Thrombospondin-1 Overexpression Induces Immunogenic Cell Death In Mucoepidermoid Carcinoma

To investigate the role and mechanism of thrombospondin-1 (TSP-1) overexpression in inducing immunogenic cell death (ICD) of mucoepidermoid carcinoma (MEC) The MC-3 salivary gland mucoepidermoid carcinoma cell line was selected as the research object, and the experiment was divided into three groups: blank control (Control), overexpressing negative control virus (OE-NC), and overexpressing TSP-1 (OE-TSP-1). Lentiviral vectors were constructed to generate TSP-1 overexpressing stable cell lines, validated by qPCR and Western blot; Proliferation and apoptosis were evaluated via CCK8, colony formation, and apoptosis assays; CRT membrane exposure was analyzed by immunofluorescence, flow cytometry, and membrane protein Western blot; HMGB1 release was detected by ELISA, and ATP secretion was measured using a commercial kit; Phosphorylation levels of the PERK/eIF2α pathway were determined by Western blot. Successful establishment of the TSP-1 overexpressing MC-3 cell line was confirmed by fluorescence microscopy, qPCR, and western blot analysis after lentiviral transfection and selection; The OE-TSP-1 group showed significantly reduced proliferation, colony formation, and increased apoptosis (P<0.05); TSP-1 overexpression significantly enhanced CRT membrane exposure (P<0.05); Both HMGB1 release and ATP secretion were significantly upregulated in the OE-TSP-1 group(P<0.05); The expression of p-PERK and p-eIF2α was significantly increased in the OE-TSP-1 group (P<0.05) TSP-1 overexpression induces ICD in MEC cells through activation of the PERK/eIF2α pathway.