for Statistical Genetics, University of Michigan School of Public Health, Ann Arbor, MI 48109, USA.
Abstract:
DNA damage often induces heterogeneous cell-fate responses, such as cell-cycle arrest and apoptosis. Through single-cell RNA sequencing (scRNA-seq), we characterize the transcriptome response of cultured colon cancer cell lines to 5-fluorouracil (5FU)-induced DNA damage. After 5FU treatment, a single population of colon cancer cells adopts three distinct transcriptome phenotypes, which correspond to diversified cell-fate responses: apoptosis, cell-cycle checkpoint, and stress resistance. Although some genes are regulated uniformly across all groups of cells, many genes showed group-specific expression patterns mediating DNA damage responses specific to the corresponding cell fate. Some of these observations are reproduced at the protein level by flow cytometry and are replicated in cells treated with other 5FU-unrelated genotoxic drugs, camptothecin and etoposide. This work provides a resource for understanding heterogeneous DNA damage responses involving fractional killing and chemoresistance, which are among the major challenges in current cancer chemotherapy.
Insights
DNA damage triggers diverse colon cancer cell fates, including apoptosis, cell-cycle arrest, and stress resistance. This study reveals distinct gene expression patterns underlying these varied responses to chemotherapy.
Area of Science:
- Molecular Biology
- Cancer Research
- Genomics
Background:
- DNA damage is a known inducer of varied cellular responses, including apoptosis and cell-cycle arrest.
- Understanding the heterogeneity in these responses is crucial for improving cancer chemotherapy efficacy.
- Colon cancer cells exhibit complex reactions to genotoxic stress, impacting treatment outcomes.
Purpose of the Study:
- To characterize the transcriptome response of colon cancer cell lines to 5-fluorouracil (5FU)-induced DNA damage at a single-cell level.
- To identify distinct cell phenotypes and their associated gene expression patterns following DNA damage.
- To explore the implications of heterogeneous cell-fate responses for cancer chemoresistance.
Main Methods:
- Single-cell RNA sequencing (scRNA-seq) was employed to analyze gene expression profiles.
- Colon cancer cell lines were treated with 5-fluorouracil (5FU) to induce DNA damage.
- Flow cytometry was used to validate protein-level changes, and responses to other genotoxic drugs (camptothecin, etoposide) were assessed.
Main Results:
- A single population of colon cancer cells differentiated into three distinct transcriptome phenotypes post-5FU treatment.
- These phenotypes corresponded to specific cell-fate outcomes: apoptosis, cell-cycle checkpoint, and stress resistance.
- Group-specific gene expression patterns were identified, mediating DNA damage responses unique to each cell fate, with some findings validated at the protein level and with other drugs.
Conclusions:
- Colon cancer cells exhibit heterogeneous responses to DNA damage, leading to diversified cell fates.
- Distinct transcriptional programs underlie these varied responses, contributing to fractional killing and chemoresistance.
- This research provides a valuable resource for understanding and potentially overcoming challenges in cancer chemotherapy.
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