CIRP promotes the progression of non-small cell lung cancer through activation of Wnt/β-catenin signaling via CTNNB1

Abstract

Background Cold-inducible RNA binding protein (CIRP) is a newly discovered proto-oncogene. In this study, we investigated the role of CIRP in the progression of non-small cell lung cancer (NSCLC) using patient tissue samples, cultured cell lines and animal lung cancer models. Methods Tissue arrays, IHC and HE staining, immunoblotting, and qRT-PCR were used to detect the indicated gene expression; plasmid and siRNA transfections as well as viral infection were used to manipulate gene expression; cell proliferation assay, cell cycle analysis, cell migration and invasion analysis, soft agar colony formation assay, tail intravenous injection and subcutaneous inoculation of animal models were performed to study the role of CIRP in NSCLC cells; Gene expression microarray was used to select the underlying pathways; and RNA immunoprecipitation assay, biotin pull-down assay, immunopurification assay, mRNA decay analyses and luciferase reporter assay were performed to elucidate the mechanisms. The log-rank (Mantel-Cox) test, independent sample T-test, nonparametric Mann-Whitney test, Spearman rank test and two-tailed independent sample T-test were used accordingly in our study. Results Our data showed that CIRP was highly expressed in NSCLC tissue, and its level was negatively correlated with the prognosis of NSCLC patients. By manipulating CIRP expression in A549, H460, H1299, and H1650 cell lines, we demonstrated that CIRP overexpression promoted the transition of G1/G0 phase to S phase and the formation of an enhanced malignant phenotype of NSCLC, reflected by increased proliferation, enhanced invasion/metastasis and greater tumorigenic capabilities both in vitro and in vivo. Transcriptome sequencing further demonstrated that CIRP acted on the cell cycle, DNA replication and Wnt signaling pathway to exert its pro-oncogenic action. Mechanistically, CIRP directly bound to the 3′- and 5′-UTRs of CTNNB1 mRNA, leading to enhanced stability and translation of CTNNB1 mRNA and promoting IRES-mediated protein synthesis, respectively. Eventually, the increased CTNNB1 protein levels mediated excessive activation of the Wnt/β-catenin signaling pathway and its downstream targets C-myc, COX-2, CCND1, MMP7, VEGFA and CD44. Conclusion Our results support CIRP as a candidate oncogene in NSCLC and a potential target for NSCLC therapy.

Dihydroartemisinin Reduces Irradiation-Induced Mitophagy and Radioresistance in Lung Cancer A549 Cells via CIRBP Inhibition

“…As a tumor suppressor, CIRBP decreases the proliferation in rectal carcinoma cells and ovarian cancer cells by binds the 5′ and 3′-UTRs of mRNAs, as well as poly U sequences at the 3′-ends to regulate the polyadenylation and 3′-end cleavage, and increases the translational efficiency of DNA damage response genes ATR, Trx-1, and RPA2 [ 28 ]. As a tumor promoter, CIRBP can promote proliferation and reduce apoptosis in breast cancer [ 29 ], prostate cancer [ 30 ], and lung carcinoma cells (A549) [ 31 ] by target mRNA. Importantly, data show that CIRBP induces ROS and mitochondrial DNA fragmentation and regulates macrophage autophagy through TLR4 signaling [ 32 ].…”

Extracellular CIRP Upregulates Proinflammatory Cytokine Expression via the NF-kappaB and ERK1/2 Signaling Pathways in Psoriatic Keratinocytes

“…In addition to its intracellular function as an RNA-binding protein to regulate gene expression [27][28][29], multiple studies have reported that CIRP can be secreted extracellularly and that eCIRP plays important roles in acute and chronic inflammatory diseases. A recent study showed that serum eCIRP levels were upregulated in patients with psoriasis [30], which is consistent with our results.…”

Epidermal growth factor receptor compound and concomitant mutations: advances in precision treatment strategies

“…Patients with TP53 and RB1 mutations have an increased risk of small cell lung cancer [34–36] . Mutations in PIK3CA and PTEN could activate the phosphatidylinositol 3 kinase (PI3K)/protein kinase B (AKT) pathway, and the CTNNB1 mutation could affect the Wnt/β-catenin pathway and result in tumor proliferation and progression [37–40] . Amplification of CDK4 , CDK6 , and CCNE1 affects the transition of the G1/S cell cycle [41] .…”