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This file contains 10 supplemental figures. Supplemental Figure 1. JMJD3 is highly expressed in HCC tissues and cancer cell lines. Supplemental Figure 2. JMJD3 promotes proliferative capacity of HCC cells both in vivo. Supplemental Figure 3. JMJD3 promotes proliferative, EMT, migratory, and invasive capacities of HCC cells. Supplemental Figure. 4 JMJD3 promotes emergence of stem cell-like behavior in HCC cells. Supplemental Figure. 5 JMJD3 promotes emergence of stem cell-like behavior in HCC cells. Supplemental Figure. 6 JMJD3 regulates slug expression need its demethylase activity. Supplemental Figure. 7 JMJD3 regulates slug expression through H3K27 trimethylation Supplemental Figure. 8 JMJD3 regulates slug expression through H3K27 trimethylation Supplemental Figure. 9 Slug is a mediator for JMJD3-induced migration, invasion and in vivo metastasis capacity in HCC cells. Supplemental Figure. 10 Slug is a mediator for JMJD3-induced stemness capacity in HCC cells.

Supplemental Table 1. JMJD3 staining and clinicopathologic characteristics of 285 hepatocellular carcinoma patients. Supplemental Table 2. Primer sequences used for qRT-PCR and qChIP Supplemental Table 3. List of genes significantly differentially expressed after JMJD3 overexpression (fold change {greater than or equal to}5)

Supplemental Materials and Methods

The Jumonji domain–containing chromatin remodeling factor JMJD3 has important roles in development and cancer. Here, we report a pivotal role for JMJD3 in sustaining the phenotype of aggressive hepatocellular carcinomas. Expression levels of JMJD3 in clinical specimens of hepatocellular carcinoma correlated inversely with patient survival. In hepatocellular carcinoma cells, we found that enforcing its overexpression induced epithelial–mesenchymal transition (EMT), invasive migration, stem cell–like traits, and metastatic properties. Conversely, silencing JMJD3 in hepatocellular carcinoma cells overexpressing it inhibited these aggressive phenotypes. Mechanistically, JMJD3 modulated H3K27me3 in the SLUG gene promoter, a histone mark associated with active SLUG transcription. SLUG silencing blocked JMJD3-induced EMT, stemness, and metastasis. Furthermore, SLUG expression in hepatocellular carcinoma clinical specimens correlated positively with JMJD3 expression. Our results establish JMJD3 as a critical driver of hepatocellular carcinoma stem cell–like and metastatic behaviors, with implications for prognosis and treatment. Cancer Res; 76(22); 6520–32. ©2016 AACR.

This file contains 10 supplemental figures. Supplemental Figure 1. JMJD3 is highly expressed in HCC tissues and cancer cell lines. Supplemental Figure 2. JMJD3 promotes proliferative capacity of HCC cells both in vivo. Supplemental Figure 3. JMJD3 promotes proliferative, EMT, migratory, and invasive capacities of HCC cells. Supplemental Figure. 4 JMJD3 promotes emergence of stem cell-like behavior in HCC cells. Supplemental Figure. 5 JMJD3 promotes emergence of stem cell-like behavior in HCC cells. Supplemental Figure. 6 JMJD3 regulates slug expression need its demethylase activity. Supplemental Figure. 7 JMJD3 regulates slug expression through H3K27 trimethylation Supplemental Figure. 8 JMJD3 regulates slug expression through H3K27 trimethylation Supplemental Figure. 9 Slug is a mediator for JMJD3-induced migration, invasion and in vivo metastasis capacity in HCC cells. Supplemental Figure. 10 Slug is a mediator for JMJD3-induced stemness capacity in HCC cells.

The δ opioid receptor (DOR) is involved in the regulation of malignant transformation and tumor progression of hepatocellular carcinoma (HCC). However, regulation of the DOR in HCC remains poorly defined. We found that miR-874 was identified as a negative regulator of the DOR, which is a direct and functional target of miR-874 via its 3′ untranslated region (UTR). Moreover, miR-874 was downregulated in HCC and its expression was inversely correlated with DOR expression. Downregulation of miR-874 was also associated with larger tumor size, more vascular invasion, a poor TNM stage, poor tumor differentiation, and inferior patient outcomes. Functionally, overexpression of miR-874 in the HCC cell line SK-hep-1 inhibited cell growth, migration, in vitro invasion, and in vivo tumorigenicity. Furthermore, miR-874 overexpression suppressed the DOR, resulting in a downregulated epidermal growth factor receptor (EGFR) and extracellular signal-regulated kinase (ERK) phosphorylation. The EGFR activator—epidermal growth factor (EGF)—can rescue the proliferation and migration suppression induced by miR-874 overexpression, and the rescue effects of the EGF were blocked by an ERK inhibitor. Our study results suggest that miRNA-874 is a negative regulator of the DOR that can suppress tumor proliferation and metastasis in HCC by targeting the DOR/EGFR/ERK pathway, which may be a potential target for HCC treatment.

