• Kupffer cells
• hepatic stellate cells
• liver metastasis
• liver microenvironment
• liver sinusoidal endothelial cells
• neutrophilic
• tumor-associated macrophages

• Humans
• Endothelial Cells/pathology
• Liver/pathology
• Liver Neoplasms/pathology
• Kupffer Cells
• Hepatocytes
• Tumor Microenvironment

• S202203 Suqian Science and Technology Support Project Fund
• BE2015666 Major Clinical Special Fund of Jiangsu Province
• SH2022090 Zhenjiang Social Development Fund
• ZD2022052 The Key Project Fund of the Jiangsu Provincial Health Commission

• CRLM
• cells
• tumor microenvironment

• ACKR3
• chemokine
• colorectal cancer
• epigenetics
• metastasis
• microenvironment
• prognosis
• resistance
• signaling pathways
• therapy

We utilized Fas21, a resveratrol analog, to modulate the function of hepatic stellate cells (HSCs) and liver sinusoidal endothelial cells (LSECs) during the angiogenic phase of murine liver metastasis by B16 melanoma and 51b colorectal carcinoma. Preangiogenic micrometastases were treated with Fas21 (1 mg/kg/day) or vehicle during the development of intra-angiogenic tracts. Mice treated with Fas21 showed reduced liver tumor foci in both liver metastasis models. Micrometastases were classified immunohistochemically, as well as according to their position coordinates and connection to local microvasculature. The volume of liver occupied by sinusoidal-type foci, containing infiltrating angiogenic capillaries, decreased by ~50% in Fas21-treated mice compared to vehicle-treated ones in both tumor metastasis models. The volume of portal foci, containing peripheral neoangiogenesis within a discontinuous layer of myofibroblasts, was similar in all experimental groups in both tumor metastasis models, but displayed enhanced necrotic central areas devoid of angiogenesis following Fas21 treatment. As a result, sinusoidal tumors from mice treated with Fas21 showed a 50% reduction in desmin(+)/asma(+) HSCs and CD31(+) vessel density, and a 45% reduction in intrametastatic VEGF mRNA compared with sinusoidal tumors from vehicle-treated mice. Necrotic portal metastases increased 2-4-fold in treated mice. In vitro, Fas21 reduced VEGF secretion by HSCs and 51b cells dose-dependently. Additionally, HSCs migration in response to tumor soluble factors was dose-dependently diminished by Fas21, as was LSEC migration in response to HSCs and tumor soluble factors. Resveratrol analog Fas21 inhibits the proangiogenic response of HSCs and LSECs during the development of murine liver metastasis.

• Angiogenesis
• Cancer
• Hepatic stellate cell
• Liver metastasis
• Liver sinusoidal endothelial cell
• Tumor microenvironment

• angiogenesis
• antioxidant
• apoptosis
• cancer
• melatonin
• metastasis

• Animals
• Apoptosis
• Cell Movement
• Cell Proliferation
• Humans
• Hypoxia/physiopathology
• Melatonin/therapeutic use
• Neoplasms/blood supply
• Neoplasms/drug therapy
• Neoplasms/metabolism
• Neoplasms/pathology
• Neovascularization, Pathologic/drug therapy
• Signal Transduction

• Cancer treatment
• Cancer-associated fibroblasts
• Metastasis-associated fibroblasts
• Metastatic cancer

Hepatocellular carcinoma (HCC) is a highly malignant tumor with a particularly poor prognosis. The tumor microenvironment (TME) is closely associated with tumorigenesis, progression, and treatment. However, the relationship between TME genes and HCC patient prognosis is poorly understood.

In this study, we identified two prognostic subtypes based on the TME using data from The Cancer Genome Atlas and Gene Expression Omnibus. The Microenvironment Cell Populations-counter method was used to evaluate immune cell infiltration in HCC. Differentially expressed genes between molecular subtypes were calculated with the Limma package, and clusterProfiler was used for Gene Ontology and Kyoto Encyclopedia of Genes and Genomes functional enrichment analyses to identify genes related to the independent subtypes. We also integrated mRNA expression data into our bioinformatics analysis.

We identified 4227 TME-associated genes and 640 genes related to the prognosis of HCC. We defined two major subtypes (Clusters 1 and 2) based on the analysis of TME-associated gene expression. Cluster 1 was characterized by increased expression of immune-associated genes and a worse prognosis than Cluster 2.

