Facilitate Tumor Migration Research Articles

Fibrinogen and tumors.

Elevated plasma fibrinogen (Fg) levels consistently correlate with an unfavorable prognosis in various tumor patient cohorts. Within the tumor microenvironment, aberrant deposition and expression of Fg have been consistently observed, interacting with multiple cellular receptors and thereby accentuating its role as a regulator of inflammatory processes. Specifically, Fg serves to stimulate and recruit immune cells and pro-inflammatory cytokines, thereby contributing to the promotion of tumor progression. Additionally, Fg and its fragments exhibit dichotomous effects on tumor angiogenesis. Notably, Fg also facilitates tumor migration through both platelet-dependent and platelet-independent mechanisms. Recent studies have illuminated several tumor-related signaling pathways influenced by Fg. This review provides a comprehensive summary of the intricate involvement of Fg in tumor biology, elucidating its multifaceted role and the underlying mechanisms.

Identification of TBK1 inhibitors against breast cancer using a computational approach supported by machine learning.

Introduction: The cytosolic Ser/Thr kinase TBK1 is of utmost importance in facilitating signals that facilitate tumor migration and growth. TBK1-related signaling plays important role in tumor progression, and there is need to work on new methods and workflows to identify new molecules for potential treatments for TBK1-affecting oncologies such as breast cancer. Methods: Here, we propose the machine learning assisted computational drug discovery approach to identify TBK1 inhibitors. Through our computational ML-integrated approach, we identified four novel inhibitors that could be used as new hit molecules for TBK1 inhibition. Results and Discussion: All these four molecules displayed solvent based free energy values of -48.78, -47.56, -46.78 and -45.47Kcal/mol and glide docking score of -10.4, -9.84, -10.03, -10.06Kcal/mol respectively. The molecules displayed highly stable RMSD plots, hydrogen bond patterns and MMPBSA score close to or higher than BX795 molecule. In future, all these compounds can be further refined or validated by in vitro as well as in vivo activity. Also, we have found two novel groups that have the potential to be utilized in a fragment-based design strategy for the discovery and development of novel inhibitors targeting TBK1. Our method for identifying small molecule inhibitors can be used to make fundamental advances in drug design methods for the TBK1 protein which will further help to reduce breast cancer incidence.

LILRB4 promotes tumor metastasis by regulating MDSCs and inhibiting miR-1 family miRNAs

ABSTRACT Myeloid-derived suppressor cells (MDSCs) are a population of immune suppressive cells that are involved in tumor-associated immunosuppression, and dominate tumor progression and metastasis. In this study, we report that the leukocyte immunoglobulin-like receptor subfamily B member 4 (LILRB4, murine ortholog gp49B) orchestrates the polarization of MDSCs to exhibit pro-tumor phenotypes. We found that gp49B deficiency inhibited tumor metastases of cancer cells, and reduced tumor-infiltration of monocytic MDSCs (M-MDSCs) in tumor-bearing mice. Gp49B−/− MDSCs inhibited pro-tumor immune responses, such as activation of Treg cells, promotion of cancer cell migration, and stimulation of tumor angiogenesis. Treatment of wild-type tumor-bearing mice with gp49B−/− M-MDSCs reduced cancer metastasis. Furthermore, gp49B knockout affected plasma exosome composition in terms of increased miR-1 family microRNAs (miRNAs) expression, which correlates with the upregulation of gp49B−/− MDSC-derived anti-tumor miRNAs. Collectively, our findings reveal that LILRB4/gp49B promotes MDSC-mediated tumor metastasis by regulating the M2-polarization of MDSCs and suppressing the secretion of miR-1 family miRNAs, which facilitate tumor migration and invasion. Abbreviations CTLA-4: cytotoxic T-lymphocyte-associated protein-4; FBS: fetal bovine serum; G-MDSCs: granulocytic-MDSCs; GP49B: glycoprotein 49B; HE: hematoxylin-eosin; ICI: immune checkpoint inhibitor; ITIM: immunoreceptor tyrosine-based inhibition motif; LILRB4: leukocyte immunoglobulin-like receptor B4; M-CSF: macrophage colony stimulating factor; MDSC: myeloid-derived suppressor cell; M-MDSC: monocytic MDSC; MMP-9: metallopeptidase-9; mAb: monoclonal antibody; PBS: phosphate-buffered saline; PCR: polymerase chain reaction; PD-1: programmed death-1; PD-L1: programmed death ligand-1; PMN-MDSC: polymorphonuclear-MDSC; qRT-PCR: quantitative reverse transcription PCR; TAM: tumor associated macrophage; TME: tumor microenvironment; TMM: trimmed mean of M value; VEGFA: vascular endothelial growth factor A

