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. 2018 Sep 24;8:14294. doi: 10.1038/s41598-018-32428-9

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© The Author(s) 2018

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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Skp2 knockdown inhibits tumor growth and lung metastasis in an orthotopic osteosarcoma model. (A) Experimental design. 5 × 10⁵ 143B/shctrl or 143B/shSkp2 cells were implanted into the proximal tibias of mice. Legs were amputated once any tumor reached 1.5 cm in diameter. Animals were kept for 2 weeks after amputation and then sacrificed. (B) Representative pictures of tibia tumors. (C) Tumor volumes of the Skp2-knockdown group were significantly than the control group. (D) Representative pictures of lungs harvested from mice injected with transfected 143B/shctrl and 143B/shSkp2 cell lines. (E) The mean lung weight (LW) normalized to body weight (BW) was significantly greater in the control group compared to the Skp2-knockdown group. (F) Skp2 knockdown significantly reduced the number of metastatic lung nodules. (G) Expression of Skp2 from two different lungs was examined by Western blot. No Skp2 expression was observed in143B/shSkp2 lung tissue. (H) Representative pictures of hematoxylin-eosin (HE) and IHC staining of lung tissues. Lower levels of Skp2 immunostaining were detected in metastatic lung nodules from the 143B/shSkp2 group. Statistical significance is indicated by: *p < 0.05, **p < 0.01, ***p < 0.001. Column: mean; Error bars: SD.