Abstract
Cancer stem-like cells (CSCs) are a typically small subpopulation of highly tumorigenic cells that can self-renew, differentiate, drive tumor progression, and may mediate drug resistance and metastasis. Metastasis driving CSCs are expected to be highly invasive. To determine the relative invasiveness of CSCs, we isolate distinct subpopulations in the metastatic, MDA-MB-231 breast-cancer cell line, identified by the stem-cell markers aldehyde dehydrogenase (ALDH) and CD44. We determine CSC-subpopulation invasiveness levels using our rapid (2 h) mechanobiology-based assay. Specifically, invasive cells forcefully push and indent the surface of physiological–stiffness synthetic gels to cell-scale depths, where the percentage of indenting cells and their attained depths have previously provided clinically relevant predictions of the metastatic risk in different cancer types. We observe that the small (3.2%) CD44+ALDH+ cell-subpopulation indents more and attains significantly deeper depths (65% indenting to 6 ± 0.3 µm) relative to CD44+ALDH−, CD44−ALDH−, CD44−ALDH+ cells, and the whole-sample control (with 18–44% indenting cells reaching average depths of 4.4–5 µm). The CD44+ALDH+ similarly demonstrates twofold higher migratory capacity in Boyden chambers. The higher invasiveness of CD44+ALDH+ cells reveals their likely role in facilitating disease progression, providing prognostic markers for increased risk of recurrence and metastasis.
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Data availability
The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.
Abbreviations
- CSCs:
-
Cancer stem-like cells
- ALDH:
-
Aldehyde dehydrogenase
- DMEM:
-
Dulbecco’s modified Eagle’s medium
- FBS:
-
Fetal bovine serum
- APS:
-
Ammonium persulfate
- TEMED:
-
Tetramethylethylenediamine
- ANOVA:
-
Analysis of variance
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Acknowledgements
The work was partially supported by the Israeli Ministry of Science and Technology (MOST) Medical Devices Program (Grant no. 3-17427), by the Samuel H. Born Fund for Biomedical Research, and by the Russel Berrie Nano Institute at the Technion-IIT.
Funding
The work was partially supported by the Israeli Ministry of Science and Technology (MOST) Medical Devices Program (Grant no. 3–17427), the Samuel H. Born Fund for Biomedical Research, and by the Russel Berrie Nano Institute at the Technion-IIT.
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MBA-E and DW contributed to the study conception and design, material preparation, data collection and analysis. The first draft of the manuscript was written by MBA-E and DW.
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Alvarez-Elizondo, M.B., Weihs, D. Breast cancer stem cells: mechanobiology reveals highly invasive cancer cell subpopulations. Cell. Mol. Life Sci. 79, 134 (2022). https://doi.org/10.1007/s00018-022-04181-w
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DOI: https://doi.org/10.1007/s00018-022-04181-w