• epithelial–mesenchymal transition
• intrahepatic cholangiocarcinoma
• septin 9
• vimentin
• ‘don't eat me’ signal

• Humans
• Cholangiocarcinoma/pathology
• Cholangiocarcinoma/metabolism
• Cholangiocarcinoma/immunology
• Cholangiocarcinoma/genetics
• Septins/metabolism
• Septins/genetics
• Bile Duct Neoplasms/pathology
• Bile Duct Neoplasms/metabolism
• Bile Duct Neoplasms/genetics
• Bile Duct Neoplasms/immunology
• Cell Line, Tumor
• Gene Expression Regulation, Neoplastic
• Epithelial-Mesenchymal Transition/genetics
• Vimentin/metabolism
• Vimentin/genetics
• Interferon-gamma/metabolism
• Signal Transduction

• For U1193 Institut National de la Santé et de la Recherche Médicale
• Ministère de l'Education Nationale
• Institut National de la Santé et de la Recherche Médicale
• Ministère de l'Education Nationale

• biliary epithelial cells
• cellular senescence
• cholangiocyte secretome
• cholangiopathies
• ductular reaction
• fibrosis
• hepatic carcinogenesis
• inflammation

• Cholangiocarcinoma
• Immunohistochemistry
• Microarray
• Prognosis

• Humans
• Prognosis
• Interleukin-6/metabolism
• Neoplasm Recurrence, Local/metabolism
• Cholangiocarcinoma/surgery
• Bile Duct Neoplasms/surgery
• Cadherins/analysis
• Cadherins/metabolism
• ErbB Receptors/analysis
• ErbB Receptors/metabolism
• Bile Ducts, Intrahepatic
• Immunoassay

• epithelial mesenchymal transformation
• laryngeal neoplasms
• polyploid giant cancer cells
• prognosis

• National Natural Science Foundation of China

• GBM
• YY1
• circular RNA circ_0001588
• miR-211-5p
• progression

• EMT
• Wnt signalling
• biomarker
• cancer stem cell-like property
• therapeutic target

Background: Glioma is the most common malignant tumor of the central nervous system, and often displays invasive growth. Recently, circular RNA (circRNA), which is a novel non-coding type of RNA, has been shown to play a vital role in glioma tumorigenesis. However, the functions and mechanism of lipocalin-2 (Lcn2)-derived circular RNA (hsa_circ_0088732) in glioma progression remain unclear.

Methods: We evaluated hsa_circ_0088732 expression by fluorescence in situ hybridization (FISH), Sanger sequencing, and PCR assays. Cell apoptosis was evaluated by flow cytometry and Hoechst 33258 staining. Transwell migration and invasion assays were performed to measure cell metastasis and viability. In addition, the target miRNA of hsa_circ_0088732 and the target gene of miR-661 were predicted by a bioinformatics analysis, and the interactions were verified by dual-luciferase reporter assays. RAB3D expression was analyzed by an immunochemistry assay, and E-cadherin, N-cadherin, and vimentin protein expression were examined by western blot assays. A mouse xenograft model was developed and used to analyze the effects of hsa_circ_0088732 on glioma growth in vivo.

Results: We verified that hsa_circ_0088732 is circular and highly expressed in glioma tissues. Knockdown of hsa_circ_0088732 induced glioma cell apoptosis and inhibited glioma cell migration, invasion, and epithelial-mesenchymal transition (EMT). We found that hsa_circ_0088732 negatively regulated miR-661 by targeting miR-661, and RAB3D was a target gene of miR-661. In addition, inhibition of miR-661 promoted glioma cell metastasis and suppressed cell apoptosis. Knockdown of RAB3D induced cell apoptosis and suppressed cell metastasis. Moreover, hsa_circ_0088732 accelerated glioma progression through its effects on the miR-661/RAB3D axis. Finally, results from a mouse xenograft model confirmed that knockdown of hsa_circ_0088732 induced miR-661 expression, resulting in suppression of RAB3D expression and inhibition of tumor growth in vivo.

Conclusion: We demonstrated that hsa_circ_0088732 facilitated glioma progression by sponging miR-661 to increase RAB3D expression. This study provides a theoretical basis for understanding the development and occurrence of glioma, as well as for the development of targeted drugs.