The identification of these HCC subtypes based on the TME provides further insight into the molecular mechanisms and prediction of HCC prognosis.

• Hepatocellular carcinoma
• gene signature
• immune classification
• immune microenvironment
• molecular mechanism
• prognostic subtype

• CAF
• Colorectal cancer
• Cyclooxygenase-2
• Hepatic stellate cells
• Liver metastasis
• Tumor microenvironment

• BRAF inhibition
• adjuvant
• cancer nutrition
• chemoresistance
• fibroblasts
• melanoma
• tumor microenvironment

• Animals
• Antineoplastic Agents/pharmacology
• Ascorbic Acid/pharmacology
• Cancer-Associated Fibroblasts/drug effects
• Cell Line, Tumor
• Drug Resistance, Neoplasm/drug effects
• Folic Acid/pharmacology
• Humans
• Melanoma/drug therapy
• Melanoma/genetics
• Mice
• Pantothenic Acid/pharmacology
• Plant Extracts/pharmacology
• Protein Kinase Inhibitors/pharmacology
• Proteomics
• Proto-Oncogene Proteins B-raf/antagonists & inhibitors
• Skin Neoplasms/drug therapy
• Skin Neoplasms/genetics
• Vemurafenib/pharmacology
• Vitamin B 12/pharmacology
• Vitamin B 6/pharmacology
• Zinc Sulfate/pharmacology
• Melanoma, Cutaneous Malignant

• 2016.0036 Catalysis S.L

• capsule
• carcinoma-associated fibroblasts
• desmoplasia
• hepatic stellate cells
• histologic growth patterns
• liver
• microenvironment

• Biomarkers
• Biopsy
• Cancer-Associated Fibroblasts/metabolism
• Cancer-Associated Fibroblasts/pathology
• Colorectal Neoplasms/pathology
• Endothelial Cells/metabolism
• Female
• Hepatic Stellate Cells/metabolism
• Humans
• Immunohistochemistry
• Liver/pathology
• Liver Neoplasms/diagnosis
• Liver Neoplasms/secondary
• Lymphocytes, Tumor-Infiltrating/metabolism
• Lymphocytes, Tumor-Infiltrating/pathology
• Male
• Tumor Microenvironment

• Ocoxin
• anti-inflammatory
• antioxidant
• cancer
• chemorresistance
• coadjuvant
• natural product

• cancer
• immunotherapy
• liver metastasis
• targeted therapy
• transforming growth factor beta
• tumor microenvironment

Innate lymphoid cells (ILCs) are the innate counterparts of adaptive immune cells. Emerging data indicate that they are also key players in the progression of multiple tumors. In this review we briefly describe ILCs’ functions in the skin, lungs and liver. Next, we analyze the role of ILCs in primary cutaneous melanoma and in its most frequent and deadly metastases, those in liver and lung. We focus on their dual anti– and pro-tumoral functions, depending on the cross-interactions among them and with the surrounding stromal cells that form the tumor microenvironment (TME) in each organ. Next, we detail the role of extracellular vesicles secreted to the TME by ILCs and melanoma on both cell populations. We conclude that the identification of markers and tools to allow the modulation of individual ILC subsets, in addition to the development of standardized protocols, is essential for addressing the therapeutic modulation of ILCs.

The role of innate lymphoid cells (ILCs) in cancer progression has been uncovered in recent years. ILCs are classified as Type 1, Type 2, and Type 3 ILCs, which are characterized by the transcription factors necessary for their development and the cytokines and chemokines they produce. ILCs are a highly heterogeneous cell population, showing both anti– and protumoral properties and capable of adapting their phenotypes and functions depending on the signals they receive from their surrounding environment. ILCs are considered the innate counterparts of the adaptive immune cells during physiological and pathological processes, including cancer, and as such, ILC subsets reflect different types of T cells. In cancer, each ILC subset plays a crucial role, not only in innate immunity but also as regulators of the tumor microenvironment. ILCs’ interplay with other immune and stromal cells in the metastatic microenvironment further dictates and influences this dichotomy, further strengthening the seed-and-soil theory and supporting the formation of more suitable and organ-specific metastatic environments. Here, we review the present knowledge on the different ILC subsets, focusing on their interplay with components of the tumor environment during the development of primary melanoma as well as on metastatic progression to organs, such as the liver or lung.

