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. 2017 Jan;8(1-2):484-494.
doi: 10.18632/genesandcancer.133.

Genetically transforming human osteoblasts to sarcoma: development of an osteosarcoma model

Yi Yang  1   2 , Rui Yang  3 , Michael Roth  1 , Sajida Piperdi  1 , Wendong Zhang  1 , Howard Dorfman  3   4 , Pulivarthi Rao  5 , Amy Park  1 , Sandeep Tripathi  1 , Carrie Freeman  1 , Yunjia Zhang  1 , Rebecca Sowers  1 , Jeremy Rosenblum  1 , David Geller  3 , Bang Hoang  3 , Jonathan Gill  1 , Richard Gorlick  1   6   7
Affiliations

Genetically transforming human osteoblasts to sarcoma: development of an osteosarcoma model

Yi Yang et al. Genes Cancer. 2017 Jan.

Abstract

Osteosarcoma is the most common primary malignant bone tumor in children and young adults. Although histologically defined by the presence of malignant osteoid, the tumor possesses lineage multipotency suggesting it could be derived from a cell anywhere on the differentiation pathway between a mesenchymal stem cell (MSC) and a mature osteoblast. To determine if preosteoblasts (pOB) could be the cell of origin differentiated MSCs were transformed with defined genetic elements. MSCs and pOB differentiated from the same MSCs were serially transformed with the oncogenes hTERT, SV40 large T antigen and H-Ras. Assays were performed to determine their tumorigenic properties, differentiation capacity and histologic appearance. When subcutaneously implanted in immunocompromised mice, cell lines derived from transformed MSC and pOB formed tumors in 4 weeks. In contrast to the transformed MSC, the pOB tumors demonstrated a histological appearance characteristic of osteosarcoma. The cell lines derived from the transformed pOB only had osteogenic and chondrogenic differentiation potential, but not adipogenic ones. However, the transformed MSC cells and standard osteosarcoma cell lines maintained their tri-lineage differentiation capacity. The inability of the transformed pOB cell line to undergo adipogenic differentiation, may suggest that osteosarcoma is derived from a cell intermediate in differentiation between an MSC and a pOB, with partial commitment to the osteoblastic lineage.

Keywords: mesenchymal stem cells; osteoblast; osteosarcoma.

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Conflict of interest statement

CONFLICTS OF INTEREST The authors confirm that there are no conflicts of interest.

Figures

Figure 1
Figure 1. Anchorage independent growth in soft agar of transfected cell lines
Anchorage independent growth was measured in (A) hMSC-TSR; (B) pOB-TSR; (C) Colony numbers of MSC-TSR and pOB-TSR were significantly greater than that of mOB-TSR (P < .01).
Figure 2
Figure 2. Tumorigenicity assays in SCID mice
(A) Gross picture of hMSC-TSR and pOB-TSR cell lines implanted in mouse subcutaneously. (B) Tumors growth curve in mice after pOB-TSR cells injection. Once the size of the tumor reached to 1.7 cm in diameter or having 20% weight loss, the mouse was sacrificed according to the animal use protocol. (C-D) Histopathological findings of hMSC-TSR and pOB-TSR cell lines. Os, osteogenesis. T, tumor.
Figure 3
Figure 3. Changes in multilineage differentiation capacity in hMSC-TSR and pOB-TSR
Osteogenic differentiation staining with Alizarin Red, adipogenic differentiation staining with Oil Red O, and chondrogenic differentiation immunohistochemical staining with type II collagen.
Figure 4
Figure 4. Gene expression differences between transformed cell lines, untransformed cell lines, and osteosarcoma xenografts
(A) Unsupervised hierarchical clustering and heat map of gene expression correlation for untransformed cell lines, transformed cell lines, and osteosarcoma xenografts (M1, M9, M17, M31). Venn diagrams representing the number of genes differentially expressed (q-value <0.01) by transformed MSCs and pOBs as compared to (B) untransformed cell lines and (C) osteosarcoma xenografts. Red indicates increased expression and green indicates decreased expression.
Figure 5
Figure 5. Schematic representation for the use of tumorigenic MSCs and pOBs as a model to study osteosarcoma genesis
See this image and copyright information in PMC

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