• EMT
• RAB3D
• glioma
• hsa_circ_0088732
• miR-661

• EMT
• ICC
• immunohistochemistry
• prognosis
• survival analysis

In many types of solid tumours, the aberrant expression of the cell adhesion molecule N-cadherin is a hallmark of epithelial-to-mesenchymal transition, resulting in the acquisition of an aggressive tumour phenotype. This transition endows tumour cells with the capacity to escape from the confines of the primary tumour and metastasise to secondary sites. In this review, we will discuss how N-cadherin actively promotes the metastatic behaviour of tumour cells, including its involvement in critical signalling pathways which mediate these events. In addition, we will explore the emerging role of N-cadherin in haematological malignancies, including bone marrow homing and microenvironmental protection to anti-cancer agents. Finally, we will discuss the evidence that N-cadherin may be a viable therapeutic target to inhibit cancer metastasis and increase tumour cell sensitivity to existing anti-cancer therapies.

• Cancer
• Haematological malignancies
• Metastasis
• N-cadherin
• Therapeutic target

The functional roles and clinical significances of miR-590-3p in ICC remain unclear. In the current study, we investigated the expression of miR-590-3p in tissues and sera of ICC by real-time quantitative polymerase chain reaction. We found miR-590-3p was significantly down-regulated in the sera and tissues of ICC patients, especially in those patients with lymph node metastasis or distant metastasis. AUC curves and Cox proportional hazards mode revealed serum miR-590-3p could be novel diagnostic and prognostic biomarker for ICC patients. MiR-590-3p dramatically suppressed epithelial-mesenchymal transition, cell migration, and invasion of ICC cells. SIP1 was identified as direct and functional target of miR-590-3p in ICC cells by luciferase assays. Finally, we found SIP1 expression was inversely correlated with miR-590-3p and closely related to diminished survival in ICC patients. These findings reveal functional and mechanistic roles of miR-590-3p and EMT activator SIP1 in the pathogenesis of ICC.

• EMT
• SIP1
• intrahepatic cholangiocarcinoma
• metastasis
• miR-590-3p

Signal transducer and activator of transcription 3 (STAT3) promotes tumor progression in many types of cancer. In this study, we analyzed the prognostic value of this marker in human intrahepatic cholangiocarcinoma (ICC). Using real-time PCR, western blot and immunohistochemistry assays, we found that STAT3 is overexpressed in ICC patients. STAT3 expression correlated with several clinicopathological features, including tumor size, pathological satellite, vascular invasion, undifferentiated-type histology, lymph node metastasis and TNM stage in two independent cohorts of ICC patients. Patients with high STAT3 levels had a poor prognosis in terms of overall survival (OS) and disease-free survival (DFS). Multivariate survival analysis indicated that STAT3 is an independent prognostic factor for OS and DFS. Furthermore, we observed that STAT3 overexpression promotes the invasion, metastasis and proliferation of ICC cells in vitro and in vivo, and also promotes STAT3 phosphorylation. These findings suggest that STAT3 expression correlated negatively with surgical outcome and inhibition of STAT3 expression may constitute a novel target for the treatment of ICC patients.

• STAT3
• intrahepatic cholangiocarcinoma
• metastasis
• prognosis
• proliferation

• Cholangiocarcinoma
• EMT
• KITENIN

Periostin, a secreted matricellular protein, has been reported to induce epithelial‐mesenchymal transition (EMT), which increases motility and invasiveness in various epithelial cancer cells. Periostin is also overexpressed in intrahepatic cholangiocarcinoma (ICC) and suggested to be a biomarker for tumor progression and poor prognosis; however, its functional role in ICC is not fully understood. Here, we investigated whether periostin influences malignant potential through the induction of EMT in ICC. Analyses of surgical resected ICC specimens revealed that the gene expression of periostin was significantly higher in ICC tumors than in adjacent nontumor liver tissues and was closely correlated with the expression of mesenchymal markers, including N‐cadherin, vimentin, and fibronectin. However, the expression level of periostin varied in each case. Consistently, the expression of periostin in HuH28 (an undifferentiated ICC cell) was markedly higher than in HuCCT‐1 (a moderately differentiated ICC cell). In addition, high‐level secretion of periostin into culture media was observed in HuH28 but not in HuCCT‐1. To identify the biological significance of periostin in EMT, gene silencing of periostin by small interfering RNA was performed in HuH28 cells. Periostin knockdown in HuH28 cells significantly down‐regulated mesenchymal markers and up‐regulated epithelial markers, suggesting the reversal of EMT, namely mesenchymal‐epithelial transition. Along with these changes, cell proliferation was significantly suppressed by 52%. In addition, cell migration and invasion were significantly suppressed by 62% and 61%, respectively, with reduced gene expression of matrix metalloproteinase 2. Interestingly, chemosensitivity to gemcitabine was also significantly improved by periostin depletion. Conclusion: Periostin plays an important role in the regulation of malignant potential through EMT and is suggested to be a novel target for the treatment of ICC. (Hepatology Communications 2017;1:1099–1109)