• extracellular vesicles
• innate lymphoid cells
• melanoma
• tumor microenvironment

Liver metastasis depends on the collagenous microenvironment generated by hepatic sinusoidal cells (SCs). DDR1 is an atypical collagen receptor linked to tumor progression, but whether SCs express DDR1 and its implication in liver metastasis remain unknown. Freshly isolated hepatic stellate cells (HSCs), Kupffer cells (KCs), and liver sinusoidal endothelial cells (LSECs), that conform the SCs, expressed functional DDR1. HSCs expressed the largest amounts. C26 colon carcinoma secretomes increased DDR1 phosphorylation in HSCs and KCs by collagen I. Inhibition of kinase activity by DDR1-IN-1 or mRNA silencing of DDR1 reduced HSCs secretion of MMP2/9 and chemoattractant and proliferative factors for LSECs and C26 cells. DDR1-IN-1 did not modify MMP2/9 in KCs or LSECs secretomes, but decreased the enhancement of C26 migration and proliferation induced by their secretomes. Gene array showed that DDR1 silencing downregulated HSCs genes for collagens, MMPs, interleukins and chemokines. Silencing of DDR1 before tumor inoculation reduced hepatic C26 metastasis in mice. Silenced livers bore less tumor foci than controls. Metastatic foci in DDR1 silenced mice were smaller and contained an altered stroma with fewer SCs, proliferating cells, collagen and MMPs than foci in control mice. In conclusion, hepatic DDR1 promotes C26 liver metastasis and favors the pro-metastatic response of SCs to the tumor.

Cancer associated fibroblasts (CAF) and the extracellular matrix (ECM) produced by them have been recognized as key players in cancer biology and emerged as important targets for cancer treatment and drug discovery. Apart from their presence in stroma rich tumors, such as biliary, pancreatic and subtypes of hepatocellular cancer (HCC), both CAF and certain ECM components are also present in cancers without an overt intra-tumoral desmoplastic reaction. They support cancer development, growth, metastasis and resistance to chemo- or checkpoint inhibitor therapy by a multitude of mechanisms, including angiogenesis, ECM remodeling and active immunosuppression by secretion of tumor promoting and immune suppressive cytokines, chemokines and growth factors. CAF resemble activated hepatic stellate cells (HSC)/myofibroblasts, expressing α-smooth muscle actin and especially fibroblast activation protein (FAP). Apart from FAP, CAF also upregulate other functional cell surface proteins like platelet-derived growth factor receptor β (PDGFRβ) or the insulin-like growth factor receptor II (IGFRII). Notably, if formulated with adequate size and zeta potential, injected nanoparticles home preferentially to the liver. Several nanoparticular formulations were tested successfully to deliver dugs to activated HSC/myofibroblasts. Thus, surface modified nanocarriers with a cyclic peptide binding to the PDGFRβ or with mannose-6-phosphate binding to the IGFRII, effectively directed drug delivery to activated HSC/CAF in vivo. Even unguided nanohydrogel particles and lipoplexes loaded with siRNA demonstrated a high in vivo uptake and functional siRNA delivery in activated HSC, indicating that liver CAF/HSC are also addressed specifically by well-devised nanocarriers with optimized physicochemical properties. Therefore, CAF have become an attractive target for the development of stroma-based cancer therapies, especially in the liver.

• HCC
• anti-fibrotic
• desmoplastic stroma
• extracellular matrix (ECM)
• immune suppression
• nanoparticle
• platelet-derived growth factor (PDGF)
• siRNA
• therapy
• tumor microenvironment (TME)
• tumor-associated-macrophage (TAM)

• EMT
• FOXM1
• chrysin
• melanoma
• metastasis
• β-catenin

• Animals
• Cell Line, Tumor
• Cell Proliferation
• Female
• Flavonoids/administration & dosage
• Forkhead Box Protein M1/genetics
• Forkhead Box Protein M1/metabolism
• Humans
• Lung Neoplasms/genetics
• Lung Neoplasms/metabolism
• Lung Neoplasms/prevention & control
• Lung Neoplasms/secondary
• Melanoma/drug therapy
• Melanoma/metabolism
• Melanoma/pathology
• Melanoma/physiopathology
• Mice
• Mice, Inbred C57BL
• Neoplasm Invasiveness
• Neoplasm Metastasis
• Signal Transduction
• beta Catenin/genetics
• beta Catenin/metabolism

Cancer cell migration, tumor angiogenesis, and activated hepatic stellate cells (a-HSCs) promote the development of colorectal liver metastases (CLM). Wiskott–Aldrich syndrome protein family verprolin-homologous protein 2 (WAVE2) has been associated with CLM, although the underlying molecular mechanisms remain unclear.

