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Series GSE24460 Query DataSets for GSE24460
Status Public on Sep 26, 2011
Title Prolonged Drug Selection of Breast Cancer Cells and Enrichment of Cancer Stem Cell Characteristics.
Organism Homo sapiens
Experiment type Expression profiling by array
Summary Background: Cancer stem cells are presumed to have virtually unlimited proliferative and self-renewal abilities and to be highly resistant to chemotherapy, a feature that is associated with overexpression of ATP-binding cassette transporters. We investigated whether prolonged continuous selection of cells for drug resistance enriches cultures for cancer stem-like cells.
Methods: Cancer stem cells were defined as CD44+/CD24– cells that could self-renew (ie, generate cells with the tumorigenic CD44+/CD24– phenotype), differentiate, invade, and form tumors in vivo. We used doxorubicin-selected MCF-7/ADR cells, weakly tumorigenic parental MCF-7 cells, and MCF-7/MDR, an MCF-7 subline with forced expression of ABCB1 protein. Cells were examined for cell surface markers and side-population fractions by microarray and flow cytometry, with in vitro invasion assays, and for ability to form mammospheres. Xenograft tumors were generated in mice to examine tumorigenicity (n = 52). The mRNA expression of multidrug resistance genes was examined in putative cancer stem cells and pathway analysis of statistically significantly differentially expressed genes was performed. All statistical tests were two-sided.
Results: Pathway analysis showed that MCF-7/ADR cells express mRNAs from ABCB1 and other genes also found in breast cancer stem cells (eg, CD44, TGFB1, and SNAI1). MCF-7/ADR cells were highly invasive, formed mammospheres, and were tumorigenic in mice. In contrast to parental MCF-7 cells, more than 30% of MCF-7/ADR cells had a CD44+/CD24– phenotype, could self-renew, and differentiate (ie, produce CD44+/CD24– and CD44+/CD24+ cells), and overexpressed various multidrug resistance-linked genes (including ABCB1, CCNE1, and MMP9). MCF-7/ADR cells were statistically significantly more invasive in Matrigel than parental MCF-7 cells (MCF-7 cells = 0.82 cell per field and MCF-7/ADR = 7.51 cells per field, difference = 6.69 cells per field, 95% confidence interval = 4.82 to 8.55 cells per field, P<.001). No enrichment in the CD44+/CD24– or CD133+ population was detected in MCF-7/MDR.
Conclusion: The cell population with cancer stem cell characteristics increased after prolonged continuous selection for doxorubicin resistance.
Overall design PARALLEL study design with 4 samples Parental MCF-7 cell line versus Doxorubicin Resistant MCF-7 cell sublines Biological replicates: 2 parental controls, 2 drug resistant, independently grown and harvested.
agent:Selection agent is multi-step doxorubicin selection: MCF7226ng, MCF7262ng
biological replicate: MCF71, MCF72
biological replicate: MCF226ng, MCF7262ng
Contributor(s) Calcagno AM, Salcido CD, Guillet J, Wu C, Fostel JM, Mumau MD, Gottesman MM, Varticovski L, Ambudkar SV
Citation(s) 20935265
Submission date Sep 30, 2010
Last update date Dec 06, 2018
Contact name Suresh V. Ambudkar
Organization name National Cancer Institute, NIH
Lab Laboratory of Cell Biology
Street address 37 Convent Dr., Room 2120
City Bethesda
State/province MD
ZIP/Postal code 20892
Country USA
Platforms (1)
GPL571 [HG-U133A_2] Affymetrix Human Genome U133A 2.0 Array
Samples (4)
GSM602658 MCF-7_parental_cell_rep_1
GSM602659 MCF-7_parental_cell_rep_2
GSM602660 MCF-7ADR_multi-step_doxorubicin_selected_subline_rep_1
BioProject PRJNA132705

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GSE24460_RAW.tar 7.5 Mb (http)(custom) TAR (of CEL)
Processed data included within Sample table

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