Acquisition of cancer stem cell properties in osteosarcoma cells by defined factors
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(2020) 11:429
RESEARCH
Open Access
Acquisition of cancer stem cell properties in osteosarcoma cells by defined factors Shuichi Fujiwara1, Teruya Kawamoto1,2* , Yohei Kawakami1, Yasufumi Koterazawa3,4,5, Hitomi Hara1, Toshiyuki Takemori1, Kazumichi Kitayama1, Shunsuke Yahiro1, Kenichiro Kakutani1, Tomoyuki Matsumoto1, Takehiko Matsushita1, Takahiro Niikura1, Michiyo Koyanagi-Aoi3,4,6, Takashi Aoi3,4,6, Ryosuke Kuroda1 and Toshihiro Akisue1,7
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 MGOKS 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-OKSderived tumors. Conclusions: In this study, we successfully generated OS CSC-like cells with significantly enhanced CSC properties following transduction of defined factors. Keywords: Cancer stem cells, Chemoresistance, Osteosarcoma, Sarcosphere, Tumorigenicity
* Correspondence: [email protected] 1 Department of Orthopaedic Surgery, Kobe University Graduate School of Medicine, 7-5-1 Kusunoki-cho, Chuo-ku, Kobe 650-0017, Japan 2 Division of Orthopaedic Surgery, Kobe University Hospital International Clinical Cancer Research Center, Kobe, Japan Full list of author information is available at the end of the article © The Author(s). 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the
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