The Jumonji domain–containing chromatin remodeling factor JMJD3 has important roles in development and cancer. Here, we report a pivotal role for JMJD3 in sustaining the phenotype of aggressive hepatocellular carcinomas. Expression levels of JMJD3 in clinical specimens of hepatocellular carcinoma correlated inversely with patient survival. In hepatocellular carcinoma cells, we found that enforcing its overexpression induced epithelial–mesenchymal transition (EMT), invasive migration, stem cell–like traits, and metastatic properties. Conversely, silencing JMJD3 in hepatocellular carcinoma cells overexpressing it inhibited these aggressive phenotypes. Mechanistically, JMJD3 modulated H3K27me3 in the SLUG gene promoter, a histone mark associated with active SLUG transcription. SLUG silencing blocked JMJD3-induced EMT, stemness, and metastasis. Furthermore, SLUG expression in hepatocellular carcinoma clinical specimens correlated positively with JMJD3 expression. Our results establish JMJD3 as a critical driver of hepatocellular carcinoma stem cell–like and metastatic behaviors, with implications for prognosis and treatment. Cancer Res; 76(22); 6520–32. ©2016 AACR.

// Bo Tang 1,2 , Guangying Qi 2,3 , Fang Tang 1,2 , Shengguang Yuan 1,2 , Zhenran Wang 1,2 , Xingsi Liang 1,2 , Bo Li 1,2 , Shuiping Yu 1,2 , Jie Liu 1,2 , Qi Huang 1,2 , Yangchao Wei 1,2 , Run Zhai 1,2 , Biao Lei 1,2 , Hongping Yu 4 , Xingyuan Jiao 5 and Songqing He 1,2 1 Department of Hepatobiliary Surgery, Guilin Medical University, Affiliated Hospital, Guilin, Guangxi, People’s Republic of China 2 Laboratory of Liver Injury and Repair Molecular Medicine, Guilin Medical University, Guilin, Guangxi, People’s Republic of China 3 Department of Pathology and Physiopathology, Guilin Medical University, Guilin, Guangxi, People’s Republic of China 4 Department of Epidemiology and Statistics, School of Public Health, Guilin Medical College, Guilin, Guangxi, People’s Republic of China 5 Department of General Surgery, The First Affiliated Hospital, Sun Yat-Sen University, Guangzhou, People’s Republic of China Correspondence to: Songqing He, email: // Xingyuan Jiao, email: // Keywords : JARID1B, metastasis, epithelial-mesenchymal transition, invasion, PTEN Received : February 16, 2015 Accepted : March 10, 2015 Published : March 30, 2015 Abstract JARID1B is a member of the family of JmjC domain-containing proteins that removes methyl residues from methylated lysine 4 on histone H3 lysine 4 (H3K4). JARID1B has been proposed as an oncogene in many types of tumors; however, its role and underlying mechanisms in hepatocellular carcinoma (HCC) remain unknown. Here we show that JARID1B is elevated in HCC and its expression level is positively correlated with metastasis. In addition Kaplan-Meier survival analysis showed that high expression of JARID1B was associated with decreased overall survival of HCC patients. Overexpression of JARID1B in HCC cells increased proliferation, epithelial-mesenchymal transition, migration and invasion in vitro , and enhanced tumorigenic and metastatic capacities in vivo . In contrast, silencing JARID1B in aggressive and invasive HCC cells inhibited these processes. Mechanistically, we found JARID1B exerts its function through modulation of H3K4me3 at the PTEN gene promoter, which was associated with inactive PTEN transcription. PTEN overexpression blocked JARID1B-driven proliferation, EMT, and metastasis. Our results, for the first time, portray a pivotal role of JARID1B in stimulating metastatic behaviors of HCC cells. Targeting JARID1B may thus be a useful strategy to impede HCC cell invasion and metastasis.

Extraction of qualitative and quantitative information from large numbers of analytical signals is difficult with drifted baselines, particularly in multivariate analysis. Baseline drift obscures and "fuzzies" signals, and even deteriorates analytical results. In order to obtain accurate and clear results, some effective methods should be proposed and implemented to perform baseline correction before conducting further data analysis. However, most of the classic methods require user intervention or are prone to variability, especially with low signal-to-noise signals. In this study, a novel baseline correction algorithm based on quantile regression and iteratively reweighting strategy is proposed. This does not require user intervention and prior information, such as peak detection. The iteratively reweighting strategy iteratively changes weights of residuals between fitted baseline and original signals. After a series of tests and comparisons with several other popular methods, using various kinds of analytical signals, the proposed method is found to be fast, flexible, robust, and easy to use both in simulated and real datasets.

