HPI/AMF inhibition halts the development of the aggressive phenotype of breast cancer stem cells.

Abstract

Cancer stem cells are responsible for tumor recurrence and metastasis. A new highly reproducible procedure for human breast cancer MCF-7 stem cells (BCSC) isolation and selection was developed by using a combination of hypoxia/hypoglycemia plus taxol and adriamycin for 24h. The BCSC enriched fraction (i) expressed (2-15 times) the typical stemness protein markers CD44+, ALDH1A3 and Oct 3/4; (ii) increased its clonogenicity index (20-times), invasiveness profile (>70%), migration capacity (100%) and ability to form mammospheres, compared to its non-metastatic MCF-7 counterpart. This isolation and selection protocol was successful to obtain stem cell enriched fractions from A549, SiHa and medulloblastoma cells. Since the secretion of HPI/AMF cytokine seems involved in metastasis, the effects of erytrose-4-phosphate (E4P) and 6-phosphogluconate (6PG), potent HPI inhibitors, on the acquisition of the breast stem cell-like phenotype were also evaluated. The presence of E4P during the BCSC selection deterred the development of the stemness phenotype, whereas both extracellular E4P (5-250nM) and 6PG (1μM) as well as siRNA HPI/AMF depressed the BCSC invasiveness ability (>90%), clonogenicity index (>90%) and contents (50-96%) of stemness (CD44, ALDH1A), pluripotency (p38 MAPK, Oct3/4, wnt/β-catenin) and EMT (SNAIL, MMP-1, vimentin) markers. The cytokine inhibitor repertaxin (10nM) or the anti-IL-8 or anti-TGF-β monoclonal antibodies (10μg/mL) did not significantly affect the BCSC metastatic phenotype. E4P also diminished (75%) the formation and growth of MCF-7 stem cell mammospheres. These results suggested that E4P by directly interacting with extracellular HPI/AMF may be an effective strategy to deter BCSC growth and progression.

• Authors: Juan Carlos Gallardo-Pérez, Alhelí Adán-Ladrón de Guevara, Alvaro Marín-Hernández, Rafael Moreno-Sánchez, Sara Rodríguez-Enríquez
• Journal: Biochimica et biophysica acta. Molecular cell research (Biochim Biophys Acta Mol Cell Res) Vol. 1864 Issue 10 Pg. 1679-1690 (Oct 2017) ISSN: 0167-4889 [Print] Netherlands
• PMID: 28648642 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2017 Elsevier B.V. All rights reserved.

Chemical References

• GPS2 protein, human
• Gluconates
• Intracellular Signaling Peptides and Proteins
• RNA, Small Interfering
• Sugar Phosphates
• Sulfonamides
• Doxorubicin
• erythrulose 4-phosphate
• Paclitaxel
• reparixin
• 6-phosphogluconic acid

Topics

• Breast Neoplasms (drug therapy, genetics, pathology)
• Cell Hypoxia (drug effects)
• Cell Movement (drug effects)
• Cell Proliferation (drug effects, genetics)
• Doxorubicin (administration & dosage)
• Epithelial-Mesenchymal Transition (drug effects, genetics)
• Female
• Gluconates (administration & dosage)
• Humans
• Intracellular Signaling Peptides and Proteins (antagonists & inhibitors, genetics)
• MCF-7 Cells
• Neoplasm Metastasis
• Neoplastic Stem Cells (drug effects, pathology)
• Paclitaxel (administration & dosage)
• RNA, Small Interfering (genetics)
• Sugar Phosphates (administration & dosage)
• Sulfonamides (administration & dosage)