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TISSUE-SPECIFIC STEM CELLS

Transcriptional Profiling of Mammary Gland Side Population Cells

^a Department of Molecular and Cellular Biology,
^b Department of Molecular and Human Genetics, and
^c Breast Center, Baylor College of Medicine, Houston, Texas, USA

Key Words. Mammary gland side population cells • Murine and human • Microarray gene profiling

Correspondence: Jeffrey Rosen, Ph.D., C.C. Bell Professor of Molecular and Cellular Biology and Medicine, DeBakey Building, M638a, Baylor College of Medicine, One Baylor Plaza, Houston, Texas 77030-3498, USA. Telephone: 713-798-6210; Fax 713-798-8012; e-mail: jrosen{at}bcm.tmc.edu

Received August 5, 2005; accepted for publication November 4, 2005.
Similar to the bone marrow, the mammary gland contains a distinct population of Hoechst-effluxing side population cells, mammary gland side population cells (MG-SPs). To better characterize MG-SPs, their microarray gene profiles were compared to the remaining cells, which retain Hoechst dye (mammary gland non-side population cells [MG-NSPs]). For analysis, Gene Ontology (GO) that describes genes in terms of biological processes and Ontology Traverser (OT) that performs enrichment analysis were used. OT showed that MG-SP-specific genes were enriched in the GO categories of cell cycle regulation and checkpoints, multidrug-resistant transporters, organogenesis, and vasculogenesis. The MG-NSP-upregulated genes were enriched in the GO category of cellular organization and biogenesis, which includes basal epithelial markers, p63, smooth muscle actin, myosin, 6 integrin, cytokeratin (CK) 14, and luminal markers CK8 and CD24. Additional studies showed that a higher percentage of MG-SPs exist in the G1 phase of the cell cycle compared with the MG-NSPs. G1 cell cycle block of MG-SPs may be explained by higher expression of cell cycle-negative regulatory genes such as transforming growth factor-ß2, insulin-like growth factor binding protein-5, P18^INK4C, and wingless-5a (Wnt-5a). Accordingly, a smaller percentage of MG-SPs expressed nuclear ß-catenin, possibly as a consequence of the higher expression of Wnt-5a. In conclusion, microarray gene profiling suggests that MG-SPs are a lineage-deficient mammary gland subpopulation expressing key genes involved in cell cycle regulation, development, and angiogenesis.

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