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

Oktem,Gulperi; Bilir,Ayhan; Uslu,Ruchan; Inan,Sevinc V.; Demiray,Sirin B.; Atmaca,Harika; Ayla,Sule; Sercan,Ogun; Uysal,Aysegul

doi:10.3892/ol.2014.1992

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

Authors:
- Gulperi Oktem
- Ayhan Bilir
- Ruchan Uslu
- Sevinc V. Inan
- Sirin B. Demiray
- Harika Atmaca
- Sule Ayla
- Ogun Sercan
- Aysegul Uysal
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Affiliations: Department of Histology and Embryology, Faculty of Medicine, Ege University, Bornova, Izmir 35100, Turkey, Department of Histology and Embryology, Istanbul Medical Faculty, Istanbul University, Capa, Istanbul 34098, Turkey, Department of Medical Oncology, Faculty of Medicine, Ege University, Bornova, Izmir 35100, Turkey, Department of Histology and Embryology, Faculty of Medicine, Celal Bayar University, Manisa 45030, Turkey, Department of Biology, Faculty of Science and Art, Celal Bayar University, Manisa 45030, Turkey, Zeynep Kamil Gynecology and Maternity Training and Research Hospital, Istanbul 34668, Turkey, Department of Medical Biology, Faculty of Medicine, Dokuz Eylul University, Bornova, Izmir 35340, Turkey
Published online on: March 24, 2014 https://doi.org/10.3892/ol.2014.1992
Pages: 2103-2109

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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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