Xiaoming Lu

Tim‐3/galectin‐9 signaling pathway mediates T‐cell dysfunction and predicts poor prognosis in patients with hepatitis B virus‐associated hepatocellular carcinoma

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+ and CD8+ T cells in HCC as compared to the adjacent tissues, and Tim‐3+ T cells were replicative senescent and expressed surface and genetic markers for senescence. Interestingly, tumor‐infiltrating T‐cell‐derived interferon (IFN)‐γ stimulated the expression of galectin‐9 on APCs in the HCC microenvironment. Immunofluorescence staining revealed a colocalization of Tim‐3+ T cells and galectin‐9+ 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+ T cells as shown by increased T‐cell proliferation and effector cytokine production. Finally, we show that the numbers of Tim‐3+ 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)

Expression of SATB1 and heparanase in gastric cancer and its relationship to clinicopathologic features

Cheng C, Lu X, Wang G, Zheng L, Shu X, Zhu S, Liu K, Wu K, Tong Q. Expression of SATB1 and heparanase in gastric cancer and its relationship to clinicopathologic features. APMIS 2010; 118: 855–63.

SATB1 is an independent prognostic marker for gastric cancer in a Chinese population

Special AT-rich sequence binding protein 1 (SATB1), a new type of gene regulator, has been reported to express in various human cancers and may associate with the malignant potential. However, there are few data available on SATB1 expression and its relationship to tumor progression in gastric cancer. The current study was designed to examine the SATB1 expression in gastric cancer and to correlate it with clinical outcome. SATB1 expression was studied by qRT-PCR, western blotting and immunohistochemistry, respectively. The correlations between SATB1 expression and clinicopathological factors were statistically analyzed. SATB1 mRNA expression was significantly up-regulated in gastric cancer specimens as compared with that in normal tissues (P<0.001). Overexpression of SATB1 protein was observed in gastric cancer cell lines by western blotting. Fifty-three (44.9%) cases showed positive staining for the SATB1 proteins by immunohistochemistry. The expression of SATB1 mRNA agreed well with the western blotting and immunohistochemical findings (P<0.001). SATB1 expression was positively correlated with age, depth of invasion, lymph node metastasis, distant metastasis and TNM stage. Moreover, Kaplan-Meier analysis revealed that SATB1 overexpression was associated with a significantly worse survival (P<0.001). Further multivariable analysis indicated that SATB1 expression was an independent prognostic indicator for gastric cancer (P=0.023). In summary, overexpression of SATB1 correlated with metastatic potential of human gastric cancer and would be a novel independent prognostic marker for predicting the outcome of gastric cancer.

Co-delivery of doxorubicin and SATB1 shRNA by thermosensitive magnetic cationic liposomes for gastric cancer therapy.

In previous a study, we had developed a novel thermosensitive magnetic delivery system based on liposomes. This study aimed to evaluate the efficiency of this system for the co-delivery of both drugs and genes to the same cell and its anti-tumor effects on gastric cancer. Doxorubicin (DOX) and SATB1 shRNA vector were loaded into the co-delivery system, and in vitro DOX thermosensitive release activity, targeted gene silencing efficiency, targeted cellular uptake, in vitro cytotoxicity, as well as in vivo anti-tumor activity were determined. The results showed that this co-delivery system had desirable targeted delivery efficacy, DOX thermosensitive release and SATB1 gene silencing. Moreover, the co-delivery of DOX and SATB1 shRNA exhibited enhanced activity to inhibit gastric cancer cell growth in vitro and in vivo, compared to single delivery. In conclusion, the novel thermosensitive magnetic drug and gene co-delivery system has promising application in combined chemotherapy and gene therapy for gastric cancer.

Conversion of intratumoral regulatory T cells by human gastric cancer cells is dependent on transforming growth factor‐β1

Regulatory T cells (Treg) inhibits immune responses mediated by T cells. This study aimed to investigate whether Treg are accumulated in human gastric cancer tissue and the mechanism of Treg induction by gastric cancer cells.

