Journal: Translational Lung Cancer Research

Article Title: TIPE3 promotes non-small cell lung cancer progression via the protein kinase B/extracellular signal-regulated kinase 1/2-glycogen synthase kinase 3β-β-catenin/Snail axis

doi: 10.21037/tlcr-21-147

Figure Lengend Snippet: TIPE3 promotes NSCLC progression by an AKT/ERK1/2-GSK3β-β-catenin/Snail axis. (A) WB analysis of the AKT/ERK1/2-GSK3β-β-catenin/Snail axis and its regulatory targets in TIPE3 knock-down and TIPE3-overexpressing NSCLC cells. Representative WB images are shown. (B) IF analysis of the AKT/ERK1/2-GSK3β-β-catenin/Snail axis in TIPE3 knock-down and TIPE3-overexpressing NSCLC xenograft tumor tissues. Representative IF images of merge (red, Alexa Fluor 647; green, GFP; and blue, DAPI) (×200) are shown. (C) WB analysis of β-catenin, Snail1, and Slug in TIPE3-overexpressing NSCLC cells after treatment with PI3K or MEK inhibitor and TIPE3 knock-down NSCLC cells after treatment with GSK3β or proteasome inhibitor. Representative WB images are shown. (D) WB analysis of β-catenin, Snail1, and Slug in TIPE3 knock-down NSCLC cells after β-Trcp or STUB1 knockdown. Representative WB images are shown. (E,F) Clinical association of TIPE3 with the AKT/ERK1/2-GSK3β-β-catenin/Snail axis in lung cancer. Representative IHC images of Case 27 (+++, high TIPE3 expression) and Case 34 (-, low TIPE3 expression) lung cancer T tissues (×400) (E) are shown. Percentage of lung cancer T tissue specimens showing high or low TIPE3 expression in relation to the expression level of proteins related to the AKT/ERK1/2-GSK3β-β-catenin/Snail axis (F) is shown. *, P<0.05, Pearson’s χ2 test. (G) A schematic model for the mechanism of action of TIPE3 during NSCLC progression. Upregulation of TIPE3 promotes growth and metastasis in NSCLC cells by activating β-catenin, Snail1, and Slug transcriptional signaling via the AKT/ERK1/2-GSK3β axis. NSCLC, non-small cell lung cancer; WB: western blotting; AKT, protein kinase B; ERK1/2, extracellular signal-regulated kinase 1/2; GSK3β, glycogen synthase kinase 3β; IF, immunofluorescence; GFP, green fluorescent protein; DAPI: 4’,6-diamidino-2-phenylindole; PI3K, phosphatidylinositol 3-kinase; MEK: mitogen-activated protein kinase kinase; IHC, immunohistochemistry; T, lung cancer tumor.

Article Snippet: Rabbit anti-Snail1 (cat. no. 101167-T10) for IF and IHC was supplied by Sino Biological (Wayne, PA, USA), and rabbit anti-Slug (cat. no. GTX128796) for IHC was supplied by GeneTex (Irvine, CA, USA).

Techniques: Expressing, Western Blot, Immunofluorescence, Immunohistochemistry