In the current study, we evaluated the relationship between WAVE2 and CLM in 103 CLM patients who underwent liver resection. Immunohistochemistry (IHC) staining was performed to determine the association between WAVE2 protein expression and hepatic micro-metastasis in human CLM tissues. WAVE2 knockout was performed in hepatic stellate cells (HSC) to explore the function and signaling pathways of WAVE2 in colorectal cancer progression.

Significantly higher levels of WAVE2 were detected in portal-associated relative to sinusoid-associated micro-metastasis. A strong correlation was identified between WAVE2 levels and microvessel density (MVD) in hepatic metastasis. Similarly, expression of WAVE2 was closely associated with activation of HSCs. Mechanistically, WAVE2 regulated the progression of human CLM acts by regulating the growth factor β (TGF-β) and Hippo pathways via effector yes-associated protein (YAP1).

Overall, our results demonstrated that WAVE2 participates in CLM tumor microenvironment, and can be a potential latent therapeutic target for CLM.

• WAVE2
• colorectal cancer
• hepatic stellate cells
• liver metastasis

• BRCA1
• BRCA1-associated genome surveillance complex
• colorectal cancer
• liver metastasis
• miRNAs
• proteomics
• tissue microarray
• transcriptomics
• tumor microenvironment

• Biomarkers
• Colorectal cancer
• Exosomal proteins
• Gene expression profiling
• Inter-organ prometastatic communication
• Liver metastasis
• Liver prometastatic reaction
• Melanoma
• Premetastatic niche
• Tumor microenvironment

• Alpha-smooth muscle actin
• Cirrhosis
• Desmoplasia
• Epithelial-mesenchymal transition
• Fibrosis
• Hepatic stellate cell
• Hepatocellular carcinoma
• Metastasis
• Pancreatic adenocarcinoma
• Pancreatic stellate cell
• Sonic hedgehog
• Vitamin A

• Animals
• Hepatic Stellate Cells/pathology
• Hepatocytes/pathology
• Humans
• Kupffer Cells/pathology
• Liver Neoplasms/secondary
• Neoplasm Metastasis/pathology
• Neoplasms/pathology
• Phagocytes/pathology
• Tumor Microenvironment/physiology

• MR/P018920/1 Medical Research Council
• 25607 Cancer Research UK
• C48719/A25607 Cancer Research UK
• Wellcome Trust
• 102521/Z/13/Z Wellcome Trust

• Angiogenesis
• Cancer-associated fibroblast
• Epithelial-mesenchymal transition
• Extracellular matrix
• Fibroblast activation protein
• Jagged-1
• Matrix metalloproteinase
• Myofibroblast
• Platelet-derived growth factor
• Stromal cell-derived factor-1
• Transforming growth factor-β
• Tumor stroma
• Tumor-associated macrophages
• Tumor-infiltrating leukocyte
• Vascular endothelial growth factor

• activated hepatic stellate cells
• angiogenesis
• angiopoietin-1
• hepatic cellular carcinoma

The mechanism underlying tumor spread through air spaces (STAS) has not been well studied. We investigated the role of tumor stromal cells in the pathogenesis of STAS from a pathological perspective and evaluated the prognostic significance of tumor stromal cells and STAS in postoperative patients with lung adenocarcinoma.

We retrospectively analyzed 208 postsurgical patients with stage I–IIIA lung adenocarcinoma. The presence of STAS was evaluated by hematoxylin and eosin staining. The expression of α‐smooth muscle actin (SMA)‐positive cancer‐associated fibroblasts (CAFs) and CD204‐positive tumor‐associated macrophages (TAMs) was analyzed by immunohistochemistry. A logistic regression model was applied to confirm the predictive factors of STAS. Survival analysis was performed to evaluate the effect of α‐SMA‐positive CAFs, CD204‐positive TAMs, and STAS on prognosis. A nomogram was generated to evaluate the prognosis of postoperative patients.

