Annie Cristhine Moraes Sousa-Squiavinato1, Renata Ivo Vasconcelos1, Adriana Sartorio Gehren1, Priscila Valverde Fernandes2, Ivanir Martins de Oliveira2, Mariana Boroni3, Jose Andrés Morgado-Díaz4. 1. Cellular and Molecular Oncobiology Program, Brazilian National Cancer Institute (INCA), 37 André Cavalcanti Street, 3th Floor, Rio de Janeiro, RJ, 20231-050, Brazil. 2. Pathology Division-DIPAT, Brazilian National Cancer Institute (INCA), Rio de Janeiro, Brazil. 3. Bioinformatics and Computational Biology Lab, Division of Experimental and Translational Research, Brazilian National Cancer Institute (INCA), Rio de Janeiro, Brazil. 4. Cellular and Molecular Oncobiology Program, Brazilian National Cancer Institute (INCA), 37 André Cavalcanti Street, 3th Floor, Rio de Janeiro, RJ, 20231-050, Brazil. jmorgado@inca.gov.br.
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. Cofilin-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 significance 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) classification. 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 classification. Based on gene expression profiling, 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 significantly lower overall survival rates and exhibited greater lymph node metastatic 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 classification to evaluate different 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 significantly lower overall survival rates in canonical (CMS2), and metabolic (CMS3) subtypes. CONCLUSIONS: We demonstrated that CFL-1 and its modulators, LIMK1/SSH1, are differentially expressed and associated with lymph node metastasis in CRC. Finally, this expression profile may be useful to predict patients with aggressive signatures, particularly, the immune and mesenchymal subtypes of CRC.
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. Cofilin-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 significance 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) classification. 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 classification. Based on gene expression profiling, 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 significantly lower overall survival rates and exhibited greater lymph node metastatic 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 classification to evaluate different 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 significantly lower overall survival rates in canonical (CMS2), and metabolic (CMS3) subtypes. CONCLUSIONS: We demonstrated that CFL-1 and its modulators, LIMK1/SSH1, are differentially expressed and associated with lymph node metastasis in CRC. Finally, this expression profile may be useful to predict patients with aggressive signatures, particularly, the immune and mesenchymal subtypes of CRC.
Authors: Jing Yang; Parker Antin; Geert Berx; Cédric Blanpain; Thomas Brabletz; Marianne Bronner; Kyra Campbell; Amparo Cano; Jordi Casanova; Gerhard Christofori; Shoukat Dedhar; Rik Derynck; Heide L Ford; Jonas Fuxe; Antonio García de Herreros; Gregory J Goodall; Anna-Katerina Hadjantonakis; Ruby Y J Huang; Chaya Kalcheim; Raghu Kalluri; Yibin Kang; Yeesim Khew-Goodall; Herbert Levine; Jinsong Liu; Gregory D Longmore; Sendurai A Mani; Joan Massagué; Roberto Mayor; David McClay; Keith E Mostov; Donald F Newgreen; M Angela Nieto; Alain Puisieux; Raymond Runyan; Pierre Savagner; Ben Stanger; Marc P Stemmler; Yoshiko Takahashi; Masatoshi Takeichi; Eric Theveneau; Jean Paul Thiery; Erik W Thompson; Robert A Weinberg; Elizabeth D Williams; Jianhua Xing; Binhua P Zhou; Guojun Sheng Journal: Nat Rev Mol Cell Biol Date: 2020-04-16 Impact factor: 94.444