Ceramide Synthase 1 Inhibits Brain Metastasis of Non-Small Cell Lung Cancer by Interacting with USP14 and Downregulating the PI3K/AKT/mTOR Signaling Pathway

SIMPLE SUMMARY: Brain metastasis is common in patients with non-small cell lung cancer and is associated with a poor prognosis. Ceramide synthase 1 participates in malignancy development, but its potential role in non-small cell lung cancer brain metastasis remains unclear. Using bioinformatics analysis and molecular biotechnology, we found that ceramide synthase 1 could inhibit non-small cell lung cancer brain metastasis in vivo and in vitro. Mechanistically, ceramide synthase 1 interacted with ubiquitin-specific protease 14 and inhibited brain metastasis progression by downregulating the PI3K/AKT/mTOR signaling pathway. We suggest that ceramide synthase 1 is an effective therapeutic target for non-small cell lung cancer patients with brain metastases. ABSTRACT: Brain metastasis (BM) is common in patients with non-small cell lung cancer (NSCLC) and is associated with a poor prognosis. Ceramide synthase 1 (CERS1) participates in malignancy development, but its potential role in NSCLC BM remains unclear. This study aimed to explore the physiological effects and molecular mechanism of CERS1 in NSCLC BM. CERS1 expression was evaluated in NSCLC tissues and cell lines, and its physiological roles were subsequently explored in vivo and in vitro. Mass spectrometry and co-immunoprecipitation were performed to explore CERS1-interacting proteins. The associated signaling pathways of CERS1 in NSCLC BM were further investigated using bioinformatics analysis and molecular biotechnology. We demonstrated that CERS1 was significantly downregulated in NSCLC cell lines and BM tissues, and its upregulation was associated with better prognoses. In vitro, CERS1 overexpression inhibited cell migration, invasion, and the ability to penetrate the blood-brain barrier. Moreover, CERS1 interacted with ubiquitin-specific protease 14 (USP14) and inhibited BM progression by downregulating the PI3K/AKT/mTOR signaling pathway. Further, CERS1 expression substantially suppressed BM tumor formation in vivo. This study demonstrated that CERS1 plays a suppressor role in NSCLC BM by interacting with USP14 and downregulating the PI3K/AKT/mTOR signaling pathway, thereby serving as a novel therapeutic target for NSCLC BM.