Logistic regression suggested that the expression of α‐SMA‐positive CAFs (P < 0.001) and the number of CD204‐positive TAMs (P < 0.001) were related to the presence of STAS. The multivariate Cox proportional hazards model suggested that STAS (P = 0.004), α‐SMA‐positive CAFs (P < 0.001), and CD204‐positive TAMs (P < 0.001) were independent risk factors for prognosis. Harrell's c‐indexes for overall and recurrence‐free survival prediction based on nomograms were 0.84 (95% confidence interval 0.76–0.91) and 0.82 (95% confidence interval 0.76–0.89), respectively.

The presence of STAS was associated with high expression of α‐SMA and CD204 in lung adenocarcinoma. Nomograms including STAS and stromal cells as variables are recommended as practical models to evaluate the prognosis of lung adenocarcinoma patients.

• Lung cancer
• STAS
• prognosis
• stromal cell
• tumor spread through air spaces

Mammalian target of rapamycin complex 2 (mTORC2) with its pivotal component rapamycin-insensitive companion of mTOR (RICTOR) is the major regulator of AKT phosphorylation and is increasingly implicated in tumor growth and progression. In cutaneous melanoma, an extremely aggressive and highly metastatic disease, RICTOR overexpression is involved in tumor development and invasiveness. Therefore, we investigated the impact of RICTOR inhibition in melanoma cells in vitro and in vivo with special emphasis on hepatic metastasis. Moreover, our study focused on the interaction of tumor cells and hepatic stellate cells (HSC) which play a crucial role in the hepatic microenvironment. In silico analysis revealed increased RICTOR expression in melanoma cells and tissues and indicated higher expression in advanced melanoma stages and metastases. In vitro, transient RICTOR knock-down via siRNA caused a significant reduction of tumor cell motility. Using a syngeneic murine splenic injection model, a significant decrease in liver metastasis burden was detected in vivo. Moreover, stimulation of melanoma cells with conditioned medium (CM) from activated HSC or hepatocyte growth factor (HGF) led to a significant induction of AKT phosphorylation and tumor cell motility. Blocking of RICTOR expression in cancer cells diminished constitutive and HGF-induced AKT phosphorylation as well as cell motility. Interestingly, RICTOR blockade also led to an abrogation of CM-induced effects on AKT phosphorylation and motility in melanoma cells. In conclusion, these results provide first evidence for a critical role of mTORC2/RICTOR in melanoma liver metastasis via cancer cell/HSC interactions.

• carcinoembryonic antigen
• colorectal cancer
• fibroblasts
• fibronectin
• metastasis

• liver metastasis
• liver sinusoidal endothelial cells
• miRNA
• nanomedicine
• therapy

• Cancer associated fibroblasts
• Growth factors
• Hepatocellular carcinoma
• Inflammatory cytokines

• adhesion molecules
• angiogenesis
• immune response
• intercellular adhesion molecule-1
• invasion
• liver metastasis
• tumor microenvironment

Hepatic stellate cells (HSCs) are critical mediators of immunosuppression and the pathogenesis of hepatocellular carcinoma (HCC). Our previous work indicates that HSCs promote HCC progression by enhancing immunosuppressive cell populations including myeloid-derived suppressor cells (MDSCs) and regulatory T cells (Tregs). MDSCs are induced by inflammatory cytokines (e.g., prostaglandins) and are important in immune suppression. However, how HSCs mediate expansion of MDSCs is uncertain. Thus, we studied activated HSCs that could induce MDSCs from bone marrow cells and noted that HSC-induced MDSCs up-regulated immunosuppressive activity via iNOS, Arg-1, and IL-4Rα. After treating cells with a COX-2 inhibitor or an EP4 antagonist, we established that HSC-induced MDSC accumulation was mediated by the COX2-PGE₂-EP4 signaling. Furthermore, in vivo animal studies confirmed that inhibition of HSC-derived PGE₂ could inhibit HSC-induced MDSC accumulation and HCC growth. Thus, our data show that HSCs are required for MDSC accumulation mediated by the COX2-PGE₂-EP4 pathway, and these data are the first to link HSC and MDSC subsets in HCC immune microenvironment and provide a rationale for targeting PGE₂ signaling for HCC therapy.

