title: Reprogramming of retinoblastoma cancer cells into cancer stem cells.
abstract: Retinoblastoma is the most common intraocular malignancy in pediatric patients. It develops rapidly in the retina and can be fatal if not treated promptly. It has been proposed that a small population of cancer cells, termed cancer stem cells (CSCs), initiate tumorigenesis from immature tissue stem cells or progenitor cells. Reprogramming technology, which can convert mature cells into pluripotent stem cells (iPS), provides the possibility of transducing malignant cancer cells back to CSCs, a type of early stage of cancer. We herein took advantage of reprogramming technology to induce CSCs from retinoblastoma cancer cells. In the present study, the 4 Yamanaka transcription factors, Oct4, Sox2, Klf4 and c-myc, were transduced into retinoblastoma cells (Rbc51). iPS-like colonies were observed 15 days after transduction and showed significantly enhanced CSC properties. The gene and protein expression levels of pluripotent stem cell markers (Tra-1-60, Oct4, Nanog) and cancer stem cell markers (CD133, CD44) were up-regulated in transduced Rbc51 cells compared to control cells. Moreover, iPS-like CSCs could be sorted using the Magnetic-activated cell sorting (MACS) method. A sphere formation assay demonstrated spheroid formation in transduced Rbc51 cells cultured in serum free media, and these spheroids could be differentiated into Pax6-, Nestin-positive neural progenitors and rhodopsin- and recoverin-positive mature retinal cells. The cell viability after 5-Fu exposure was higher in transduced Rbc51 cells. In conclusion, CSCs were generated from retinoblastoma cancer cells using reprogramming technology. Our novel method can generate CSCs, the study of which can lead to better understanding of cancer-specific initiation, cancer epigenetics, and the overlapping mechanisms of cancer development and pluripotent stem cell behavior.
authors.affiliation: Department of Histology and Embryology, Shinshu University School of Medicine, 3-1-1 Asahi, Matsumoto 390-8621, Japan. Electronic address: yuefm75@gmail.com.
authors.forename: Fengming
authors.initials: F
authors.lastname: Yue
authors.affiliation: Department of Histology and Embryology, Shinshu University School of Medicine, 3-1-1 Asahi, Matsumoto 390-8621, Japan.
authors.forename: Kanji
authors.initials: K
authors.lastname: Hirashima
authors.affiliation: Department of Biotechnology and Biomedical Engineering, Institute for Biomedical Sciences, Interdisciplinary Cluster for Cutting Edge Research, Shinshu University Matsumoto, 3-1-1 Asahi, Matsumoto 390-8621, Japan.
authors.forename: Daihachiro
authors.initials: D
authors.lastname: Tomotsune
authors.affiliation: Bourbon Corporation, 4-2-14 Matsunami, Kashiwazaki, Niigata 945-8611, Japan.
authors.forename: Sakiko
authors.initials: S
authors.lastname: Takizawa-Shirasawa
authors.affiliation: Bourbon Corporation, 4-2-14 Matsunami, Kashiwazaki, Niigata 945-8611, Japan.
authors.forename: Tadayuki
authors.initials: T
authors.lastname: Yokoyama
authors.affiliation: Department of Histology and Embryology, Shinshu University School of Medicine, 3-1-1 Asahi, Matsumoto 390-8621, Japan; Department of Biotechnology and Biomedical Engineering, Institute for Biomedical Sciences, Interdisciplinary Cluster for Cutting Edge Research, Shinshu University Matsumoto, 3-1-1 Asahi, Matsumoto 390-8621, Japan.
authors.forename: Katsunori
authors.initials: K
authors.lastname: Sasaki
mesh_terms: D045744:Cell Line, Tumor; D065150:Cellular Reprogramming; D000067470:Cellular Reprogramming Techniques / Q000379:methods*; D018507:Gene Expression Regulation, Developmental; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D000090062:Kruppel-Like Factor 4; D014411:Neoplastic Stem Cells / Q000166:cytology* / Q000378:metabolism; D012160:Retina / Q000166:cytology / Q000378:metabolism; D019572:Retinal Neoplasms / Q000235:genetics*; D012175:Retinoblastoma / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics; D014161:Transduction, Genetic
chemical_list: C000715467:KLF4 protein, human; D000090062:Kruppel-Like Factor 4; D014157:Transcription Factors
keywords: Cancer cells; Cancer initiation mechanism; Cancer stem cells (CSCs); Pluripotent stem cells; Reprogramming; Retinoblastoma