GOLPH3/CKAP4 promotes metastasis and tumorigenicity by enhancing the secretion of exosomal WNT3A in non-small-cell lung cancer

Abstract

Cancer metastasis is the main cause of mortality associated with non-small-cell lung cancer (NSCLC), accounting for up to 70% of deaths among patients. The mechanisms underlying distal metastasis remain largely unknown. Golgi phosphoprotein 3 (GOLPH3) correlates negatively with overall survival in multiple tumors. In this study, we evaluated the function of GOLPH3 in NSCLC distal metastasis. GOLPH3 was expressed at high levels in samples from patients with NSCLC and was positively associated with clinicopathologic characteristics including clinical stage (P < 0.001), T (P = 0.001), N (P = 0.007), and M (P = 0.001) classification. Functionally, Transwell and wound-healing assays suggested that GOLPH3 overexpression enhances NSCLC cell migration and invasion abilities. Tumor-sphere formation and flow cytometry assays demonstrated that GOLPH3 overexpression enhances a stem cell-like phenotype of NSCLC cells. Metastasis models established by tail vein and intracardiac injection confirmed the pro-metastatic function of GOLPH3 in vivo. A subcutaneous tumor formation model confirmed that GOLPH3 overexpression increased the tumorigenicity of NSCLC cells. Mechanistically, gene set enrichment analysis revealed a positive association of GOLPH3 mRNA expression with WNT-activated gene signatures. Luciferase-reporter and nuclear extract assays showed that GOLPH3 overexpression enhances metastasis and tumorigenicity through activation of the WNT/β-catenin pathway. Immunoprecipitation-mass spectrometry and gene ontology analysis demonstrated that GOLPH3 interacts with cytoskeleton-associated protein 4 (CKAP4) in exosome-mediated distal metastasis. We found that GOLPH3 decreased the amount of plasma membrane-localized CKAP4 and increased the amount of exosome-localized CKAP4 to promote the formation of CKAP4-containing exosomes. Furthermore, we demonstrated that CKAP4 binds exosomal WNT3A to enhance its secretion. Therefore, the GOLPH3/CKAP4 axis plays a crucial role in promoting exosomal-WNT3A secretion to enhance and maintain the stem-like phenotype and metastasis in NSCLC, thus indicating the therapeutic potential of GOLPH3 in patients with NSCLC metastasis.

Linking GOLPH3 and Extracellular Vesicles Content—a Potential New Route in Cancer Physiopathology and a Promising Therapeutic Target is in Sight?

“…Similar results were recently obtained by Song et al 62 in the context of Wnt3A. In their work, the authors studied NSCLC, showing that GOLPH3 overexpression enhances the cell migration and invasion abilities of NSCLC cells.…”

IL‐17 induces non‐small cell lung cancer metastasis via GCN5‐dependent SOX4 acetylation enhancing MMP9 gene transcription and expression

“…Many studies have indicated that IL-17 is upregulated in a variety of cancer tissues and attributed to cancer cell proliferation, metastasis, and so on. [40][41][42] Despite it has been reported that NSCLC metastasis is the major reason for patient death, 43 and the elevation of IL-17 can induce NSCLC metastasis through p38 MAPK or nuclear factor-κB/ ZEB1 activation, 13,14 the exact mechanism for IL-17-triggered NSCLC metastasis is still not elucidated.…”

Molecular subtype identification and signature construction based on Golgi apparatus-related genes for better prediction prognosis and immunotherapy response in hepatocellular carcinoma

“…In addition, GOLM1 can control colitis and colon tumorigenesis by regulating the balance of Notch signaling in the intestine ( 10 ). Golgi phosphoprotein 3 (GOLPH3) is involved in cell migration, GA morphology and orientation, and protein glycosylation, and the increased expression of GOLPH3 observed in a variety of tumors enhances cis-transport of GA and leads to increased cytosolic ejection of pro-metastatic factors such as cytokines, growth factors, and Wnt molecules ( 11 , 12 ). In addition, GA can enhance the secretion of immune factors and promote the formation of an immunosuppressive tumor microenvironment that promotes tumor progression and metastasis.…”