Run Zhai,1,2,* Fang Tang,3,* Jianhua Gong,1,2,* Jing Zhang,1,2 Biao Lei,1,2 Bo Li,2 Yangchao Wei,1,2 Xingsi Liang,1 Bo Tang,1,2 Songqing He1,2 1Laboratory of Liver Injury and Repair Molecular Medicine, Guilin Medical University, Guilin, Guangxi, People’s Republic of China; 2Department of Hepatobiliary Surgery, Affiliated Hospital, Guilin Medical University, Guilin, Guangxi, People’s Republic of China; 3Pathology Department, Affiliated Hospital, Guilin Medical University, Guilin, Guangxi, People’s Republic of China *These authors contributed equally to this work Abstract: Recent studies have shown that deubiquitination plays a key role in tumor progression, metastasis, resistance to chemotherapy drugs, and prognosis. In this study, we investigated the effects of the deubiquitinating enzyme USP22 on the expression of the drug-resistance genes BMI1 and EZH2 in hepatocellular carcinoma (HCC) and on prognosis. Downregulation of USP22 expression with interference ribonucleic acid in resistant HCC cell lines with high USP22 expression resulted in decreased BMI1 expression, but had no effect on EZH2 expression. USP22, BMI1, and EZH2 were highly expressed in HCC tissue, and the expression levels were positively correlated with tumor grade and clinical stage. Correlation analysis showed that USP22 expression was positively correlated with that of BMI1. Kaplan–Meier analysis showed that high levels of USP22 and BMI1 expression were associated with poor overall survival and relapse-free survival in all of the cases and in patients treated with transcatheter arterial chemoembolization. These results suggested that high levels of USP22 expression played an important role in drug resistance to chemotherapeutic drugs in HCC patients by upregulating the expression of BMI1; therefore, coexpression of USP22 and BMI1 may become a new predictor for HCC prognosis and may help guide clinical treatment. Keywords: hepatocellular carcinoma, USP22, BMI1, EZH2

Background Liver cancer is one of the main causes of cancer-related death in human. HOXA7 has been proved to be related with several cancers. Method The expression levels of HOXA7 were examined by Western blot, qRT-PCR or ICH. MTT was used to detect the proliferative rate of liver cancer cells. The invasive abilities were examined by matrigel and transwell assay. The metastatic abilities of liver cancer cells were revealed in BALB/c nude mice. Results In this report, we revealed that HOXA7 promoted metastasis of HCC patients. First, increased levels of HOXA7 were examined in liver cancer especially in metastatic liver cancer. Moreover, higher expression level of HOXA7 was associated with poorer prognosis of liver cancer patients. Overexpression of HOXA7 significantly enhanced proliferation, migration, invasion in vitro and tumor growth and metastasis in vivo meanwhile silencing HOXA7 significantly inhibited the aboves abilities of liver cancer cells. In this research, we identified that HOXA7 performed its oncogenic characteristics through activating Snail. Conclusion Collectively, we identify the critical role and possible mechanism of HOXA7 in metastasis of liver cancer which suggest that HOXA7 may be a potential therapeutic target of liver cancer patients. Electronic supplementary material The online version of this article (doi:10.1186/s12943-016-0540-4) contains supplementary material, which is available to authorized users.

Xingsi Liang,1,2,* Bin Zhang,3,* Quan Chen,4,* Jing Zhang,1,5 Biao Lei,1,5 Bo Li,5 Yangchao Wei,1,5 Run Zhai,1,5 Zhiqing Liang,2 Songqing He,1,5 Bo Tang1,5 1Laboratory of Liver Injury and Repair Molecular Medicine, Guilin Medical University, 2Department of Infectious Diseases, Guilin Medical University, Affiliated Hospital, Guilin, Guangxi, 3Department of Oncology, The First Affiliated Hospital of Dalian Medical University, Dalian, Liaoning, 4Department of Anesthesiology, The First Affiliated Hospital of Liaoning Medical University, Jinzhou, Liaoning, 5Department of Hepatobiliary Surgery, Guilin Medical University, Affiliated Hospital, Guilin, Guangxi, People’s Republic of China *These authors contributed equally to this work Abstract: Characterized by its acute onset, critical condition, poor prognosis, and high mortality rate, severe acute pancreatitis (SAP) can cause multiple organ failure at its early stage, particularly acute lung injury (ALI). The pathogenesis of ALI is diffuse alveolar damage, including an increase in pulmonary microvascular permeability, a decrease in compliance, and invasion of many inflammatory cells. Corticosteroids are the main treatment method for ALI; however, the associated high toxicity and side effects induce pain in patients. Recent studies show that the effective components in many traditional Chinese medicines can effectively inhibit inflammation with few side effects, which can decrease the complications caused by steroid consumption. Based on these observations, the main objective of the current study is to investigate the effect of alpinetin, which is a flavonoid extracted from Alpinia katsumadai Hayata, on treating lung injury induced by SAP and to explore the mechanism underlying the alpinetin-mediated decrease in the extent of ALI. In this study, we have shown through in vitro experiments that a therapeutic dose of alpinetin can promote human pulmonary microvascular endothelial cell proliferation. We have also shown via in vitro and in vivo experiments that alpinetin upregulates aquaporin-1 and, thereby, inhibits tumor necrosis factor-α expression as well as reduces the degree of lung injury. Overall, our study shows that alpinetin alleviates SAP-induced ALI. The likely molecular mechanism includes upregulated aquaporin expression, which inhibits tumor necrosis factor-α and, thus, alleviates SAP-induced ALI. Keywords: alpinetin, aquaporin-1, severe acute pancreatitis, acute lung injury, HPMVEC, tumor necrosis factor-&alpha

