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. 2017 Apr 3;36(31):4405–4414. doi: 10.1038/onc.2017.69

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Copyright © 2017 The Author(s)

This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/4.0/

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Coexpression of SPZ1 and TWIST1 transgenes promotes EMT and invasive activity. (a) Effect of forced coexpression of SPZ1 and TWIST1 introduced into Hep G2 and Hep 3B cells on expression of mesenchymal markers. a, P <0.001 and b, P<0.01. (b) Effect of combination of knockdown siRNA constructs of SPZ1 and TWIST1 introduced into SK-Hep1 cells on expression of mesenchymal markers. Upper panel, mRNA levels; lower panel, protein levels. a, P<0.001 and b, P<0.01. (c) Effect of TWIST1 siRNA on Tet-ON inducible SPZ1-GFP on expression of mesenchymal marker proteins in SK-Hep1 and Huh 7 cells. (d) Analysis of wound healing in Huh 7 transfected with SPZ1 and/or TWIST1 expression vectors as described in the Materials and methods section. (e) Coexpression of SPZ1-GFP and TWIST1 promoted invasive activity in Huh 7. Upper panels, hematoxin staining; lower panels, relative activity of invasion activity. (f) Analysis of wound healing in SPZ1 or TWIST1 shRNAi1-transfected SK-Hep1, as described in the Materials and methods section. (g) SK-Hep1 with SPZ1 and TWIST1 knockdown decreased invasive activity compared with vector-transfected cells. (h) SPZ1 shRNAi1- and TWIST1 shRNAi-transfected SK-Hep1 cells abolished the invasive activity in vivo (n=5). Upper panel, images of inoculated recombinant tumor cells; lower panel, comparison of the relative photon flux activity. All except (a) and (b) are indicated as a and b, P<0.001.