title: Generation of Osteosarcomas from a Combination of Rb Silencing and c-Myc Overexpression in Human Mesenchymal Stem Cells.
abstract: Osteosarcoma (OS) was a malignant tumor occurring with unknown etiology that made prevention and early diagnosis difficult. Mesenchymal stem cells (MSCs), which were found in bone marrow, were claimed to be a possible origin of OS but with little direct evidence. We aimed to characterize OS cells transformed from human MSCs (hMSCs) and identify their association with human primary OS cells and patient survival. Genetic modification with p53 or retinoblastoma (Rb) knockdown and c-Myc or Ras overexpression was applied for hMSC transformation. Transformed cells were assayed for proliferation, differentiation, tumorigenecity, and gene expression profile. Only the combination of Rb knockdown and c-Myc overexpression successfully transformed hMSCs derived from four individual donors, with increasing cell proliferation, decreasing cell senescence rate, and increasing ability to form colonies and spheres in serum-free medium. These transformed cells lost the expression of certain surface markers, increased in osteogenic potential, and decreased in adipogenic potential. After injection in immunodeficient mice, these cells formed OS-like tumors, as evidenced by radiographic analyses and immunohistochemistry of various OS markers. Microarray with cluster analysis revealed that these transformed cells have gene profiles more similar to patient-derived primary OS cells than their normal MSC counterparts. Most importantly, comparison of OS patient tumor samples revealed that a combination of Rb loss and c-Myc overexpression correlated with a decrease in patient survival. This study successfully transformed human MSCs to OS-like cells by Rb knockdown and c-Myc overexpression that may be a useful platform for further investigation of preventive and target therapy for human OS. Stem Cells Translational Medicine 2017;6:512-526.
authors.affiliation: Department of Orthopaedics and Traumatology, Taipei Veterans General Hospital, Taipei, Taiwan, Republic of China.|Department of Orthopaedics, Therapeutical and Research Center of Musculoskeletal Tumor, Taipei Veterans General Hospital, Taipei, Taiwan, Republic of China.|Institute of Traditional Medicine, School of Medicine, National Yang-Ming University, Taipei, Taiwan, Republic of China.
authors.forename: Jir-You
authors.initials: JY
authors.lastname: Wang
authors.affiliation: Department of Orthopaedics and Traumatology, Taipei Veterans General Hospital, Taipei, Taiwan, Republic of China.|Department of Orthopaedics, Therapeutical and Research Center of Musculoskeletal Tumor, Taipei Veterans General Hospital, Taipei, Taiwan, Republic of China.
authors.forename: Po-Kuei
authors.initials: PK
authors.lastname: Wu
authors.affiliation: Department of Orthopaedics, Therapeutical and Research Center of Musculoskeletal Tumor, Taipei Veterans General Hospital, Taipei, Taiwan, Republic of China.|Department of Pathology and Laboratory Medicine, Taipei Veterans General Hospital, Taipei, Taiwan, Republic of China.
authors.forename: Paul Chih-Hsueh
authors.initials: PC
authors.lastname: Chen
authors.affiliation: Department of Orthopaedics and Traumatology, Taipei Veterans General Hospital, Taipei, Taiwan, Republic of China.|Department of Orthopaedics, Therapeutical and Research Center of Musculoskeletal Tumor, Taipei Veterans General Hospital, Taipei, Taiwan, Republic of China.
authors.forename: Chia-Wen
authors.initials: CW
authors.lastname: Lee
authors.affiliation: Department of Orthopaedics and Traumatology, Taipei Veterans General Hospital, Taipei, Taiwan, Republic of China.|Department of Orthopaedics, Therapeutical and Research Center of Musculoskeletal Tumor, Taipei Veterans General Hospital, Taipei, Taiwan, Republic of China.
authors.forename: Wei-Ming
authors.initials: WM
authors.lastname: Chen
authors.affiliation: Department of Orthopaedics and Traumatology, Taipei Veterans General Hospital, Taipei, Taiwan, Republic of China.|Department of Orthopaedics, Therapeutical and Research Center of Musculoskeletal Tumor, Taipei Veterans General Hospital, Taipei, Taiwan, Republic of China.|Institute of Traditional Medicine, School of Medicine, National Yang-Ming University, Taipei, Taiwan, Republic of China.|Institute of Clinical Medicine, School of Medicine, National Yang-Ming University, Taipei, Taiwan, Republic of China.|Department of Pharmacology, School of Medicine, National Yang-Ming University, Taipei, Taiwan, Republic of China.|Medical Research, Taipei Veterans General Hospital, Taipei, Taiwan, Republic of China.|Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan, Republic of China.|Integrative Stem Cell Center, China Medical University Hospital, Taichung, Taiwan, Republic of China.|Graduate Institute of Clinical Medical Science, China Medical University, Taichung, Taiwan, Republic of China.
authors.forename: Shih-Chieh
authors.initials: SC
authors.lastname: Hung
mesh_terms: D050156:Adipogenesis; D000818:Animals; D001859:Bone Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D049109:Cell Proliferation; D002471:Cell Transformation, Neoplastic / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D016922:Cellular Senescence; D015972:Gene Expression Regulation, Neoplastic; D020868:Gene Silencing; D006801:Humans; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000378:metabolism* / Q000473:pathology; D008819:Mice, Nude; D010012:Osteogenesis; D012516:Osteosarcoma / Q000235:genetics / Q000378:metabolism* / Q000556:secondary; D010641:Phenotype; D061251:Primary Cell Culture; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics* / Q000378:metabolism*; D016160:Retinoblastoma Protein / Q000172:deficiency* / Q000235:genetics; D013997:Time Factors; D059467:Transcriptome; D047368:Tumor Burden; D014407:Tumor Cells, Cultured; D015854:Up-Regulation
chemical_list: C489427:MYC protein, human; D016271:Proto-Oncogene Proteins c-myc; D016160:Retinoblastoma Protein
keywords: Mesenchymal stem cell; Osteosarcoma; Retinoblastoma (Rb) knockdown; Transformation; c-Myc