IMUP promotes fatty acid metabolism and lung cancer progression by regulating TGF-β/SMAD3 signaling pathway

Purpose: The purpose of this study was to investigate the biological role of immortalization-upregulated protein (IMUP) in lung Cancer.

Methods: Through bioinformatics analysis, IMUP expression in lung Cancer tissues, overall survival (OS) of patients with lung Cancer, and the biological pathways related to IMUP were analyzed. IMUP expression was assessed utilizing immunohistochemistry staining, RT-qPCR, and western blot. Lung Cancer cells' biological behaviors were determined through flow cytometry, CCK-8, EdU, wound healing, and transwell assays. IMUP's influences on lipid content were assessed using BODIPY 493/503 staining. Western blot was utilized for checking the changes of epithelial-mesenchymal transition (EMT) related proteins (E-cadherin, N-Cadherin, Slug, and Vimentin), some key lipid metabolism proteins (FASN, ACC, and ACSS2) and proteins related to TGF-β signaling pathway (TGF-β, SMAD3, and p-SMAD3). The mouse xenograft model was established to evaluate IMUP's role in lung Cancer growth in vivo.

Results: In lung Cancer tissues and cells, the upregulated IMUP was observed. Patients with high IMUP expression displayed a poorer OS. IMUP knockdown inhibited lung Cancer cell proliferation, viability, migration, invasion, and EMT, but induced cell Apoptosis. Lower lipid content and the levels of FASN, ACC, and ACSS2 were observed in lung Cancer cells with downregulated IMUP. However, the above malignant biological behaviors displayed the contrary results in lung Cancer cells with upregulated IMUP. Notably, IMUP overexpression increased TGF-β and p-SMAD3/SMAD3 expression. On the contrary, IMUP knockdown inhibited TGF-β and p-SMAD3/SMAD3 expression. Besides that, the suppression of IMUP knockdown on lung Cancer cell proliferation, invasion, and fatty acid metabolism was reversed by TGF-β, the promotive effects of IMUP overexpression on cell proliferation, invasion, and fatty acid metabolism were reversed by SIS3 (a SMAD3 inhibitor). In vivo, IMUP knockdown suppresses lung Cancer growth, increased E-cadherin expression, and decreased the expression of Ki67, N-Cadherin, FASN, ACC, and TGF-β.

Conclusion: IMUP knockdown inhibited TGF-β/SMAD3 signaling pathway, thereby restraining fatty acid metabolism and lung Cancer progression.

Cell invasion; Fatty acid metabolism; IMUP; Lung cancer; TGF-β/SMAD3 signaling pathway.