Identification of a Metastasis-Related Protein IFI16 in Esophageal Cancer using a Proteomic Approach

Abstract

Background: Metastasis is the leading cause of the high morality of esophageal squamous cell carcinoma (ESCC), so early monitoring metastasis of esophageal cancer is the key to improve the survival rate of ESCC patients. However, there have not been effective biomarkers for predicting metastasis of ESCC patients,it is an urgent need to identify ESCC metastasis-related proteins. Methods: iTRAQ-based proteomic method was performed in highly metastatic 30M cell established in our previous study and the corresponding parental cells KYSE30.The expression of IFI16 was verified using western blotting and immunohistochemistry (IHC). Then, cck8, transwell assay,mouse metastasis experiments were performed to determine the functional role of IFI16 in esophageal cancer. Finally, RAN-Seq, qpcr, transwell assay were used to investigate the underlying mechanism of IFI16 in esophageal cancer metastasis. Results: The data showed that IFI16 was upregulated in 30M cell compared with KYSE30 cell. IFI16 also increased in ESCC tumor compared with non-tumor tissue. Kaplan-Meier survival curve analysis showed that the relapse-free survival (RFS) of patients with high IFI16 level was worse than that of patients with low IFI16 level (P=0.0449). In addition, IFI16 knockdown did not affect the cell growth, but inhibited ESCC cell migration and invasion in ESCC cells. Moreover, IFI16 knockdown suppressed the lung metastasis of 30M cells in mouse models. Finally, we performed an RNA-Seq assay in IFI16-knocking down 30M cells and identified that knocking down IFI16 downregulated the expression of fibroblast growth factor proteins (FGF1, FGF2 etc.). Furthermore, overexpressing FGF1 and FGF2 rescued the lost of migration and invasion ability of 30M mediated by IFI16 knockdown. Conclusion: Our results demonstrated that IFI16 was a key ESCC metastasis-related protein and played a role in ESCC metastasis through promoting the FGF proteins expression.

IFI16 Induced by Direct Interaction between Esophageal Squamous Cell Carcinomas and Macrophages Promotes Tumor Progression via Secretion of IL-1α

“…In this study, our cDNA microarray analysis revealed that IFI16 expression is upregulated in ESCC cell lines when co-cultured with macrophages. This finding aligns with research by Wang et al, who identified high IFI16 expression in highly metastatic ESCC cells using a proteomics approach [26]. They characterized IFI16 as a protein associated with metastasis in ESCC and observed that silencing IFI16 led to decreased levels of fibroblast growth factor proteins FGF1 and FGF2 in 30M cells, a derivative of the KYSE30 cell line and a model for ESCC metastasis.…”

“…Following the report of FGF proteins [26], since cytokines whose expression were regulated by IFI16 may be involved in the malignant phenotype of ESCC, we also searched comprehensively for cytokines whose expression is downregulated by IFI16 knockdown using a cytokine array. This cytokine array also included FGF2 (alternative name FGF basic, C9 and C10 spots in Supplementary Figure S1), but we could not detect any FGF2 spots in our experiments.…”

“…The results demonstrated the suppression of survival, growth, and migration in the IFI16-silenced ESCC cell lines compared to the siNC-transfected ESCC cell lines (Figures 2C-E and S3A). IFI16 is known to be a positive regulator of cytokine secretion [26,33,34]. Therefore, we conducted a cytokine array to identify humoral factors regulated by IFI16 in TE-11 cells transfected with siIFI16 or siNC (Figure S1A,B).…”

“…The human HIN-200 (hematopoietic, interferon-inducible nuclear proteins with a 200 amino acid repeat) family is a group of interferon-induced genes, with each protein possessing either one or two 200 amino-acid sequence domains at the C-terminus that mediate protein-protein interactions [25]. This group includes interferon-inducible protein 16 (IFI16), myeloid cell nuclear differentiation antigen (MNDA), absent in melanoma 2 (AIM2), and pyrin and HIN domain family member 1 (PYHIN1), each of which has been reported to be related to tumor progression [26][27][28][29]. In this study, we focused on the role of the HIN200 family in the malignant phenotype enhancement of ESCC cells through direct co-culture with macrophages.…”

Changes in intracellular and extracellular proteins after ERGIC3 knockdown in lung cancer: Proteins interacting with ERGIC3, HORN, and FLNA

“…Quantitative proteomics has been widely used in oncology to identify biomarkers with diagnostic and therapeutic potential (Wang et al, 2018;Wang et al, 2022). Isobaric tags for relative and absolute quantitation (iTRAQ), a new protein quantitation technique, has the advantages of a simpler labeling process, higher sensitivity, and a wider application range compared with traditional quantitative methods.…”