Expression profiling of stem cell signaling alters with spheroid formation in CD133high/CD44high prostate cancer stem cells
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Date
2014
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Abstract
Cancer stem cells (CSC) isolated from multiple tumor types differentiate in vivo and in vitro when cultured in serum; however, the factors responsible for their differentiation have not yet been identified. The first aim of the present study was to identify CD133high/CD44high DU145 prostate CSCs and compare their profiles with non-CSCs as bulk counterparts of the population. Subsequently, the two populations continued to be three-dimensional multicellular spheroids. Differentiation was then investigated with stem cell-related genomic characteristics. Polymerase chain reaction array analyses of cell cycle regulation, embryonic and mesenchymal cell lineage-related markers, and telomerase reverse transcriptase (TERT) and Notch signaling were performed. Immunohistochemistry of CD117, Notch1, Jagged1, Delta1, Sox2, c-Myc, Oct4, KLF4, CD90 and SSEA1 were determined in CSC and non-CSC monolayer and spheroid subcultures. Significant gene alterations were observed in the CD133high/CD44high population when cultured as a monolayer and continued as spheroid. In this group, marked gene upregulation was determined in collagen type 9 α1, Islet1 and cyclin D2. Jagged1, Delta-like 3 and Notch1 were respectively upregulated genes in the Notch signaling pathway. According to immunoreactivity, the staining density of Jagged1, Sox2, Oct4 and Klf-4 increased significantly in CSC spheroids. Isolated CSCs alter their cellular characterization over the course of time and exhibit a differentiation profile while maintaining their former surface antigens at a level of transcription or translation. The current study suggested that this differentiation process may be a mechanism responsible for the malignant process and tumor growth.
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Keywords
CD133 antigen , collagen type 9 , complementary DNA , cyclin D2 , Delta1 , Hermes antigen , Jagged1 , kruppel like factor 4 , Myc protein , Notch1 receptor , octamer transcription factor 4 , stage specific embryo antigen 1 , stem cell factor receptor , telomerase reverse transcriptase , Thy 1 antigen , transcription factor , transcription factor Sox2 , unclassified drug , article , cancer cell culture , cancer stem cell , cell cycle regulation , cell differentiation , cell function , cell lineage , controlled study , fluorescence activated cell sorting , functional genomics , gene expression , human , human cell , immunohistochemistry , microarray analysis , polymerase chain reaction , prostate cancer , RNA isolation , tumor growth , tumor spheroid