FCN2 inhibits epithelial–mesenchymal transition-induced metastasis of hepatocellular carcinoma via TGF-β/Smad signaling

doi:10.1016/j.canlet.2016.05.007

Cancer Letters

Volume 378, Issue 2, 10 August 2016, Pages 80-86

https://doi.org/10.1016/j.canlet.2016.05.007 Get rights and content

Highlights

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FCN2 can inhibit HCC invasion and metastasis.
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FCN2 exerts its functions through inhibiting EMT.
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FCN2 can block EMT induced by TGFβ/Smad pathway in HCC.

Abstract

Hepatocellular carcinoma (HCC) is currently still a major cause of cancer-related deaths. Identifying early metastatic biomarkers and therapeutic targets for HCC is of great importance. Emerging evidence suggest that epithelial–mesenchymal transitions (EMTs) play important roles in tumor metastasis and recurrence. Understanding molecular mechanisms that regulate the EMT process is crucial for improving HCC. In this study, we find Ficolin-2 (FCN2) plays an essential role in metastasis and EMT of HCC. FCN2 expression is downregulated in HCC cells and tissues. Low level of FCN2 in HCCs is correlated with aggressive metastatic features, and would be a prognostic factor for overall disease-free survival of HCC patients. Ectopic expression of FCN2 markedly inhibits HCC cells migration, invasion as well as EMT in vitro and in vivo. Moreover, TGF-β is found contribute to the function of FCN2 in suppressing metastasis and EMT of HCC. Collectively, our data suggest that FCN2 may have prognostic value in HCC metastasis. Additionally, the FCN2/ TGF-β/EMT axis identified in this study provides novel insight into the mechanisms of HCC metastasis, which may facilitate the development of new therapeutics against HCC.

Introduction

Hepatocellular carcinoma (HCC), the third leading cause of cancer-related death globally, is one of the most common malignancies [1]. In spite of the progress in understanding, diagnosis and treatment, HCC remains a malignant tumor with a very low 5-year survival rate, which is predominantly attributed to recurrence and metastasis [2]. But the molecular mechanisms underlying HCC metastasis remain far from fully understood. Thus, it is of importance to discover the mechanism of HCC metastasis.

The epithelial–mesenchymal transition (EMT) is a reversible procedure in which epithelial cells alter into mesenchymal cells and transform their adhesion and migratory capacity with decrease of the epithelial biomarker E-cadherin and increase of mesenchymal markers vimentin and N-cadherin. EMT is currently reported having a significant role in the migration and invasion of various malignancies [3]. D.J. McConkey et al. and O.O. Ogunwobi and Liu reported that EMT could promote the metastasis of a wide variety of tumors, for example, hepatocellular carcinoma, colon carcinoma, bladdercancer, malignant melanoma, etc [4], [5]. Meanwhile, it is widely accepted that the activation of canonical TGF β signaling pathway is a critical molecular event to induce EMT [6].

Gene chip analysis of ten pairs of clinical HCC specimens with or without distant metastasis resected from HCC patients distinctly revealed that reduced gene F (collagen/fibrinogen domain containing lectin) 2 (FCN2) expression could be observed in HCCs having distant metastasis, which means there is an inverse correlation between gene FCN2 and HCC metastasis. In addition, there is a positive correlation between gene FCN2 and prognosis of HCC patients which includes overall survival (OS). Gene FCN2 locates in Chromosome 9 (9q34), whose product (ficolin-2/L-ficolin) is a lectin-complement pathway activator in normal human plasma, services a critical role in innate immunity and binds to infectious diseases [7]. Zhao Y et al. have reported that the biological function of FCN2 is that inhibits the entry of hepatitis C virus (HCV) at an early stage of viral infection [8]. Hoang et al. found ficolin-2 levels and FCN2 haplotypes conduced to hepatitis B virus (HBV) infection outcome in Vietnamese patients [9]. Moreover, it is reported FCN2 play crucial roles in various types of bacterial diseases, including, mycobacterium tuberculosis, gram-negative bacteria and gram-positive bacteria [10], [11], [12]. Although FCN2 has been proven to have opsonic properties and is be able to initiate complement by the lectin pathway; whether it contributes to tumor growth, or has other functions of metastasis in HCC, are still unknown.

