Human glioma-initiating cells stably transfected with red fluorescent protein gene can spontaneously fuse with host-derived bone marrow mesenchymal stem cells and induce malignant transformation

doi:10.3969/j.issn.2095-4344.1782

Abstract

Abstract:

BACKGROUND: Malignant transformation of host-derived stromal cells in xenograft tumors has been reported, but little is known about the mechanisms of malignant transformation.

OBJECTIVE: To investigate the malignant transformation of host-derived tumor stromal cells and to explore the possible mechanisms in an in vivo model of two-color fluorescent tracing.

METHODS: Human glioma-initiating cells stably transfected with red fluorescent protein gene (GICs-RFP) were inoculated into nude mice expressing enhanced green fluorescent protein (EGFP). An in vivo model of two-color fluorescence tracer was established. The fluorescent expression of transplanted tumors was observed by confocal laser scanning. Flow cytometry was used to detect and classify various fluorescent cells, including EGFP cells, RFP cells and EGFP/RFP fusion cells. The fusion cells co-expressed EGFP/RFP and had malignant proliferation characteristics because of in vitro subcloning. Origin, surface markers and malignant features of the cells were identified in vitro. The tumorigenicity of fusion cells was verified by subcutaneous transplantation in nude mice. The study was approved by the Animal Ethics Committee of Soochow University (approval No. ECSU-201800090).

RESULTS AND CONCLUSION: The tumorigenic rates of GICs-RFP in EGFP nude mice were 100% (14/14). Fusion cells were observed in the transplanted tumors of all the animal models. The fusion cells not only co-expressed EGFP and RFP, but also co-expressed GICs-RFP marker Nestin and bone marrow mesenchymal stem cell markers CD44, CD105 and CD29. Fusion cells showed high proliferative activity and more invasive and migratory characteristics in vitro as compared with GICs-RFP. The tumorigenicity of fusion cells (1×10⁵ cells per mouse) in thymus-free nude mice was 100% (4/4). These results indicate that GICs-RFP spontaneously fuses with host-derived bone marrow mesenchymal stem cells and induces malignant transformation, which provides new evidence for two-color fluorescence tracing of heterogeneous tumor cell sources.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: fluorescent protein tracing, cell fusion, malignant transformation, glioma-initiating cells, fluorescent nude mice, tumor microenvironment, animal model, mesenchymal stem cells, tumor stem cells

CLC Number:

R441
R364

Han Hui, Dai Xingliang, Cheng Hongwei, Lan Qing. Human glioma-initiating cells stably transfected with red fluorescent protein gene can spontaneously fuse with host-derived bone marrow mesenchymal stem cells and induce malignant transformation[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(25): 3944-3950.

Figures/Tables 7

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