Journal: Cancers
Article Title: The Cationic Amphiphilic Drug Hexamethylene Amiloride Eradicates Bulk Breast Cancer Cells and Therapy-Resistant Subpopulations with Similar Efficiencies
doi: 10.3390/cancers14040949
Figure Lengend Snippet: HMA ablates chemotherapy-resistant BCSCs but not normal mammary stem-like cells. ( A ) Secondary tumorsphere formation by human breast cancer cell lines was assessed after pretreatment with various concentrations of HMA for 24 h. Data are representative of three independent experiments. ( B ) Secondary sphere formation by CD44 + /CD24 low BCSC-enriched human breast cancer cells after HMA pretreatment. ( C ) Secondary mammosphere formation of non-transformed human and mouse mammary epithelial cell lines was assessed after 40 μM HMA treatment for 24 h. ( D ) Total populations of MDA-MB-231, MCF7, SKBR3, and T47D cells were treated with vehicle (DMSO), 40 μM HMA, 40 μM cisplatin (CIS), or a combination of 170 nM doxorubicin (DOX) and 50 nM docetaxel (DTX) for 24 h, and cell viability was determined by trypan blue exclusion assay. ( E ) Sorted human BCSC cells were treated with vehicle, 40 μM HMA, 40 μM CIS, or 170 nM DOX with 50 nM DTX, and secondary sphere formation was determined. ( F , G ) Cells dissociated from MMTV-NDL mouse mammary tumors were sorted (CD24 high /CD49f high /Lin − ) to enrich for the BCSC population, and 7-day sphere formation following pretreatment with HMA ( F ) or standard chemotherapeutics ( G ) was quantified over four biological tumor replicates from independent mice. Error bars represent SEM. *, p < 0.05; **, p < 0.01; ***, p < 0.001, by Student’s t -test.
Article Snippet: HMEC4 cells were maintained in mammary epithelial basal media (MEBM; #CC-3151, Lonza, Basel, Switzerland) with MEGM SingleQuots Supplements (#CC-4136, Lonza).
Techniques: Transformation Assay, Trypan Blue Exclusion Assay
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