Abstract
Purpose
Osteosarcoma (OS) is one of the most prevalent primary malignant bone tumors in adolescent. HOTAIR is highly expressed and associated with the epigenetic modifications, especially DNA methylation, in cancer. However, the regulation mechanism between HOTAIR and DNA methylation and the biological effects of them in the pathogenesis of osteosarcoma remains elusive.
Method
Through RNA-sequencing and computational analysis, followed by a variety of experimental validations, we report a novel interplay between HOTAIR, miR-126, and DNA methylation in OS.
Results
We found that HOTAIR is highly expressed in OS cells and the knockdown of HOTAIR leads to the down-regulation of DNMT1, as well as the decrease of global DNA methylation level. RNA-sequencing analysis of HOTAIR-regulated gene shows that CDKN2A is significantly repressed by HOTAIR. A series of experiments show that HOTAIR represses the expression of CDKN2A through inhibiting the promoter activity of CDKN2A by DNA hypermethylation. Further evidence shows that HOTAIR activates the expression of DNMT1 through repressing miR-126, which is the negative regulator of DNMT1. Functionally, HOTAIR depletion increases the sensibility of OS cells to DNMT1 inhibitor through regulating the viability and apoptosis of OS cells via HOTAIR-miR126-DNMT1-CDKN2A axis.
Conclusion
These results not only enrich our understanding of the regulation relationship between non-coding RNA, DNA methylation, and gene expression, however, also provide a novel direction in developing more sophisticated therapeutic strategies for OS patients.
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Abbreviations
- OS:
-
Osteosarcoma
- MDS:
-
Myelodysplastic syndrome
- DAC:
-
Decitabine
- RT:
-
Reverse transcription
- ChIP:
-
Chromatin immunoprecipitation
- qPCR:
-
Quantitative PCR
- MedIP:
-
Methylated DNA immunoprecipitation
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FS and XL designed the study; XL, HL, GF, MH, YS, and KX performed experiments; YL and HL performed data analysis; XL and FS wrote the manuscript; FS, XL, and HL revised the manuscript; and all authors approved the manuscript.
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All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.
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Informed consent was obtained from all individual participants included in the study.
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Xingang Li and Hongming Lu are co-first authors.
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Li, X., Lu, H., Fan, G. et al. A novel interplay between HOTAIR and DNA methylation in osteosarcoma cells indicates a new therapeutic strategy. J Cancer Res Clin Oncol 143, 2189–2200 (2017). https://doi.org/10.1007/s00432-017-2478-3
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DOI: https://doi.org/10.1007/s00432-017-2478-3