Expression profiling of stem cell signaling alters with spheroid formation in CD133high/CD44high prostate cancer stem cells

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.

Notch signaling in the prostate: critical roles during development and in the hallmarks of prostate cancer biology

“…In gene profiling of DU145 prostate cancer cells, a subpopulation with CD133 high /CD44 high was isolated and found to express high levels of Notch1, Jagged1, Dll1 and Dll3 (Oktem et al 2014). CD133 and CD44 were both markers for cancer stem cells; therefore, it is believed that this subpopulation could be the putative prostate cancer stem cell (Oktem et al 2014).…”

“…In gene profiling of DU145 prostate cancer cells, a subpopulation with CD133 high /CD44 high was isolated and found to express high levels of Notch1, Jagged1, Dll1 and Dll3 (Oktem et al 2014). CD133 and CD44 were both markers for cancer stem cells; therefore, it is believed that this subpopulation could be the putative prostate cancer stem cell (Oktem et al 2014). In addition, it was reported that in human prostate cancer tissue samples, a subpopulation conferring docetaxel resistance, characterized by loss of epithelial differentiation markers and HLA class 1 (HLA1) antigens and marked up-regulation of Notch and Hedgehog signaling, was detected (Domingo-Domenech et al 2012).…”

Influence of CD133+ expression on patients' survival and resistance of CD133+ cells to anti-tumor reagents in gastric cancer

“…Various tumor precursor cells, especially of epithelial origin, highly express CD133, though tumor precursor cells in various tissues vary in phenotype [7][8][9][10][11][12][13][14][15][16][17]. The CD133 protein is located in the cell membrane projections and is involved in the formation of cell membrane topology.…”

“…It is, therefore, rational that human myeloblastic leukemia derives from stem cells [6]. TICs with CD133 + expression have been subsequently identified in various solid tumors, such as breast cancer, brain tumour, prostate cancer, malignant melanoma, colon cancer, liver cancer, pancreatic cancer and head and neck squamous cell carcinoma, though CD133 + cancerous cells have not been classified as cancer stem cells of stomach until now [7][8][9][10][11][12][13][14].…”

Lithium chloride has a biphasic effect on prostate cancer stem cells and a proportional effect on midkine levels

“…Cells in semi-confluent flasks (70% of confluency) were harvested using 0.05% trypsin (Sigma-Aldrich). Harvested cells were added to DMEM-F12 for trypsin inactivation and centrifuged (NF200; Nüve Laboratory and Sterilization Technology, Ankara, Turkey) at 500 x g for 3 min at 22˚C and then resuspended in culture medium (23).…”