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Overexpression of cancer upregulated gene 2 (Cug2) decreases spry2 through c-cbl, leading to activation of egfr and β-catenin signaling
Author(s)
Yawut, Natpaphan
Kaewpiboon, Chutima
Budluang, Phatcharaporn
Cho, Il Rae
Kaowinn, Sirichat
Koh, Sang Seok
Chung, Young Hwa
Date Issued
January 1, 2020
Type
Article
Abstract
Purpose: The mechanism by which cancer upregulated gene 2 (CUG2) overexpression induces cancer stem cell-like phenotypes is not fully understood. Because the increased activity and expression of epidermal growth factor receptor (EGFR) kinase have been reported in A549 cancer cells overexpressing CUG2 (A549-CUG2) compared with control cells (A549-Vec), the Sprouty2 (Spry2) protein has gained attention as the downstream molecule of EGFR signaling. Therefore, we aim to identify the role of Spry2 in CUG2overexpressing lung cancer cells. Materials and Methods: Spry2 expression levels were examined in A549-CUG2 and A549-Vec cells by Western blotting and qRT-PCR. Cell migration, invasion, and sphere formation were examined after Spry2 suppression and overexpression. EGFR-Stat1 and AktERK protein phosphorylation levels were detected via immunoblotting. NEK2 kinase and βcatenin reporter assay were performed for downstream of Spry2 signaling. Results: Although A549-CUG2 cells showed lower levels of the Spry2 protein than A549Vec cells, no difference in levels of Spry2 transcript was observed between both cells via qRT-PCR. Furthermore, MG132 treatment enhanced the protein levels and ubiquitination of Spry2, suggesting that Spry2 protein expression can be regulated via the ubiquitinproteasome pathway. The enforced expression of c-Cbl, known as the binding partner of Spry2, decreased the Spry2 protein levels, whereas its knockdown oppositely increased them. Epithelial–mesenchymal transition (EMT) and sphere formation were increased in A549-Vec cells during Spry2 siRNA treatment, confirming the role of Spry2 in CUG2-induced oncogenesis. Furthermore, EMT and sphere formation were determined by the Spry2 protein levels through the regulation of EGFR-Stat1 and β-catenin-NEK2-Yap1 signaling pathways. Conclusion: CUG2 reduces Spry2 protein levels, the negative signaling molecule of cell proliferation, via c-Cbl, possibly activating the EGFR and β-catenin signaling pathways and, in turn, contributing to the induction of cancer stem cell-like phenotypes.
Citation
Cancer Management and Research, 12, 10243-10250, 2020