Transforming growth factor-β (TGF-β) plays a paradoxical role in cancer: it suppresses proliferation at early stages but promotes metastasis at late stages. This cytokine is upregulated in cholangiocarcinoma and is implicated in cholangiocarcinoma invasion and metastasis. Here we investigated the roles of non-Smad pathway (ERK1/2) and Smad in TGF-β tumor promoting and suppressing activities in intrahepatic cholangiocarcinoma (ICC) cells.

TGF-β1 effects on proliferation, invasion and migration of ICC cells, KKU-M213 and/or HuCCA-1, were investigated using MTT, colony formation, in vitro Transwell and wound healing assays. Levels of mRNAs and proteins/phospho-proteins were measured by quantitative (q)RT-PCR and Western blotting respectively. E-cadherin localization was examined by immunofluorescence and secreted MMP-9 activity was assayed by gelatin zymography. The role of ERK1/2 signaling was evaluated by treating cells with TGF-β1 in combination with MEK1/2 inhibitor U0126, and that of Smad2/3 and Slug using siSmad2/3- and siSlug-transfected cells.

h-TGF-β1 enhanced KKU-M213 cell invasion and migration and induced epithelial-mesenchymal transition as shown by an increase in vimentin, Slug and secreted MMP-9 levels and by a change in E-cadherin localization from membrane to cytosol, while retaining the cytokine’s ability to attenuate cell proliferation. h-TGF-β1 stimulated Smad2/3 and ERK1/2 phosphorylation, and the MEK1/2 inhibitor U0126 attenuated TGF-β1-induced KKU-M213 cell invasion and MMP-9 production but moderately enhanced the cytokine growth inhibitory activity. The latter effect was more noticeable in HuCCA-1 cells, which resisted TGF-β-anti-proliferative activity. Smad2/3 knock-down suppressed TGF-β1 ability to induce ERK1/2 phosphorylation, Slug expression and cell invasion, whereas Slug knock-down suppressed cell invasion and vimentin expression but marginally affected ERK1/2 activation and MMP-9 secretion. These results indicate that TGF-β1 activated ERK1/2 through Smad2/3 but not Slug pathway, and that ERK1/2 enhanced TGF-β1 tumor promoting but repressed its tumor suppressing functions.

Inhibiting ERK1/2 activation attenuates TGF-β1 tumor promoting effect (invasion) but retains its tumor suppressing role, thereby highlighting the importance of ERK1/2 in resolving the TGF-β paradox switch.

The online version of this article (doi:10.1186/s12935-017-0454-2) contains supplementary material, which is available to authorized users.

• Cholangiocarcinoma
• ERK1/2
• Invasion
• Slug
• Smad2/3
• TGF-β1

• Chemoresistance
• Cholangiocarcinoma
• Epithelial-mesenchymal transition
• Invasiveness
• Tumor microenvironment

• Bmi-1
• EMT
• PTEN
• Sal-like protein 4
• intrahepatic cholangiocarcinoma
• metastasis

• CHOLANGIOCARCINOMA
• IMMUNOHISTOCHEMISTRY
• LIVER CANCER

• Aged
• Bile Duct Neoplasms/metabolism
• Bile Duct Neoplasms/mortality
• Bile Duct Neoplasms/pathology
• Bile Ducts, Intrahepatic/metabolism
• Bile Ducts, Intrahepatic/pathology
• Cadherins/metabolism
• Cholangiocarcinoma/metabolism
• Cholangiocarcinoma/mortality
• Cholangiocarcinoma/pathology
• Female
• Humans
• Male
• Middle Aged
• Prognosis
• Survival Rate
• Vimentin/metabolism
• Zinc Finger E-box-Binding Homeobox 1/metabolism

• Epithelial-mesenchymal transition
• Intrahepatic cholangiocarcinoma
• Invasion
• Migration
• WAVE3

• Cell Line, Tumor
• Cell Movement/physiology
• Cell Proliferation/physiology
• Cholangiocarcinoma/metabolism
• Epithelial-Mesenchymal Transition/physiology
• Gene Expression Regulation, Neoplastic/physiology
• Humans
• Neoplasm Invasiveness/pathology
• Wiskott-Aldrich Syndrome Protein Family/genetics
• Wiskott-Aldrich Syndrome Protein Family/metabolism