Lasp2 enhances tumor invasion via facilitating phosphorylation of FAK and predicts poor overall survival of non-small cell lung cancer patients.

Lasp2, as well as Lasp1, is a member of the LIM-protein subfamily of the nebulin group characterized by the combined presence of LIM and SH3 domains. Lasp1 and Lasp2 are highly conserved in their LIM, nebulin-like, and SH3 domains but differ significantly at their linker regions. Lasp1 had been described as an oncogenic protein that was highly expressed in diverse cancer types and facilitated tumor proliferation, invasion, and metastasis process. However, unlike Lasp1, little is known about the functions of Lasp2. In the present study, using immunohistochemistry, we found that Lasp2 expression was significantly correlated with histological type (P = 0.012), advanced TNM stage (P = 0.024), positive regional lymph node metastasis (P = 0.035), and poor overall survival (P = 0.001). Would healing assay and transwell assay results indicated that Lasp2 promoted tumor migration and invasion in NSCLC cells. Furthermore, Lasp2 facilitated Snail expression and inhibited Zo-1. The levels of phosphorylated FAK (Tyr397 and Tyr925) were obviously increased after overexpressing Lasp2 and were downregulated by transfecting Lasp2-siRNA. FAK inhibitor counteracted upregulating Snail expression and downregulating of Zo-1 expression induced by Lasp2 overexpression. Taken together, Lasp2 may facilitate tumor migration and invasion of NSCLCs through FAK-Snail/Zo-1 signaling pathway. Lasp2 may be a potential prognostic predictor of NSCLC patients.

Abstract 4449: TET1 inhibits gastric cancer metastasis by demethylation and re-expression of PTEN

Abstract TET1 is a member of ten eleven translocation enzymes, which can convert 5-methylcytosine (5-mC) to 5-hydroxymethylcytosine (5-hmC), eventually to promote CpG islands demethylation of specific gene.. In this study, we found that TET1 expression and 5-hmC content were low in tumor tissues compared to its adjacent non-tumor tissues of gastric cancer. Cell proliferation, migration and invasion were enhanced when TET1 knocked down in gastric cancer cell in vitro and in vivo, while it was suppressed when TET1 over-expressed. We also found that PTENis one of the target genes of TET1. TET1 directly binds to the promoter region of PTEN, activates its transcription through demethylating its CpG islands. Knockdown of TET1 results in hypermethylation of PTEN promoter and low expression of PTEN. PTEN knockdown activates AKT and FAK pathway, which are suppressed by PTEN. The activation of PTEN downstream AKT and FAK facilitates tumor migration, invasion and accelerates cell growth. In conclution, we found a novel mechanism that TET1 suppresses tumor cell growth, migration and invasion through conversion PTEN promoter CpG island methylation to demehylation and then re-expression PTEN. Citation Format: Bingya Liu, Yao-fei Pei, Jian-fang Li, Liping Su, Ran Tao, Min Yan, Zhang-gang Zhu. TET1 inhibits gastric cancer metastasis by demethylation and re-expression of PTEN. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4449.

