Coflin‑1, LIMK1 and SSH1 are diferentially expressed in locally advanced colorectal cancer and according to consensus molecular subtypes

Abstract

Background: Colorectal cancer (CRC) is among the deadliest cancers, wherein early dissemination of tumor cells, and consequently, metastasis formation, are the main causes of mortality and poor prognosis. Coflin-1 (CFL-1) and its modulators, LIMK1/SSH1, play key roles in mediating the invasiveness by driving actin cytoskeleton reorganization in various cancer types. However, their clinical signifcance and prognostic value in CRC has not been fully explored. Here, we evaluated the clinical contribution of these actin regulators according to TNM and consensus molecular subtypes (CMSs) classifcation. Methods: CFL-1, LIMK1 and SSH1 mRNA/protein levels were assessed by real-time PCR and immunohistochemical analyses using normal adjacent and tumor tissues obtained from a clinical cohort of CRC patients. The expression levels of these proteins were associated with clinicopathological features by using the chi square test. In addition, using RNA-Seq data of CRC patients from The Cancer Genome Atlas (TCGA) database, we determine how these actin regulators are expressed and distributed according to TNM and CMSs classifcation. Based on gene expression profl‑ ing, Kaplan–Meier survival analysis was used to evaluated overall survival. Results: Bioinformatic analysis revealed that LIMK1 expression was upregulated in all tumor stages. Patients with high levels of LIMK1 demonstrated signifcantly lower overall survival rates and exhibited greater lymph node meta‑ static potential in a clinical cohort. In contrast, CFL-1 and SSH1 have expression downregulated in all tumor stages. However, immunohistochemical analyses showed that patients with high protein levels of CFL-1 and SSH1 exhibited greater lymph node metastatic potential and greater depth of local invasion. In addition, using the CMSs classifca‑ tion to evaluate diferent biological phenotypes of CRC, we observed that LIMK1 and SSH1 genes are upregulated in immune (CMS1) and mesenchymal (CMS4) subtypes. However, patients with high levels of LIMK1 also demonstrated signifcantly lower overall survival rates in canonical (CMS2), and metabolic (CMS3) subtypes. Conclusions: We demonstrated that CFL-1 and its modulators, LIMK1/SSH1, are diferentially expressed and associ‑ ated with lymph node metastasis in CRC. Finally, this expression profle may be useful to predict patients with aggres‑ sive signatures, particularly, the immune and mesenchymal subtypes of CRC.