Cholangiocarcinomas are neoplasms that involve the epithelial cells of the bile duct, also known as cholangiocytes. This disease is difficult to diagnose early, as most symptoms present late in the disease. In addition, the specific anatomic position can cause periductal extension and result in a very low radical excision rate and a very poor prognosis. Improved understanding of the features underlying the onset of cholangiocarcinoma and its carcinogenic mechanism may lead to early diagnosis and better prognosis. With the development of molecular biology, much has been learned about oncogenes, tumor-suppressor genes, DNA methylation, microRNAs, and the molecular mechanisms of tumor invasion and metastasis. Based on our research and others, this review article will discuss the current status and prospects of early diagnosis of cholangiocarcinoma.

• polyploid giant cancer cells
• breast cancer
• epithelial-mesenchymal transition
• metastasis

• Adult
• Breast Neoplasms/chemically induced
• Breast Neoplasms/genetics
• Breast Neoplasms/pathology
• Cadherins/biosynthesis
• Cadherins/genetics
• Cell Line, Tumor
• Cobalt/toxicity
• Epithelial-Mesenchymal Transition/drug effects
• Female
• Gene Expression Regulation, Neoplastic/drug effects
• Giant Cells/drug effects
• Giant Cells/pathology
• Humans
• Lymphatic Metastasis
• MCF-7 Cells
• Middle Aged
• Neoplasm Invasiveness/genetics
• Neoplasm Invasiveness/pathology
• Polyploidy
• Vimentin/biosynthesis
• Vimentin/genetics

In addition to its well-established role in embryo development, epithelial-to-mesenchymal transition (EMT) has been proposed as a general mechanism favoring tumor metastatization in several epithelial malignancies. Herein, we review the topic of EMT in cholangiocarcinoma (CCA), a primary liver cancer arising from the epithelial cells lining the bile ducts (cholangiocytes) and characterized by an abundant stromal reaction. CCA carries a dismal prognosis, owing to a pronounced invasiveness and scarce therapeutic opportunities. In CCA, several reports indicate that cancer cells acquire a number of EMT biomarkers and functions. These phenotypic changes are likely induced by both autocrine and paracrine signals released in the tumor microenvironment (cytokines, growth factors, morphogens) and intracellular stimuli (microRNAs, oncogenes, tumor suppressor genes) variably associated with specific disease mechanisms, including chronic inflammation and hypoxia. Nevertheless, evidence supporting a complete EMT of neoplastic cholangiocytes into stromal cells is lacking, and the gain of EMT-like changes by CCA cells rather reflects a shift towards an enhanced pro-invasive phenotype, likely induced by the tumor stroma. This concept may help to identify new biomarkers of early metastatic behavior along with potential therapeutic targets.

• cancer-associated fibroblast
• cholangiocarcinoma
• cholangiocyte
• epithelial-to-mesenchymal transition
• invasiveness
• metastatization
• tumor reactive stroma

• R01 DK079005 NIDDK NIH HHS

The set definition of distal cholangiocarcinomas and adenocarcinomas of the pancreatic head is challenged by their close anatomical relation, similar growth pattern, and corresponding therapeutic outcome. They show a mutual development during embryologic organ formation and share phenotypic characteristics. This review will highlight the similarities with regard to the common origin of their primary organs, histopathological similarities, and modern clinical management. Thus, we propose to subsume those entities under a common superfamily.

• Bile duct neoplasm
• cholangiocarcinoma
• pancreatic neoplasm

• E-cadherin
• Oral squamous cell carcinoma
• epithelial-mesenchymal transition
• immunohistochemistry
• vimentin

• Adult
• Aged
• Aged, 80 and over
• Biomarkers, Tumor/analysis
• Cadherins/biosynthesis
• Carcinoma, Squamous Cell/pathology
• Epithelial-Mesenchymal Transition/physiology
• Female
• Humans
• Immunohistochemistry
• Lymphatic Metastasis/pathology
• Male
• Middle Aged
• Mouth Neoplasms/pathology
• Vimentin/biosynthesis

• Bmi1
• Twist
• cancers: biology
• diagnosis and therapy
• epithelial-mesenchymal transition
• pancreatic neoplasms: biology and therapy

• biliary tree
• cholangiocarcinoma
• dyskerin
• histopathology
• immunohistochemistry

• Biliary cancers
• Biomarkers
• Gene fusions
• Gene mutations
• Liver cancers

Intrahepatic cholangiocarcinoma is macroscopically classified into three subtypes, mass-forming-type, periductal infiltrating-type, and intraductal growth-type. Each subtype should be preoperatively differentiated to perform the valid surgical resection. Recent researches have revealed the clinical, radiologic, pathobiological characteristics of each subtype. We reviewed recently published studies covering various aspects of intrahepatic cholangiocarcinoma (ICC), focusing especially on the macroscopic subtypes and stem cell features to better understand the pathophysiology of ICC and to establish the valid therapeutic strategy.

