Acquisition of cancer stem cell properties in osteosarcoma cells by defined factors

• Main Authors: Shuichi Fujiwara, Teruya Kawamoto, Yohei Kawakami, Yasufumi Koterazawa, Hitomi Hara, Toshiyuki Takemori, Kazumichi Kitayama, Shunsuke Yahiro, Kenichiro Kakutani, Tomoyuki Matsumoto, Takehiko Matsushita, Takahiro Niikura, Michiyo Koyanagi-Aoi, Takashi Aoi, Ryosuke Kuroda, Toshihiro Akisue
• Format: Article
• Language: English
• Published: BMC 2020-10-01
• Series: Stem Cell Research & Therapy
• Subjects: Cancer stem cells; Chemoresistance; Osteosarcoma; Sarcosphere; Tumorigenicity
• Online Access: http://link.springer.com/article/10.1186/s13287-020-01944-9

Abstract Background Cancer stem cells (CSCs) are considered to be responsible for tumor initiation, formation, and poor prognosis of cancer patients. However, the rarity of CSCs in clinical samples makes it difficult to elucidate characteristics of CSCs, especially in osteosarcoma (OS). The aim of this study is to verify whether it is possible to generate CSC-like cells by transducing defined factors into an OS cell line. Methods We retrovirally transduced the Octamer-binding transcription factor 3/4 (OCT3/4), Kruppel-like factor 4 (KLF4), and SRY-box transcription factor 2 (SOX2) genes into the MG-63 human OS cell line (MG-OKS). Parental and GFP-transduced MG-63 cells were used as negative control. We assessed the properties of the generated cells in vitro and in vivo. Multiple comparisons among groups were made using a one-way analysis of variance (ANOVA) followed by post hoc testing with Tukey’s procedure. Results MG-OKS cells in vitro exhibited the significantly increased mRNA expression levels of CSC markers (CD24, CD26, and CD133), decreased cell growth, increased chemoresistance and cell migration, and enhanced sphere formation. Notably, MG-OKS cells cultured under osteogenic differentiation conditions showed strongly positive staining for both Alizarin Red S and alkaline phosphatase, indicating osteogenesis of the cells. Gene ontology analysis of microarray data revealed significant upregulation of epidermal-related genes. Tumors derived from MG-OKS cells in vivo were significantly larger than those from other cells in μCT analysis, and immunohistochemical staining showed that Ki-67, osteocalcin, and HIF-1α-positive cells were more frequently detected in the MG-OKS-derived tumors. Conclusions In this study, we successfully generated OS CSC-like cells with significantly enhanced CSC properties following transduction of defined factors.