CREB5 promotes invasiveness and metastasis in colorectal cancer by directly activating MET

Abstract

Background: cAMP responsive element binding protein 5 (CREB5) is a transcriptional activator in eukaryotic cells that can regulate gene expression. Previously, we found that CREB5 was involved in the occurrence and development of colorectal cancer (CRC) using bioinformatics analysis. However, the biological roles and underlying regulatory mechanism of CREB5 in CRC remain unclear. Methods: Real-time PCR, western blotting, and immunohistochemistry were used to examine CREB5 expression. In vitro experiments including migration assay, wound-healing assay, chicken chorioallantoic membrane assay, and human umbilical vein endothelial cells tube formation assay were used to investigate the effects of CREB5 on CRC cell migration and tumor angiogenesis ability. Additionally, an orthotopic implantation assay was performed in nude mice to confirm the effects of CREB5 in vivo. Furthermore, gene set enrichment analysis was performed to explore the potential mechanism of CREB5 in CRC. Results: We found that CREB5 expression was highly upregulated in CRC. CREB5 overexpression was positively correlated with advanced WHO stages and TNM stages and shorter survival in CRC patients. Moreover, CREB5 overexpression promoted while CREB5 silencing reduced the invasiveness and metastatic capacity of CRC cells both in vitro and in vivo. Furthermore, CREB5 directly interacted with the MET promoter and activated the hepatocyte growth factor-MET signalling pathway. Importantly, inhibition of MET reduced the invasion and metastasis of CREB5-overexpressing CRC cells, suggesting that CREB5 promotes metastasis mainly through activation of MET signalling. Conclusion: Our study demonstrates a crucial role for CREB5 in CRC metastasis by directly upregulating MET expression. CREB5 may be both a potential prognostic marker and a therapeutic target to effectively overcome metastasis in CRC.

Tumor cell-derived SPON2 promotes M2-polarized tumor-associated macrophage infiltration and cancer progression by activating PYK2 in CRC

“…Western blotting analysis was performed as previously described [ 44 ] using anti-SPON2 (A12077, ABclonal), anti-PYK2 (bs-3357R, Bioss), anti-p-PYK2 (bs-3400R, Bioss), anti-FAK (bs-1340R, Bioss), anti-Zyxin (60,254–1-Ig, Proteintech), anti-RhoA (HPA062346, Sigma-Aldrich), anti-cortactin (11,381–1-AP, Proteintech), anti-IL10 (20,850–1-AP, Proteintech), anti-CCL2 (66,272–1-Ig, Proteintech), and anti-CSF1 (14,779–1-AP, Proteintech) antibodies. Mouse monoclonal anti-α-tubulin antibody (RM2007, RayAntibody) was used as the internal control.…”

Suppression of MiR130a-3p Using CRISPR/Cas9 Induces Proliferation and Migration of Non-Small Cell Lung Cancer Cell Line

“…(40) MET is a transmembrane tyrosine kinase that are frequently reported in various types of cancers including in NSCLC and colorectal cancer. (41,42) Study in lung cancer cell line has shown that the MET was involved in the regulation of cell migration and invasion. (17) On the other hand, ACVR1, or Activin receptor 1, has been identified as one of the target gene that regulate metastasis, angiogenesis and lymph angiogenesis in gastric cancer.…”

FOXS1 Promotes Tumor Progression by Upregulating CXCL8 in Colorectal Cancer

“…Immunohistochemistry (IHC) staining and scoring were performed as previously described ( 21 ) using anti-FOXS1 (Sigma, HPA042475), anti-CD31 (Abcam, Cambridge, MA, USA, ab9498), and anti-CXCL8 (Bioss, bs-0780R) antibodies. The degree of IHC staining was reviewed and scored independently by two observers.…”