Figure 1. LOC284454 is highly expressed in NPC and predicts unfavorable prognosis. (A) Differential gene expression ... Figure 1. LOC284454 is highly expressed.

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Figure 1. LOC284454 is highly expressed in NPC and predicts unfavorable prognosis. (A) Differential gene expression ... Figure 1. LOC284454 is highly expressed in NPC and predicts unfavorable prognosis. (A) Differential gene expression profile of top 20 altered lncRNAs in nasopharyngeal chronic inflammatory tissues and NPC tissues. (B) The expression of LOC284454 in gene expression profile (upper left). The expression of LOC284454 was measured in 9 nasopharyngeal chronic inflammatory tissues and 27 NPC tissues by RT-qPCR (upper right). Expression of LOC284454 in published GEO data of NPC (GSE53819 and GSE68799, lower panel). (C) LOC284454 expression was measured by ISH in non-tumor NPE (N) and NPC biopsies (T). Forty cases of non-tumor NPE (N) and 112 NPC biopsies with LOC284454 staining are shown. In paraffin sections, LOC284454 was highly expressed in NPC compared with that in adjacent non-tumor tissues. (D) OS and relapse-free survival were analyzed in patients with LOC284454 using a Kaplan–Meier curve. Patients with higher LOC284454 had a remarkable unfavorable prognosis compared with those with lower LOC284454. (E and F) The association between local relapse or distant metastasis and LOC284454. Unless provided in the caption above, the following copyright applies to the content of this slide: © The Author(s) 2018. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model) Carcinogenesis, Volume 40, Issue 2, 30 October 2018, Pages 380–391, https://doi.org/10.1093/carcin/bgy143 The content of this slide may be subject to copyright: please see the slide notes for details.

Figure 2. LOC284454 silencing suppresses tumor cell migration and invasion. (A) The expression of LOC284454 was ... Figure 2. LOC284454 silencing suppresses tumor cell migration and invasion. (A) The expression of LOC284454 was significantly increased in NPC cell lines (HNE1, HNE2, 5-8F and HK-1) compared with that in NP69, a normal human nasopharyngeal epithelial cell line used as a control. (B) The knockdown efficacy in 5-8F, HNE2 and HK-1 cell lines. The knockdown efficacy of siRNA1 + 2 was better than that of siRNA1 or siRNA2 alone, so we used siRNA1 + 2, referred to as siRNA, for subsequent knockdown. (C) Invasive capacity was largely impaired after transfection with LOC284454 siRNA compared with NC. (D) Wound healing assay was conducted to measure the migration potential. Images were captured at different time points according to the actual healing situation. The migration potential was limited in the LOC284454 siRNA group compared with NC. Unless provided in the caption above, the following copyright applies to the content of this slide: © The Author(s) 2018. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model) Carcinogenesis, Volume 40, Issue 2, 30 October 2018, Pages 380–391, https://doi.org/10.1093/carcin/bgy143 The content of this slide may be subject to copyright: please see the slide notes for details.

Figure 3. LOC284454 overexpression promotes tumor cell migration and invasion. (A) Transfection of LOC284454 ... Figure 3. LOC284454 overexpression promotes tumor cell migration and invasion. (A) Transfection of LOC284454 overexpression plasmid (OE) or empty vector was evaluated in HNE2, 5-8F and HK-1 cells, and LOC284454 was significantly overexpressed. (B) After transfection with the LOC284454-overexpressed plasmid, the Matrigel invasion ability was increased compared with transfection with empty vector. (C) Enhanced wound healing capacity observed in the LOC284454-overexpressed group compared with the empty vector group. Unless provided in the caption above, the following copyright applies to the content of this slide: © The Author(s) 2018. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model) Carcinogenesis, Volume 40, Issue 2, 30 October 2018, Pages 380–391, https://doi.org/10.1093/carcin/bgy143 The content of this slide may be subject to copyright: please see the slide notes for details.

Figure 4. LOC284454 increases lung metastasis in vivo Figure 4. LOC284454 increases lung metastasis in vivo. (A) Image of fresh lungs dissected from each group of mice ... Figure 4. LOC284454 increases lung metastasis in vivo. (A) Image of fresh lungs dissected from each group of mice treated with LOC284454-overexpressed, LOC284454-siRNA or NC 5-8F cells (seven mice in each group). (B) Metastases visible on mouse lung are shown. The white dots are nodules, and the arrows indicate colonies formed by the tumor cells on the lungs. (C) Quantities of metastases in each group, with each dot representing one mouse. (D) Mouse lung tissue stained with hematoxylin and eosin. Rectangular frame indicates amplified part of metastatic tumor cells in the lung. The images were taken at ×100 and ×400. Unless provided in the caption above, the following copyright applies to the content of this slide: © The Author(s) 2018. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model) Carcinogenesis, Volume 40, Issue 2, 30 October 2018, Pages 380–391, https://doi.org/10.1093/carcin/bgy143 The content of this slide may be subject to copyright: please see the slide notes for details.

Figure 5. Identification of LOC284454 regulated partners with LC-MS/MS combined with bioinformatics. (A) LC-MS/MS data ... Figure 5. Identification of LOC284454 regulated partners with LC-MS/MS combined with bioinformatics. (A) LC-MS/MS data analysis process. The number of proteins obtained from NC, siRNA, vector and overexpression were 1624, 1604, 1494 and 1476, respectively. The data were normalized to remove systematic errors. After filtering out the area of protein <1.0E5, the ratio of si/NC <0.67 and OE/vector >1.5 or vice versa was considered significant. By intersecting the siRNA group and overexpression group, 193 upregulated and 60 downregulated proteins were identified. (B) Expression of Rho/Rac pathway-associated proteins. Dysregulated protein levels were detected by western blot in 5-8F, HNE2 and HK1 cells transfected with NC, LOC284454 siRNA, empty vector or LOC284454 overexpression plasmid (OE). GAPDH was used as an endogenous control. Unless provided in the caption above, the following copyright applies to the content of this slide: © The Author(s) 2018. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model) Carcinogenesis, Volume 40, Issue 2, 30 October 2018, Pages 380–391, https://doi.org/10.1093/carcin/bgy143 The content of this slide may be subject to copyright: please see the slide notes for details.

Figure 6. Proposed schematic model illustrating the role of LOC284454 in regulating NPC migration and invasion by ... Figure 6. Proposed schematic model illustrating the role of LOC284454 in regulating NPC migration and invasion by inducing Rho/Rac signaling pathway changes. LOC284454 influences the Rho/Rac signaling pathway by promoting or inhibiting the proteins associated with the cytoskeleton and cell adhesion, inducing actin polymerization, remodeling of the cytoskeleton and changes in cell adhesion, which further affect the migration and invasion of NPC cells. Unless provided in the caption above, the following copyright applies to the content of this slide: © The Author(s) 2018. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model) Carcinogenesis, Volume 40, Issue 2, 30 October 2018, Pages 380–391, https://doi.org/10.1093/carcin/bgy143 The content of this slide may be subject to copyright: please see the slide notes for details.