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Updated: Jun 17, 2026
Isolation and Functional Assessment of Human Breast Cancer Stem Cells from Cell and Tissue Samples
Published on: October 2, 2020
Regulation of breast cancer stem cell activity by signaling through the Notch4 receptor
Hannah Harrison1, Gillian Farnie, Sacha J Howell
1Breast Biology Group, School of Cancer, Enabling Sciences and Technology, Paterson Institute for Cancer Research, University of Manchester, Manchester Academic Health Sciences Centre, The Christie NHS Foundation Trust; Department of Medical Oncology, The Christie NHS Foundation Trust, Manchester M20 4BX, United Kingdom.
Abstract:
Notch receptor signaling pathways play an important role not only in normal breast development but also in breast cancer development and progression. We assessed the role of Notch receptors in stem cell activity in breast cancer cell lines and nine primary human tumor samples. Stem cells were enriched by selection of anoikis-resistant cells or cells expressing the membrane phenotype ESA(+)/CD44(+)/CD24(low). Using these breast cancer stem cell populations, we compared the activation status of Notch receptors with the status in luminally differentiated cells, and we evaluated the consequences of pathway inhibition in vitro and in vivo. We found that Notch4 signaling activity was 8-fold higher in stem cell-enriched cell populations compared with differentiated cells, whereas Notch1 signaling activity was 4-fold lower in the stem cell-enriched cell populations. Pharmacologic or genetic inhibition of Notch1 or Notch4 reduced stem cell activity in vitro and reduced tumor formation in vivo, but Notch4 inhibition produced a more robust effect with a complete inhibition of tumor initiation observed. Our findings suggest that Notch4-targeted therapies will be more effective than targeting Notch1 in suppressing breast cancer recurrence, as it is initiated by breast cancer stem cells.
Insights
Notch4 signaling is elevated in breast cancer stem cells, driving tumor growth. Inhibiting Notch4 effectively suppressed tumor initiation, suggesting it as a promising therapeutic target for preventing breast cancer recurrence.
Area of Science:
- Oncology
- Molecular Biology
- Cell Biology
Background:
- Notch receptor signaling is crucial in normal breast development and cancer.
- Breast cancer stem cells (BCSCs) drive tumor progression and recurrence.
- Understanding Notch pathway roles in BCSCs is vital for targeted therapies.
Purpose of the Study:
- To investigate the role of Notch receptors (Notch1 and Notch4) in breast cancer stem cell activity.
- To compare Notch signaling activation in BCSCs versus differentiated breast cancer cells.
- To evaluate the therapeutic potential of inhibiting Notch1 and Notch4 in breast cancer.
Main Methods:
- Enrichment of BCSCs using anoikis-resistance or ESA(+)/CD44(+)/CD24(low) phenotype.
- Comparison of Notch receptor activation status in BCSCs and differentiated cells.
- In vitro and in vivo evaluation of Notch1 and Notch4 inhibition effects on stem cell activity and tumor formation.
Main Results:
- Notch4 signaling was 8-fold higher, while Notch1 signaling was 4-fold lower in BCSCs compared to differentiated cells.
- Pharmacologic or genetic inhibition of Notch1 or Notch4 reduced BCSCs activity and tumor formation.
- Notch4 inhibition demonstrated a more potent effect, completely inhibiting tumor initiation.
Conclusions:
- Notch4 signaling is significantly upregulated in breast cancer stem cells and drives tumor initiation.
- Targeting Notch4 offers a more effective strategy than targeting Notch1 for suppressing breast cancer recurrence.
- Notch4-targeted therapies hold promise for treating breast cancer by eliminating cancer stem cells.
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