Klotho Endows Hepatoma Cells with Resistance to Anoikis via VEGFR2/PAK1 Activation in Hepatocellular Carcinoma

Klotho was originally characterized as an aging suppressor gene that predisposed Klotho-deficient mice to premature aging-like syndrome. Although Klotho was recently reported to exhibit tumor suppressive properties during various malignant transformations, the functional role and molecular mechanism of Klotho in hepatocarcinogenesis remains poorly understood. In our present study, immunohistochemical Klotho staining levels in a clinical follow-up of 52 hepatoma patients were significantly associated with liver cirrhosis, tumor multiplicity and venous invasion. The overall survival rate of hepatoma patients with high Klotho expression was significantly lower than those patients with low Klotho expression. Moreover, Klotho overexpression increased cellular migration, anchorage-independent growth, and anoikis resistance in hepatoma cells. Klotho overexpression elevated p21-activated kinase 1 (PAK1) expression and shRNA-mediated PAK1 knockdown and kinase activity inhibition with kinase dead mutant PAK1 K299R coexpression or allosteric inhibitor IPA3 treatment reversed anoikis resistance in Klotho-overexpressed hepatoma cells. More importantly, the pivotal significance of upregulated VEGFR2 protein levels mediated by Klotho expression was confirmed by VEGFR2 inhibitor Axitinib and blocking antibody treatment in hepatoma cells. Axitinib treatment sensitized anoikis was reversed by constitutive active mutant PAK1 T423E coexpression in Klotho-overexpressed hepatoma cells. Conversely, knockdown of Klotho reduced VEGFR2/PAK1 dependent anoikis resistance, which could be reversed by PAK1 T423E. These results revealed a novel oncogenic function of Klotho in promoting anoikis resistance via activating VEGFR2/PAK1 signaling, thus facilitating tumor migration and invasion during hepatoma progression, which could provide a putative molecular mechanism for tumor metastasis.

Abstract P1-05-09: FH535 inhibited migration and growth of breast cancer cells

Abstract Background: Given the lack of effective targeted therapies for triple-negative (TN) breast cancer patients, a better understanding of the mechanisms of growth and invasion of these tumors will provide valuable insight into developing novel approaches to lower the mortality associated with TN breast cancer. There is substantial evidence indicating that the canonical Wnt signaling pathway is activated in TN breast cancer tissues. Previous studies report that FH535, a small molecule inhibitor of the Wnt signaling pathway, decreases growth of cancer cells but not normal fibroblast cells, suggesting this pathway plays a role in tumor progression and metastasis. In this study, we test FH535 as a potential inhibitor of migration, invasion, and growth against malignant phenotypes of breast cancer. Methods and Results: We show that FH535 significantly inhibits growth, migration, and invasion of TN breast cancer cells (MDA-MB-231 and HCC38) using established protocols, implicating the Wnt signaling pathway plays a role in tumor progression and metastasis. FH535 is a potent growth inhibitor for TN breast cancer cells (MDA-MB-231 and HCC38), but not in other breast cancer cells (MCF-7, T-47D, or SK-BR-3) when cultured in three dimensional type I collagen gel. Additionally, western blotting analysis shows that treatment of MDA-MB-231 cells with FH535 markedly inhibits the expression of NEDD9, but not FAK, Src, or p130Cas, suggesting specificity of the Wnt signaling pathway in regulating gene expressions. Co-immunoprecipitation studies suggest that NEDD9 is specifically immunoprecipitated with CSPG4, but not with β1 integrin or CD44, implying CSPG4 and NEDD9 form a molecular complex for facilitating tumor migration, invasion, and growth. Statistical analysis (ANOVA) of The Cancer Genome Atlas (TCGA) project gene expression data (https://tcga-data.nci.nih.gov/) suggests that tumors from patients with basal-type breast cancer had significantly higher CSPG4 expression than tumors from patients with other subtypes, with all the p values <0.0001. Moreover, Luminal A tumors also had a significantly higher CSPG4 expression than Luminal B tumors (p = 0.0006). Discussion: In this study, we demonstrate the pivotal role of the canonical Wnt signaling pathway in enhancing tumor migration, invasion, and growth of TN breast cancer cells MDA-MB-231 and HCC38. We identified NEDD9 as a potential downstream target of the Wnt signaling pathway and found an association between NEDD9 and CSPG4 in MDA-MB-231 cells. Our results suggest a novel mechanism of migration, invasion, and growth of tumor cells involving a molecular complex between CSPG4 and NEDD9, which could be drug targets in TN and possibly Luminal A breast cancer patients. More comprehensive characterization of canonical Wnt signaling pathway by genomic and biological studies of breast cancer cells are clearly required for generating better treatments. The views expressed in this article are those of the author and do not reflect the official policy of the Department of Defense, or U.S. Government. Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P1-05-09.

