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. 2019 Dec 30;9:20360. doi: 10.1038/s41598-019-56883-0

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Effect of DNA demethylation therapy against OS cell lines. (A) Experimental protocol of drug treatment in vitro. Four OS cell lines (MG63, HOS, 143B, MNNG/HOS) were treated with 5-aza-dC (0, 0.1, 0.3, 1, 3, and 10 μM) for 4 days, and the cell number was counted on day 9. Cell growth was significantly suppressed by treatment with low doses of 5-aza-dC (0.1–0.3 μM). (B–D) The in vivo effect of the DNA methylation therapy. 5-Aza-dC was intraperitoneally administered for 3 days followed by drug holidays of 4 days, and the treatment was repeated for three or four cycles (top of C, and D). The treatment with 5-aza-dC reduced tumor volume (B–D) and weight (B) of both MNNG/HOS and 143B xenograft tumors. (E,F) Pathological analysis of the xenografts. In the 5-aza-dC-treated tumors, marked tumor cell necrosis was observed (E,F). *P < 0.05, **P < 0.01, ***P < 0.001.