Bioinformatics analysis and experimental verification of NLRX1 as a prognostic factor for esophageal squamous cell carcinoma

Abstract

Abstract Background NOD-like receptor X1 (NLRX1), a member of the nucleotide binding and oligomeric domain (NOD) like receptor (NLR) family, is connected to the physiological and pathological processes of inflammation, autophagy, immunity, metabolism, and mitochondrial regulation. It has been discovered to have pro- or anti-tumor effects in various tumor types. However, the biological function of NLRX1 in esophageal squamous cell carcinoma (ESCC) is still unknown. Methods Using bioinformatics methods, the differential expression of NLRX1 at the mRNA level was examined. OS analysis, clinical correlation analysis, ROC analysis, Cox analysis, coexpression analysis, enrichment analysis, immune infiltration analysis, and drug sensitivity analysis were carried out. A nomogram and calibration curve were constructed. Investigating changes in protein expression levels using IHC and WB, assessing the impact on proliferation using CCK-8 assays, examining migration using scratch healing assays, examining migration and invasion using transwell assays, and analyzing apoptosis using fluorescence and flow cytometry. Results Compared to normal tissue adjacent to cancer, NLRX1 is low expressed in ESCC, and patients with low NLRX1 expression have a shorter survival period. NLRX1 is an independent prognostic factor for ESCC and is connected to tumor grading. The low NLRX1 group showed a decrease in NK cells activated, Monocytes, and Macrophages M0 infiltration, and these immune cell infiltration levels were positively correlated with NLRX1 expression. Knocking down NLRX1 promotes the growth and development of KYSE450 cells, while overexpression of NLRX1 inhibits the growth and development of ECA109 cells. NLRX1 negatively regulates the PI3K/AKT pathway in ESCC. Conclusions Through several mechanisms, NLRX1 suppresses tumor growth in ESCC. This offers a fresh viewpoint for investigating the causes and progression of ESCC as well as for searching for more potent therapeutic approaches.