The ability of LCRMP-1 to promote cancer invasion by enhancing filopodia formation is antagonized by CRMP-1

Abstract

Metastasis is a predominant cause of death in patients with cancer. It is a complex multistep process that needs to be better understood if we are to develop new approaches to managing tumor metastasis. Tumor cell invasion of the local stroma is suppressed by collapsin response mediator protein-1 (CRMP-1). Recently, we identified a long isoform of CRMP-1 (LCRMP-1), expression of which correlates with cancer cell invasiveness and poor clinical outcome in patients with non-small-cell lung cancer (NSCLC).Here, we report that LCRMP-1 overexpression in noninvasive human cell lines enhanced filopodia formation, cancer cell migration, and invasion via stabilization of actin. This effect required a highly conserved N-terminal region of LCRMP-1 as well as the WASP family verprolin-homologous protein-1/actin nucleation pathway (WAVE-1/actin nucleation pathway). Furthermore, LCRMP-1 appeared to act downstream of Cdc42, a Rho family protein known to be involved in actin rearrangement. In addition, LCRMP-1 associated with CRMP-1, which downregulated cancer cell metastasis by interrupting the association of LCRMP-1 and WAVE-1. Finally, we found that high-level expression of LCRMP-1 and low-level expression of CRMP-1 were associated with lymph node metastasis and poor survival in patients with NSCLC. In sum, we show that LCRMP-1 and CRMP-1 have opposing functions in regulating cancer cell invasion and metastasis and propose that this pathway may serve as a potential anticancer target. IntroductionCancer metastasis, which is the major cause of treatment failure in cancer patients, is a complex process that involves basement membrane degradation, cell migration, stromal (local) invasion, angiogenesis, intravasation into the circulatory system, adhesion, extravasation into the parenchyma of distant tissues, and colonization (1-3). These processes are regulated by numerous metastasis-promoting and -suppressing genes (4). Thus, identifying novel metastatic genes and their action mechanisms may provide new insights into the pathogenesis and management of cancer metastasis.We previously identified collapsin response mediator protein-1 (CRMP-1) as a novel invasion suppressor and showed that CRMP-1 expression is negatively associated with cell invasiveness and positively associated with better clinical outcomes in patients with non-small-cell lung cancer (NSCLC) (5). Recent studies (6, 7) have shown that CRMP-1 is functionally involved in connective tissue growth factor-mediated inhibition of invasion and metastasis in human lung adenocarcinoma.The CRMPs comprise a family of 5 cytosolic phosphoproteins that inhibit extension of the axonal growth cone during neuronal development (8-11). The members of the CRMP family are closely related 60- to 66-kDa proteins that share 50%-70% amino acid sequence homology and are capable of forming heterotetramers (8,(11)(12)(13)(14). These proteins are distributed mainly in the lamellipodia and filopodia of a neuron's axonal growth cone (14, 15), in which they mediate the signaling pathways that contr...

Estrogen receptorβ2 regulates interlukin-12 receptorβ2 expression via p38 mitogen-activated protein kinase signaling and inhibits non-small-cell lung cancer proliferation and invasion

“…In a heated antigen retrieval process (24), the slides were placed in an EDTA buffer (pH 8.0) and heated for 2 min in a steamer. The slides were incubated overnight at 4˚C with monoclonal mouse anti-human anti-ERβ2 (Serotec) and polyclonal goat anti-human anti-IL-12 Rβ2 (Santa Cruz Biotechnology, Inc., Dallas, TX, USA) antibodies (25) in bovine serum albumin prior to incubating with the secondary antibodies rabbit anti-mouse immunoglobulin G (IgG) k-chain specific antibody (SAB3701212; 1:100, Sigma-Aldrich) or rabbit anti-goat IgG F(ab')2 (SAB3700244; 1:100; Sigma-Aldrich) at RT for 20 min. Color was developed in 3,3'-diaminobenzidine (DAB) solution for 10 min followed by counterstaining with Harris hematoxylin.…”

Long form collapsin response mediator protein-1 promotes the migration and invasion of osteosarcoma cells

“…A previous study showed that LCRMP-1 overexpression promoted cancer cell invasion in vitro and in vivo, and that it was associated with poor overall and disease-free survival in NSCLC patients (8). It has also been shown that LCRMP-1 enhances filopodia formation and is phosphoregulated by glycogen synthase kinase 3β in NSCLC cells (9,13). The results of these previous studies led the present study to investigate the role of LCRMP1 in OS progression.…”

“…Therefore, novel approaches for diagnosis and novel drug targets are required. It has been reported that LCRMP-1 promotes the migration and invasion of NSCLC cells (8,9). In addition, increased expression of N-cadherin and MMPs following LCRMP-1 upregulation has confirmed that LCRMP-1 has a significant role in the migration, invasion and metastasis of tumor cells, as N-cadherin and MMPs are involved in epithelial-mesenchymal transition and remodelling of the extracellular matrix (19,20).…”

“…Pan et al identified a novel isoform of CRMP family proteins in 2008, which was termed long form CRMP-1 (LCRMP-1) (8). In contrast to CRMP-1, which inhibits the migration and invasion of NSCLC, LCRMP-1 promotes filopodia formation, cancer cell migration and invasion, resulting in poor clinical outcomes in NSCLC patients (9). Therefore, LCRMP-1 is an invasion enhancer in NSCLC.…”

Insight into the role of CRMP2 (collapsin response mediator protein 2) in T lymphocyte migration

“…24 In fact, CRMP1 can suppress cancer cell invasion through F-actin depolymerization, while its isoform, LCRMP1, can promote cancer cell migration through actin polymerization and filopodia formation. 25 This is an interesting example of a negative feedback mechanism by protein isoforms in a living cell.…”