Adenovirus-mediated TIPE2 overexpression inhibits gastric cancer metastasis via reversal of epithelial-mesenchymal transition.

Tumor necrosis factor (TNF)-alpha-induced protein 8-like 2 (TNFAIP8L2; also termed TIPE2) has been shown to be involved in both the immune-negative modulation and cancer. We previously found that TIPE2 is lost in human gastric cancer, and TIPE2 restoration suppresses gastric cancer growth by induction of apoptosis and impairment of protein kinase B (PKB/AKT) and extracellular signal-regulated kinase-1/2 (ERK1/2) signaling. However, its correlation with epithelial-mesenchymal transition (EMT) in gastric cancer is largely elusive. In the present report, we carried out a gain-of-function study in AGS and HGC-27 human gastric cancer cells by adenovirus-mediated human TIPE2 gene transfer (AdVTIPE2). We then examined the effects of AdVTIPE2 on in vitro migration and invasion of AGS and HGC-27 tumor cells by wound-healing assay and Transwell invasion assay, respectively. We also investigated the effects of AdVTIPE2 on in vivo lung metastasis of AGS and HGC-27 tumor cells by intravenous (i.v.) injection in athymic BALB/c nude mice. We demonstrated that AdVTIPE2 remarkably suppressed the migratory, invasive and metastatic potential of AGS and HGC-27 tumor cells in vitro and in vivo in BALB/c nude mouse model. Mechanistically, AdVTIPE2 obviously upregulated E-cadherin epithelial marker in AGS and HGC-27 tumor cells, whereas it downregulated N-cadherin and Vimentin mesenchymal markers, Snail1, Snail2/Slug and Zeb1 EMT-inducing transcription factors (EMT-TFs), and tripartite motif-containing 29 (TRIM29) and phosphatase regenerating liver 3 (PRL-3) gastric cancer-specific metastasis markers. Importantly, glycogen synthase kinase-3β (GSK-3β) inhibitor VIII and 26S proteasome inhibitor MG132 assays revealed that TIPE2 downregulated Snail1 and Snail2/Slug in a GSK-3β- and proteasome-dependent manner possibly by impairing AKT signaling. Our data provided the first evidence that TIPE2 inhibits gastric cancer cell migration, invasion and metastasis very probably via reversal of EMT, revealing that TIPE2 may be a novel therapeutic target for human gastric cancer EMT and metastasis.