TGF β signaling has been demonstrated as an important driver of EMT via activation of Smad complexes that translocate into the nucleus to change gene expression, which significantly enhances HCC invasiveness and metastasis [13], [14]. In the present study, we report that reduced FCN2 expression correlated well with the metastasis of HCC by induction of TGF β signaling pathway to stimulate EMT process. These data forcefully suggested that FCN2 might be a critical antioncogene in hepatocellular carcinogenesis and metastasis.

Section snippets

Patients, tissue samples and gene chips analysis

Archival HCC tissues were obtained from patients at the First Affiliated Hospital of Harbin Medical University between 2008 and 2013. Informed consent was obtained from each patient prior to biopsy or surgery and the ethical approval for the use of human subjects was obtained from the Research Ethics Committee of the First Affiliated Hospital of Harbin Medical University. 10 pairs of clinical specimens with or without distant metastasis were under gene chips analysis from CapitalBio (Beijing,

Decreased FCN2 expression in Human HCCs predicts poor prognosis

In all, 10 pairs HCC tissues were selected from patients who underwent surgical resections in the First Affiliated Hospital of Harbin Medical University, Harbin, China from 2008 to 2013. By following up these patients, based on the recurrence, metastasis and survival, the HCC tissues were divided into two groups: distant metastasis being observed within 6 months (5/10) and not being detected within 5 years (5/10). Then gene chip was performed (Fig. 1A, Fig. S1A) to search the potential genes

Discussion

Active metastasis contributes to poor survival and high mortality of HCC, frequently accompanied by the process of EMT in which typical epithelial phenotype cells transform into spindle-shaped mesenchymal phenotype cells. In the process of EMT, epithelial cells lose their cell polarity and adhesion, and adopt mesenchymal characteristics by altering migratory capacity, which is closely related to tumor invasiveness, metastasis, and chemotherapy resistance [20]. It has been indicated by

Conflict of interest

The authors declare no conflict of interest.

Acknowledgments

This study was supported by the Graduate Student Research Innovation Foundation of Harbin Medical University (Grant No. YJSCX2014-34HYD), Changjiang Scholars and Innovative Research Team in University (Grant No. IRT1122), the National Natural Science Foundation of China (No. 81272705 and No. 81301807), the Heilongjiang Province Nature Science Foundation (LC2013C31), Innovative Research Team (in Science and Technology) in Higher Educational Institutions of Heilongjiang Province (2009td06),

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¹: These authors contributed equally to this work.

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Original ArticlesFCN2 inhibits epithelial–mesenchymal transition-induced metastasis of hepatocellular carcinoma via TGF-β/Smad signaling

Highlights

Abstract

Introduction

Section snippets

Patients, tissue samples and gene chips analysis

Decreased FCN2 expression in Human HCCs predicts poor prognosis

Discussion

Conflict of interest

Acknowledgments

Lancet (London, England)

Urol. Oncol

Crit. Rev. Oncol. Hematol

FEBS Lett

J. Hepatol

Cancer Lett

FEBS Lett

Cell

TRIM proteins and cancer

Nat. Rev. Cancer

An autocrine loop between TGF-beta1 and the transcription factor brachyury controls the transition of human carcinoma cells into a mesenchymal phenotype

Mol. Cancer Ther

Forkhead box Q1 promotes hepatocellular carcinoma metastasis by transactivating ZEB2 and VersicanV1 expression

Hepatology

Polymorphisms in the FCN2 gene determine serum variation and function of Ficolin-2

Hum. Mol. Genet

Ficolin-2 inhibits hepatitis C virus infection, whereas apolipoprotein E3 mediates viral immune escape

J. Immunol

Ficolin-2 levels and FCN2 haplotypes influence hepatitis B infection outcome in Vietnamese patients

PLoS ONE

Ficolin-2 defends against virulent Mycobacteria tuberculosis infection in vivo, and its insufficiency is associated with infection in humans

PLoS ONE

Original Articles
FCN2 inhibits epithelial–mesenchymal transition-induced metastasis of hepatocellular carcinoma via TGF-β/Smad signaling