The interaction between T cell immunoglobulin-and mucin-domain-containing molecule (Tim-3) expressed on T helper 1 (Th1) cells, and its ligand, galectin-9, negatively regulates Th1-mediated immune responses. However, it is poorly understood if and how the Tim-3/ galectin-9 signaling pathway is involved in immune escape in patients with hepatocellular carcinoma (HCC). Here we studied the expression, function, and regulation of the Tim-3/ galectin-9 pathway in patients with hepatitis B virus (HBV)-associated HCC. We detected different levels of galectin-9 expression on antigen-presenting cell (APC) subsets including Kupffer cells (KCs), myeloid dendritic cells (DCs), and plasmacytoid DCs in HCC. The highest galectin-9 expression was on KCs in HCC islets, not in the adjacent tissues. Furthermore, Tim-3 expression was increased on CD4 1 and CD8 1 T cells in HCC as compared to the adjacent tissues, and Tim-3 1 T cells were replicative senescent and expressed surface and genetic markers for senescence. Interestingly, tumor-infiltrating T-cell-derived interferon (IFN)-c stimulated the expression of galectin-9 on APCs in the HCC microenvironment. Immunofluorescence staining revealed a colocalization of Tim-3 1 T cells and galectin-91 KCs in HCC. Functional studies demonstrated that blockade of the Tim-3/ galectin-9 signaling pathway importantly increased the functionality of tumor-infiltrating Tim-3 1 T cells as shown by increased T-cell proliferation and effector cytokine production. Finally, we show that the numbers of Tim-3 1 tumor-infiltrating cells were negatively associated with patient survival. Conclusion: Our work demonstrates that the Tim-3/galectin-9 signaling pathway mediates T-cell senescence in HBV-associated HCC. The data suggest that this pathway could be an immunotherapeutic target in patients with HBV-associated HCC. (HEPATOLOGY 2012;56:1342-1351 H epatocellular carcinoma (HCC) is one of the most common cancers. More than 80% of patients are not candidates for curative treatments with the final diagnosis, and are linked to chronic infection with the hepatitis B (HBV) or hepatitis C (HCV) viruses based on different regions.

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Expression of B7 costimulatory molecules represents an important compartment of immune response of epithelial cells following microbial infection. We reported here that the protozoan parasite Cryptosporidium parvum induced B7-H1 expression in cultured human cholangiocytes. Induced expression of B7-H1 was identified in cells after exposure to infective C. parvum parasite or parasite lysate. Interestingly, microRNA-513 (miR-513) level was reduced in cells after exposure to C. parvum, resulting in a relief of 3′-untranslated region-mediated translational suppression of B7-H1. Overexpression of miR-513 through transfection of miR-513 precursor inhibited C. parvum-induced B7-H1 protein expression. Moreover, enhanced apoptotic cell death was identified in activated human T cells following co-culture with C. parvum-infected cholangiocytes. The apoptosis of activated T cells was partially blocked by a neutralizing antibody to B7-H1 or transfection of cholangiocytes with miR-513 precursor. These data suggest a role of miR-513 in regulating B7-H1 expression by cholangiocytes in response to C. parvum infection.

Ion channels have been suggested to be important in the development and progression of tumors, however, chloride channels have rarely been analyzed in tumorigenesis. More recently, transmembrane protein with unknown function 16A (TMEM16A), hypothesized to be a candidate calcium‑activated Cl‑ channel, has been found to be overexpressed in a number of tumor types. Although several studies have implicated the overexpression of TMEM16A in certain tumor types, the exact role of TMEM16A in gliomas and the underlying mechanisms in tumorigenesis, remain poorly understood. In the present study, the role of TMEM16A in gliomas and the potential underlying mechanisms were analyzed. TMEM16A was highly abundant in various grades of gliomas and cultured glioma cells. Knockdown of TMEM16A suppressed cell proliferation, migration and invasion. Furthermore, nuclear factor‑κB (NF‑κB) was activated by overexpression of TMEM16A. In addition, TMEM16A regulated the expression of NF‑κB‑mediated genes, including cyclin D1, cyclin E and c‑myc, involved in cell proliferation, and matrix metalloproteinases (MMPs)‑2 and MMP‑9, which are associated with the migration and invasion of glioma cells. Collectively, results of the present study provide evidence for the involvement of TMEM16A in gliomas and the potential mechanism through which TMEM16A promotes glioma formation.

AAV producer cell lines can be readily scaled to meet the needs of clinical trials. One 500 L bioreactor of these producer cells can produce the equivalent of 2500 high capacity roller bottles or 25 000 T-175 tissue culture flasks.

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Jumonji domain-containing protein 6 (JMJD6), a histone arginine demethylase, plays a multifaceted and significant role in embryonic development and cancer progression. However, the function of JMJD6 and its precise mechanism in regulating hepatocellular carcinoma (HCC) remain unknown. Here, we show that aberrant JMJD6 overexpression is associated with poor prognosis and aggressive characteristics of HCC. In hepatoma cell lines, we demonstrated that knockdown of JMJD6 inhibited hepatoma cell migration and proliferation. JMJD6 overexpression displays the opposite effects. Interestingly, JMJD6 regulates hepatoma cell cycle and apoptosis progression. Moreover, there was a positive correlation between cell cycle regulatory protein CDK4 and JMJD6 level. Mechanism analysis suggested JMJD6 promotes CDK4 expression by directly targeting to its promoter, and interacts with PCAF to regulate the histone modifications on the promoter of CDK4. Furthermore, we found that inhibiting CDK4 abolished the ability of JMJD6 in enhancing cell proliferation. Taken together, for the first, we demonstrated that JMJD6 is critically involved in HCC carcinogenesis, and indicated that JMJD6 may be a novel potential biomarker for HCC.

MicroRNAs have been reported to be closely related to the development of human lung cancers. However, the functions of microRNAs in non-small cell lung cancer (NSCLC) remain largely undefined. Here, we investigated the role of microRNA-193b (miR-193b) in NSCLC. Our data showed that miR-193b was markedly down-regulated in NSCLC cancer tissues compared with adjacent normal tissues. The NSCLC cell line (A549) transfected with the miR-193b exhibited significantly decreased proliferation, migration, and invasion capacities when compared with the control cells. In contrast, inhibition of miR-193b increased the proliferation, migration, and invasion of A549 cells. Moreover, miR-193b repressed the expressions of cyclin D1 and urokinase-type plasminogen activator in A549 cells. These data suggest that miR-193b is a tumor suppressor in NSCLC.