• Intrahepatic cholangiocarcinoma
• Combined hepatocellular-cholangiocarcinoma
• Hepatic progenitor cells
• Macroscopic subtype

• Cholangiocarcinoma
• E-cadherin
• EGF
• EGFR
• Invasion
• Migration

• Bile Duct Neoplasms/diagnosis
• Bile Duct Neoplasms/metabolism
• Bile Duct Neoplasms/pathology
• Bile Ducts, Intrahepatic/metabolism
• Bile Ducts, Intrahepatic/pathology
• Cadherins/metabolism
• Cholangiocarcinoma/diagnosis
• Cholangiocarcinoma/metabolism
• Cholangiocarcinoma/pathology
• Epithelial-Mesenchymal Transition
• Gene Expression Regulation, Neoplastic
• Humans
• Prognosis
• Snail Family Transcription Factors
• Transcription Factors/metabolism
• Vimentin/metabolism
• beta Catenin/metabolism

• Bladder cancer cells
• EMT
• ILK
• Metastasis
• Ribonuclease inhibitor

Acquired resistance towards sorafenib treatment was found in HCC patients, which results in poor prognosis. To investigate the enhanced metastatic potential of sorafenib resistance cells, sorafenib-resistant (SorR) cell lines were established by long-term exposure of the HCC cells to the maximum tolerated dose of sorafenib. Cell proliferation assay and qPCR of ABC transporter genes (ABCC1-3) were first performed to confirm the resistance of cells. Migration and invasion assays, and immunoblotting analysis on the expression of epithelial to mesenchymal transition (EMT) regulatory proteins were performed to study the metastatic potential of SorR cells. The expression of CD44 and CD133 were studied by flow cytometry and the gene expressions of pluripotency factors were studied by qPCR to demonstrate the enrichment of cancer stem cells (CSCs) in SorR cells. Control (CTL) and SorR cells were also injected orthotopically to the livers of NOD-SCID mice to investigate the development of lung metastasis. Increased expressions of ABCC1-3 were found in SorR cells. Enhanced migratory and invasive abilities of SorR cells were observed. The changes in expression of EMT regulatory proteins demonstrated an activation of the EMT process in SorR cells. Enriched proportion of CD44⁺ and CD44⁺CD133⁺ cells were also observed in SorR cells. All (8/8) mice injected with SorR cells demonstrated lung metastasis whereas only 1/8 mouse injected with CTL cells showed lung metastasis. HCC cells with sorafenib resistance demonstrated a higher metastatic potential, which may be due to the activated EMT process. Enriched CSCs were also demonstrated in the sorafenib resistant cells. This study suggests that advanced HCC patients with acquired sorafenib resistance may have enhanced tumor growth or distant metastasis, which raises the concern of long-term sorafenib treatment in advanced HCC patients who have developed resistance of sorafenib.

• ATP-Binding Cassette Transporters/metabolism
• Animals
• Antineoplastic Agents/pharmacology
• Carcinoma, Hepatocellular/drug therapy
• Carcinoma, Hepatocellular/metabolism
• Carcinoma, Hepatocellular/pathology
• Cell Line, Tumor
• Drug Resistance, Neoplasm
• Heterografts
• Humans
• Liver Neoplasms/drug therapy
• Liver Neoplasms/metabolism
• Liver Neoplasms/pathology
• Mice
• Neoplasm Metastasis
• Neoplasm Proteins/metabolism
• Neoplasm Transplantation
• Niacinamide/analogs & derivatives
• Niacinamide/pharmacology
• Phenylurea Compounds/pharmacology
• Sorafenib

• Animals
• Cholangiocarcinoma/genetics
• Cholangiocarcinoma/pathology
• Cholangiocarcinoma/physiopathology
• Cholangiocarcinoma/therapy
• Disease Models, Animal
• Humans
• Risk Factors