Background: Epstein-Barr virus associated gastric cancer (EBVaGC) become a growing health problem. TP73-AS1 showed high expression in EBVaGC cells. However, the function role and underlying mechanism of TP73-AS1 need further exploration.
Methods: The expressions of TP73-AS1, WIF1, EZH2, β-catenin and epithelial-mesenchymal transition (EMT)-related proteins were detected using qRT-PCR and Western blotting. Cell proliferation, apoptosis, migration and invasion were measured by CCK-8, colony formation, flow cytometry, wound healing and transwell assays, respectively. WIF1 promoter methylation was analyzed by MS-PCR (MSP). RNA immunoprecipitation assay (RIP) and Chromatin immunoprecipitation assay (ChIP) measured the interactions of TP73-AS1/EZH2 and EZH2/WIF1. Subcutaneous tumor growth was monitored in nude mice and immunohistochemistry (IHC) detected proliferation marker Ki-67 expression.
Results: TP73-AS1 was increased while WIF1 was decreased in EBVaGC cells. Silencing of TP73-AS1 or overexpression of WIF1 repressed the growth and migration while promoted apoptosis of EBVaGC cells. Knockdown of WIF1 reversed the anticancer effect of TP73-AS1 silencing. TP73-AS1 promoted the binding of EZH2 to the WIF1 promoter by directly binding to EZH2, and thus inhibiting the expression of WIF1 by enhancing H3K27me3 level of WIF1 promoter. Moreover, TP73-AS1 activated Wnt/β-catenin signaling pathway and promoted EMT by down-regulating WIF1. TP73-AS1 silencing inhibited the progression of EBVaGC in nude mice by epigenetically regulating WIF1.
Conclusion: TP73-AS1 regulated the promoter methylation of WIF1 by recruiting PRC2 complex to WIF1 promoter region, thereby promoting the progression of EBVaGC. These observations provided a novel theoretical basis to investigate more effective therapies of EBVaGC.
Keywords: EZH2; Epstein-Barr virus associated gastric cancer; LncRNA TP73-AS1; Promoter methylation; WIF1.
Copyright © 2018. Published by Elsevier Inc.