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Published on: May 11, 2016
Isolation and Characterization of Chemo-Resistant Stem Cells from a Mouse Model of Hereditary Non-Polyposis Colon
1Cancer Prevention Research Program, Palindrome Liaisons Consultants, Montvale, NJ 07645-1559, USA.
Rationale:
Loss of function mutations in DNA mismatch repair genes is the primary genetic defects in high-risk hereditary non-polyposis colon cancer (HNPCC). Cytotoxic chemotherapy and anti-inflammatory drugs are potential treatment options. These treatment options lead to systemic toxicity, acquired tumor resistance and the emergence of drug-resistant stem cells. A colonic epithelial cell culture model expressing the relevant genetic defects in chemo-resistant stem cells provides a relevant experimental system for HNPCC.
Objective:
To develop a colonic epithelial cell culture system from a mouse model for HNPCC and to isolate and characterize drug-resistant stem cells.
Experimental Models And Biomarkers:
The Mlh1 [-/-]/Apc [-/-] Mlh1/1638N COL-Cl1 cells is a mouse model for HNPCC, and the 5-fluoro-uracil resistant (5-FU-R) phenotype represents a model for the drug-resistant stem cells. Tumor spheroid formation, and the expression of CD44, CD133 and c-Myc represent stem cell markers.
Results:
The HNPCC model exhibits aneuploidy, hyper-proliferation, accelerated cell cycle progression and downregulated cellular apoptosis. Long-term exposure to 5-FU selects for the drug-resistant phenotype. These resistant cells exhibit increased formation of tumor spheroids and upregulated expression of cancer stem cell markers CD44, CD133 and c-Myc.
Conclusion:
In the present study, a stem cell model for HNPCC was validated and offered a novel experimental approach to test stem cell-targeted alternatives to drug-resistant therapy.
Insights
This study developed a mouse model for hereditary non-polyposis colon cancer (HNPCC) and identified drug-resistant stem cells. These cells show increased tumor spheroid formation and elevated cancer stem cell markers, offering new therapeutic targets.
Area of Science:
- Oncology
- Genetics
- Cell Biology
Background:
- Hereditary non-polyposis colon cancer (HNPCC) is primarily caused by DNA mismatch repair gene mutations.
- Current treatments like chemotherapy face challenges including toxicity, tumor resistance, and drug-resistant stem cells.
- A colonic epithelial cell culture model is crucial for studying HNPCC chemo-resistant stem cells.
Purpose of the Study:
- To establish a colonic epithelial cell culture system using a mouse model for HNPCC.
- To isolate and characterize drug-resistant stem cells within this model.
Main Methods:
- Utilized Mlh1-/-/Apc-/- Mlh1/1638N COL-Cl1 cells as a mouse model for HNPCC.
- Developed a 5-fluoro-uracil resistant (5-FU-R) phenotype to model drug-resistant stem cells.
- Assessed tumor spheroid formation and expression of stem cell markers (CD44, CD133, c-Myc).
Main Results:
- The HNPCC model displayed aneuploidy, hyper-proliferation, and reduced apoptosis.
- Prolonged 5-FU exposure selected for drug-resistant cells.
- Resistant cells demonstrated enhanced tumor spheroid formation and increased CD44, CD133, and c-Myc expression.
Conclusions:
- Validated a stem cell model for HNPCC.
- This model provides a novel platform for investigating stem cell-targeted therapies against drug-resistant HNPCC.
