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Bone mesenchymal stem cell-derived extracellular vesicles containing NORAD promote osteosarcoma by miR-30c-5p

Laboratory Investigation volume 102, pages 826–837 (2022)Cite this article

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A Correction to this article was published on 19 April 2022

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Abstract

Osteosarcoma is a bone tumor that often affects children, adolescents and young people. Non-coding RNA activated by DNA damage (NORAD) can promote the proliferation of cancer cells in multiple tumors. Thus, the current study set out to explore the role of NORAD derived from extracellular vesicles (EVs) of bone mesenchymal stem cells (BMSCs) in osteosarcoma. First, NORAD was highly expressed in osteosarcoma cells and tissues, which might be associated with the progression and metastasis of osteosarcoma. We isolated EVs from the characterized BMSCs, and found that NORAD was transferred from BMSCs to osteosarcoma cells via EVs in the co-culture system. Consequently, NORAD delivered by BMSC-derived EVs promoted the proliferation and invasion of osteosarcoma cells. Subsequently, bioinformatics analyses suggested potential binding relationship between NORAD and microRNA-30c-5p (miR-30c-5p) as well as between miR-30c-5p and Krueppel-like factor 10 (KLF10), and the results of which were further verified by dual luciferase reporter gene assay, RNA immunoprecipitation, and RNA pull-down assay. Mechanistically, NORAD acted as a sponge of miR-30c-5p and up-regulated the expression of KLF10 where miR-30-c-5p mimic declined the effect induced by NORAD on cancer cells. The osteosarcoma cells were injected into mice to develop tumor growth and metastasis models. In these two models, injection of BMSC-EVs elevated NORAD expression and KLF10 but reduced miR-30c-5p expression, whereby suppressing tumor growth and lung metastasis. To conclude, BMSC-EVs deliver NORAD to osteosarcoma cells to regulate the miR-30c-5p/KLF10 axis, thereby accelerating the progression and metastasis of osteosarcoma.

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Fig. 1: NORAD induced lung metastasis of osteosarcoma.
Fig. 2: BMSC-EVs transmitted NORAD to osteosarcoma cells.
Fig. 3: NORAD in BMSC-EVs enhanced the proliferative and invasive potentials of osteosarcoma cells.
Fig. 4: NORAD adsorbing miR-30c-5p affected the proliferation and invasion of osteosarcoma cells.
Fig. 5: NORAD promoted KLF10 expression via sponging miR-30c-5p.
Fig. 6: EV-NORAD promoted the proliferation and invasion of osteosarcoma cells via miR-30c-5p/KLF10 axis.
Fig. 7: NORAD from BMSC-EVs promoted lung metastasis of osteosarcoma.
Fig. 8: BMSC-EVs transferred NORAD to osteosarcoma cells.

Availability of data and materials

The data that support the findings of this study are available from the corresponding authors upon reasonable request.

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Authors and Affiliations

  1. Department of Traumatic Orthopedics, The Second Hospital of Dalian Medical University, Dalian, 116023, China

    Hongtao He, Wenzhi Zhao, Lu Zhang & Peng Yin

  2. Department of Orthopedics, The Second Xiangya Hospital, Central South University, Changsha, 410011, China

    Muliang Ding

  3. Department of Orthopedics, Renji Hospital Affiliated to Shanghai Jiaotong University School of Medicine, Shanghai, 200092, China

    Tao Li

  4. Department of Spine Surgery, The Second Hospital of Dalian Medical University, Dalian, 116023, China

    Wei Zhang

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Contributions

Conceived and designed research: HH, WZ, PY; Performed experiments: HH, MD, TL; Analyzed data: WZ; Interpreted results of experiments: HH, MD, TL; Prepared figures: LZ; Drafted paper: HH, MD, TL, WZ; Edited and revised paper: LZ, WZ, PY; Approved final version of paper: HH, MD, TL, WZ, LZ, WZ, PY.

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Correspondence to Wei Zhang.

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He, H., Ding, M., Li, T. et al. Bone mesenchymal stem cell-derived extracellular vesicles containing NORAD promote osteosarcoma by miR-30c-5p. Lab Invest 102, 826–837 (2022). https://doi.org/10.1038/s41374-021-00691-6

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