Abstract 4617: Hu177 cryptic epitope: A novel biomarker for the monitoring of treatment of ovarian cancer

Abstract Background: Distinct cryptic collagen epitopes are among the protein fragments exposed by collagen type IV remodeling, and recent data indicate that these cryptic epitopes may facilitate tumor migration and angiogenesis. In studies with melanoma patients we tested the hypothesis that melanoma can induce detectable changes in systemic levels of cryptic epitope shedding, specifically the HU177 epitope and were able correlate the levels of HU177 shedding with clinical and pathologic parameters (Ng et al Clin Can Res, 14:6253,2008). In addition in a follow-up study the increased shedding of the Hu177 epitope was shown to correlate with a worse prognosis in primary melanoma (Hamilton et al, J Trans Med8:19,2010). In this study we examined wether the serum levels of these unique cryptic epitopes may be useful to monitor, in a phase II study with ovarian patients, the course of the combination treatment of continuous infusion topotecan with erlotinib over multiple cycles of therapeutic intervention. Methods: We made use of our previously published ELISA serum assay (Ng et al) of which the key components involve the anchoring of the primary Hu177 directed antibody followed by blocking with albumin and then incubation with patient serum. The assay is further developed with a sandwich ELISA consisting of biotinylated anti-collagen IV and an anti-biotin antibody conjugated to horseradish peroxidase, followed by chromagenic color development. A parallel examination of the Hu177 epitope was also conducted with a humanized version of the Hu177 antibody (D93). Serial blood samples were monitored for weekly time periods ranging from 6-20 weeks from 5 patients enrolled in this study. Serum were collected and stored in multiple aliquots at –20oC. Results: All 5 patients demonstrated an eventual lowering of shed epitopes, with the time course for a sustained reduction varying with each patient. Once a nadir in the Hu177 epitope occurred it would last from 4-6 weeks. In one patient (KL) the nadir levels were the most sustained and developed late in the course of treatment who had remarkable decreased changes in CA125 that was previously rapidly increasing. This patient had platinum resistant peritoneal carcinomatosis without any dominant masses. Upon failure of topotecan erlotinib as demonstrated by rising markers and development of ascites and a pleural effusion, the shed epitopes rapidly recurred starting at week 18 of the treatment. Comparable serial data were found with the humanized D93 antibody although the sensitivity was more pronounced with the Hu177 antibody. Conclusions: Our interpretation is that this Hu177 determination may provide an indication of active invasion of extracellular matrix (such as in the peritoneum) and may be a useful indicator of a biological effect. We cannot exclude at this point that erlotinib may be contributing to such effects by changes in the matrix. Supported by a grant from the Chemotherapy Foundation Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4617. doi:1538-7445.AM2012-4617

Expression of collagenase-1 (MMP-1) promotes melanoma growth through the generation of active transforming growth factor-β

Tumor cell invasion through basement membranes and into stromal tissue are key steps for promoting growth and metastasis. Tumor cells express various extracellular-matrix-degrading enzymes such as matrix metalloproteinases (MMPs) to degrade extracellular matrix components to facilitate tumor migration and invasion. Histological and clinical studies suggest a role for MMP-1 (collagenase-1) in malignant melanoma invasion. In this study, we evaluated MMP-1 in regulating malignant phenotypes of human melanoma cells by generating human melanoma cells stably transfected with pro-MMP-1 cDNA. The transfectants expressed the active form of MMP-1 associated with cells and showed enhanced invasive and growth abilities in type I collagen gel. Furthermore, MMP-1 expression promoted anchorage-independent growth, which was inhibited in the presence of type II transforming growth factor (TGF)-beta receptor:Fc fusion protein that scavenges TGF-beta receptors. Finally, we demonstrated that MMP-1 directly generated active TGF-beta from its latent form. Thus, these results suggest that MMP-1 produced from melanoma cells would play a role in tumor progression by both degrading matrix proteins and generating active growth factors such as TGF-beta in vivo.