• cyclooxygenase 2
• hepatic stellate cell
• hepatocellular carcinoma
• myeloid-derived suppressor cells
• prostaglandin 2

• fluke
• inflammation
• mechanosensitive signaling
• primary sclerosing cholangitis (PSC)
• stiffness
• stroma

Hepatocellular carcinoma (HCC) is one of the most common malignancies characterized by insidious onset and poor prognosis. Studies have shown that β adrenergic receptor signaling, especially β2 adrenergic receptor (β2-AR) signaling, regulates multiple cellular processes that contribute to the initiation and progression of cancer, including differentiation, proliferation and apoptosis. β2-AR signaling is also involved in tumor angiogenesis, progression and metastasis in HCC. Therefore, understanding of the role of the β2 adrenergic receptor signaling pathway in HCC progression and metastasis will be of great value in developing therapeutic strategies for this maliganancy. In this paper, we will discuss the role of β2 adrenergic receptor signaling pathway in HCC.

• β2 adrenergic receptors
• Signaling pathway
• Hepatocellular carcinoma

• Liver metastasis
• hepatic niche
• metastatic models
• microphysiological
• tumor microenvironment

• Biomimetics
• Hepatic Stellate Cells/pathology
• Humans
• Kupffer Cells/pathology
• Liver/pathology
• Neoplasm Metastasis/pathology
• Tumor Microenvironment/physiology

• I01 BX003368 BLRD VA
• UH3 TR000496 NCATS NIH HHS
• P30 CA014051 NCI NIH HHS
• I01 BX001017 BLRD VA
• UH3 TR000503 NCATS NIH HHS

• Angiogenesis
• Cell adhesion
• Hepatic metastasis
• Pancreatic cancer
• Remodeling of extracellular matrix
• Tumor-induced immunosuppression

• Humans
• Liver Neoplasms/immunology
• Liver Neoplasms/pathology
• Liver Neoplasms/secondary
• Liver Neoplasms/therapy
• Pancreatic Neoplasms/immunology
• Pancreatic Neoplasms/pathology
• Pancreatic Neoplasms/therapy

Carcinoma-associated fibroblasts (CAFs) are critical in determining tumor invasion and metastasis. However the role of CAFs in the invasion of salivary gland adenoid cystic carcinoma (ACC) is poorly understood. In this study, we isolated primary CAFs from two ACC patients. ACC-derived CAFs expressed typical CAF biomarkers and showed increased migration and invasion activity. Conditioned medium collected from CAFs significantly promoted ACC cell migration and invasion. Co-culture of CAFs with ACC cells in a microfluidic device further revealed that CAFs localized at the invasion front and ACC cells followed the track behind the CAFs. Interfering of both matrix metalloproteinase and CXCL12/CXCR4 pathway inhibited ACC invasion promoted by CAFs. Overall, our study demonstrates that ACC-derived CAFs exhibit the most important defining feature of CAFs by promoting cancer invasion. In addition to secretion of soluble factors, CAFs also lead ACC invasion by creating an invasive track in the ECM.

• Carcinoma, Adenoid Cystic/metabolism
• Carcinoma, Adenoid Cystic/pathology
• Cell Line, Tumor
• Cell Movement
• Chemokine CXCL12/metabolism
• Coculture Techniques
• Female
• Fibroblasts/metabolism
• Fibroblasts/pathology
• Gelatinases/metabolism
• Humans
• Male
• Neoplasm Invasiveness
• Neoplasm Proteins/metabolism
• Receptors, CXCR4/metabolism
• Salivary Gland Neoplasms/metabolism
• Salivary Gland Neoplasms/pathology