// Yi Zhang 1, 2, * , Changwei Lin 3, * , Guoqing Liao 1 , Sheng Liu 1 , Jie Ding 4 , Fang Tang 5 , Zhenran Wang 5 , Xingsi Liang 5 , Bo Li 5 , Yangchao Wei 5 , Qi Huang 5 , Xuan Li 5 , Bo Tang 5 1 Department of Gastrointestinal Surgery, Xiangya Hospital, Central South University, 410008, PR China 2 Department of Oncological Surgery, Affiliated Hospital of Xuzhou Medical College, 221000, PR China 3 Department of Gastrointestinal Surgery, Third Xiangya Hospital, Central South University, 410008, PR China 4 Department of Gastrointestinal Surgery, Guizhou Provincial People's Hospital, 550000, PR China 5 Department of Hepatobiliary Surgery, Affiliated Hospital of Guilin Medical University, 541000, PR China * These authors have contributed equally to this work Correspondence to: Guoqing Liao, e-mail: liaoguoqing@medmail.com.cn Bo Tang, e-mail: dytangbo@163.com Keywords: miR-506, EZH2, proliferation, metastasis, colon cancer Received: March 27, 2015 Accepted: September 21, 2015 Published: October 03, 2015 ABSTRACT Increasing evidence reveals that aberrant expression of microRNA contributes to the development and progression of colon cancer, but the roles of microRNA-506 (miR-506) in colon cancer remain elusive. Here, we demonstrated that miR-506 was down-regulated in colon cancer tissue and cells and that miR-506 expression was inversely correlated with EZH2 expression, tumor size, lymph node invasion, TNM stage and metastasis. A high level of miR-506 identified patients with a favorable prognosis. In vitro and in vivo experiments confirmed that miR-506 inhibits the proliferation and metastasis of colon cancer, and a luciferase reporter assay confirmed that EZH2 is a direct and functional target of miR-506 via the 3′UTR of EZH2. The restoration of EZH2 expression partially reversed the proliferation and invasion of miR-506-overexpressing colon cancer cells. Moreover, we confirmed that the miR-506-EZH2 axis inhibits proliferation and metastasis by activating/suppressing specific downstream tumor-associated genes and the Wnt/β-catenin signaling pathway. Taking together, our study sheds light on the role of miR-506 as a suppressor for tumor growth and metastasis and raises the intriguing possibility that miR-506 may serve as a new potential marker for monitoring and treating colon cancer.

Liver ischemia-reperfusion injury (IRI) is a major cause of morbidity and mortality after resection surgery, liver transplantation, and hemorrhagic and septic shock. Mir-155 is upregulated by a broad range of inflammatory mediators, and it has been demonstrated to be involved in both innate and adaptive immune responses. However, the role of mir-155 in liver IRI has never been investigated. In this study, mir-155 deficiency protected mice from liver IRI, as shown by lower serum alanine aminotransferase (ALT) levels and Suzuki scores. Mir-155 deficiency results in the development of M2 macrophages, which respond to IR-induced innate immune stimulation by producing a regulatory inflammatory response with higher level of IL-10, but lower levels of TNF-α, IL-6, and IL-12p40. Mir-155 deficiency suppresses IL-17 expression, which contributes to the liver IRI development. In our further in vitro study, the results show that the Th17 differentiation is inhibited by SOCS1 overexpression and the promoted M2 macrophage development induced by mir-155 deficiency is abolished by SOCS1 knockdown. In conclusion, mir-155 deficiency attenuates liver IRI through upregulation of SOCS1, and this was associated with promoted M2 macrophage and inhibited Th17 differentiation.