Antitumor activity of cell-penetrant kinin B1 receptor antagonists in human triple-negative breast cancer cells.

Abstract

High nuclear expression of G protein-coupled receptors, including kinin B1 receptors (B1R), has been observed in several human cancers, but the clinical significance of this is unknown. We put forward the hypothesis that these "nuclearized" kinin B1R contribute to tumorigenicity and can be a new target in anticancer strategies. Our initial immunostaining and ultrastructural electron microscopy analyses demonstrated high B1R expression predominantly located at internal/nuclear compartments in the MDA-MB-231 triple-negative breast cancer (TNBC) cell line as well as in clinical samples of patients with TNBC. On the basis of these findings, in the present study, we evaluated the anticancer therapeutic potential of newly identified, cell-permeable B1R antagonists in MDA-MB-231 cells (ligand-receptor binding/activity assays and LC-MS/MS analyses). We found that these compounds (SSR240612, NG67, and N2000) were more toxic to MDA-MB-231 cells in comparison with low- or non-B1R expressing MCF-10A normal human mammary epithelial cells and COS-1 cells, respectively (clonogenic, MTT proliferative/cytocidal assays, and fluorescence-activated cell-sorting (FACS)-based apoptosis analyses). By comparison, the peptide B1R antagonist R954 unable to cross cell membrane failed to produce anticancer effects. Furthermore, the putative mechanisms underlying the anticancer activities of cell-penetrant B1R antagonists were assessed by analyzing cell cycle regulation and signaling molecules related to cell survival and apoptosis (FACS and western blot). Finally, drug combination experiments showed that cell-penetrant B1R antagonists can cooperate with suboptimal doses of chemotherapeutic agents (doxorubicin and paclitaxel) to promote TNBC death. This study provides evidence on the potential value of internally acting kinin B1R antagonists in averting growth of breast cancer.

• Authors: Céléna Dubuc, Martin Savard, Veronica Bovenzi, Andrée Lessard, Jérôme Côté, Witold Neugebauer, Sameh Geha, Sylvain Chemtob, Fernand Gobeil Jr
• Journal: Journal of cellular physiology (J Cell Physiol) Vol. 234 Issue 3 Pg. 2851-2865 (03 2019) ISSN: 1097-4652 [Electronic] United States
• PMID: 30132865 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2018 Wiley Periodicals, Inc.

Chemical References

• Bradykinin B1 Receptor Antagonists
• Cell-Penetrating Peptides
• Receptor, Bradykinin B1
• Receptors, G-Protein-Coupled
• Doxorubicin
• Paclitaxel

Topics

• Animals
• Antineoplastic Combined Chemotherapy Protocols (pharmacology)
• Apoptosis (drug effects)
• Bradykinin B1 Receptor Antagonists (pharmacology)
• COS Cells
• Cell Line, Tumor
• Cell Membrane Permeability (drug effects, genetics)
• Cell-Penetrating Peptides (pharmacology)
• Chlorocebus aethiops
• Doxorubicin (pharmacology)
• Female
• Gene Expression Regulation, Neoplastic (drug effects)
• Humans
• Paclitaxel (pharmacology)
• Receptor, Bradykinin B1 (genetics)
• Receptors, G-Protein-Coupled (genetics)
• Signal Transduction (drug effects)
• Triple Negative Breast Neoplasms (drug therapy, genetics, pathology)