Liver metastatic disease is the main cause of death in colorectal cancer (CRC) patients. During metastatic spread of the disease an imbalance in the oxidative stress and inflammation plays a crucial role in tumor progression. In order to improve the efficacy of current therapies, new complementary therapeutic approaches are being analyzed including biologically active compounds with low side effects. The anti-inflammatory and anti-oxidant properties of Ocoxin^® oral solution (OOS) prompt us to analyze its effect on the metastatic development of CRC to the liver. First, in vitro effect of OOS in tumor cell viability and migration was analyzed. Second, in vivo effect of different dosage patterns and concentrations in the development of hepatic metastasis was analyzed by intrasplenic inoculation of C26 colon carcinoma cells in Balb/c mice. Third, the expression of alpha smooth muscle actin, caspase-3 and Ki-67 expression was quantified by immunohistochemistry, then gene expression levels of inflammatory factors were measured by quantitative RT-PCR. According to our results, OOS reduced tumor cell viability and migration in vitro. Moreover, in vivo daily administration of OOS from the 7th day after tumor cell inoculation decreased the total area and size of metastatic foci in the liver. Furthermore, cell proliferation and fibroblast recruitment was decreased in tumor foci while a higher number of apoptotic cells were observed. Finally, RNA levels for the inflammatory mediators COX-2, IFNγ, IL1β, IL6 and TNFα were reduced in total liver. In conclusion, OOS reduced the metastatic development of colorectal cancer to the liver by increasing apoptosis, and decreasing tumor cell proliferation and fibroblast recruitment in the tumor foci, as well as the expression of inflammatory mediators in total liver. These results point out OOS as a potential supplement to be applied as complementary therapy for the treatment of liver metastasis from colorectal cancer.

• Acidic tumor microenvironment
• Glycolysis
• Hepatic stellate cells
• Proton pump inhibitor

• Acids/chemistry
• Animals
• Biomarkers, Tumor/genetics
• Biomarkers, Tumor/metabolism
• Blotting, Western
• Carcinoma, Hepatocellular/metabolism
• Carcinoma, Hepatocellular/mortality
• Carcinoma, Hepatocellular/secondary
• Cell Movement
• Cell Proliferation
• Culture Media, Conditioned/pharmacology
• Enzyme-Linked Immunosorbent Assay
• Female
• Fluorescent Antibody Technique
• Gene Expression Profiling
• Hepatic Stellate Cells/metabolism
• Hepatic Stellate Cells/pathology
• Humans
• Hydrogen-Ion Concentration
• Immunoenzyme Techniques
• Liver Neoplasms/metabolism
• Liver Neoplasms/mortality
• Liver Neoplasms/pathology
• Mice
• Mice, Inbred BALB C
• Mice, Nude
• Myofibroblasts/metabolism
• Myofibroblasts/pathology
• Osteopontin/genetics
• Osteopontin/metabolism
• RNA, Messenger/genetics
• Real-Time Polymerase Chain Reaction
• Reverse Transcriptase Polymerase Chain Reaction
• Tumor Cells, Cultured
• Tumor Microenvironment
• Xenograft Model Antitumor Assays

Metastatic growth by colorectal cancer cells in the liver requires the ability of the cancer cells to interact with the new microenvironment. This interaction results in three histological growth patterns of liver metastases: desmoplastic, pushing, and replacement. In primary colorectal cancer several proteases, involved in the degradation of extracellular matrix components, are up‐regulated. In liver metastases, their expression is growth pattern dependent. Tissue inhibitor of matrix metalloproteinase‐1 (TIMP‐1) is a strong prognostic marker in plasma from colorectal cancer patients, with significant higher levels in patients with metastatic disease. We therefore wanted to determine the expression pattern of TIMP‐1 in primary colorectal cancers and their matching liver metastases. TIMP‐1 mRNA was primarily seen in α‐smooth‐muscle actin (α‐SMA)‐positive cells. In all primary tumors and liver metastases with desmoplastic growth pattern, TIMP‐1 mRNA was primarily found in α‐SMA‐positive myofibroblasts located at the invasive front. Some α‐SMA‐positive cells with TIMP‐1 mRNA were located adjacent to CD34‐positive endothelial cells, identifying them as pericytes. This indicates that TIMP‐1 in primary tumors and liver metastases with desmoplastic growth pattern has dual functions; being an MMP‐inhibitor at the cancer periphery and involved in tumor‐induced angiogenesis in the pericytes. In the liver metastases with pushing or replacement growth patterns, TIMP‐1 was primarily expressed by activated hepatic stellate cells at the metastasis/liver parenchyma interface. These cells were located adjacent to CD34‐positive endothelial cells, suggesting a function in tumor‐induced angiogenesis. We therefore conclude that TIMP‐1 expression is growth pattern dependent in colorectal cancer liver metastases. © 2015 The Authors. Molecular Carcinogenesis published by Wiley Periodicals, Inc.

• TIMP-1
• angiogenesis
• desmoplasia
• hepatic metastasis
• histology