Increased expressions of SATB1 and S100A4 are associated with poor prognosis in human colorectal carcinoma.

This study was designed to explore the correlation between expressions of SATB1 and S100A4 and their relationships to the clinicopathologic parameters of colorectal carcinoma (CRC). Expressions of SATB1 and S100A4 were evaluated by immunohistochemistry in a cohort of 131 primary CRC patients undergone surgical resection from 2005 to 2007. SATB1 and S100A4 were positively expressed in 48.9% and 54.2% of CRC cases, respectively. SATB1 and S100A4 expressions in tumor tissues were significantly higher than those in the corresponding normal tissues. A positive correlation was observed between SATB1 and S100A4. Moreover, the levels of SATB1 and S100A4 were both significantly associated with invasion, lymph node status, and TNM stage of CRC, whereas S100A4 expression was also correlated with distant metastasis. Multivariate analysis revealed that SATB1 expression was an independent prognostic indicator for poor survival of CRC. Further survival analysis indicated that co‐expression of SATB1 and S100A4 suggested a worse 5‐year overall survival rate in CRC patients. Thus, combined analysis of SATB1 and S100A4 expressions may be valuable in determining the development and progression of CRC. Co‐expression of SATB1 and S100A4 is an unfavorable prognostic indicator and may be useful in the follow‐up of patients with CRC.

Construction and Characterization of a Novel Fusion Protein MG7-scFv/SEB against Gastric Cancer

Antibody-targeted superantigen has been developed into a new strategy to treat many malignant tumors. In this study, for specific targeting to gastric cancer cell, superantigen SEB (Staphylococcal Enterotoxin B) was genetically fused to the single-chain variable fragment of gastric carcinoma-associated antibody MG7(MG7-scFv) that recognizes the MG7 antigen frequently expressed in gastric cancer cell. The recombinant MG7-scFv/SEB fusion proteins are expressed in E. coli as inclusion bodies, and the purified MG7-scFv/SEB retains high binding affinity with gastric cancer cell SGC-7901 (positive MG7 antigen expression). When incubated with effector cell—peripheral blood mononuclear cells (PBMCs), MG7-scFv/SEB could effectively inhibit the proliferation and induce apoptosis of SGC-7901. After being treated with MG7-scFv/SEB, PBMCs remarkably increased the production of Th1 cytokines (IFN-gamma, IL-2), and slightly increased the production of Th2 cytokines (IL-4, IL-10) in vitro. It was observed that gastric-tumor-bearing rats administrated with MG7-scFv/SEB showed more inflammatory cell infiltration, more significant tumor inhibition, and longer survival time than those of rats treated with SEB or NS (Normal Saline). The data indicated that MG7-scFv/SEB fusion protein could specifically target gastric cancer cell, enhance the activity of T cells and induce tumor cell apoptosis to exert the antitumor effect on gastric cancer.

Role of IL-17 and TGF-β in peritoneal adhesion formation after surgical trauma

Peritoneal adhesions are fibrous tissues formed after surgery. Both cytokines and transforming growth factors (TGFs) are involved in this process. The objective of this study was to investigate the cross talk between these entities. Peritoneal drainage fluid after surgery from patients and rodent models was examined by enzyme‐linked immunosorbent assay and fluorescence‐activated cell sorter. Data showed that the concentrations of interferon (IFN)‐γ and interleukin (IL)‐17 reached their peaks 6–12 hours after surgery, whereas TGF‐β1 concentrations showed two postoperative peak time points at 2 and 72–96 hours. By neutralizing IFN‐γ, IL‐17 6–12 hours, and TGF‐β1 72–96 hours after surgery, the degree of adhesion reduced significantly. However, neutralizing TGF‐β1 2 hours after surgery did not affect adhesion formation. Furthermore, in vitro studies showed that compared with the fibroblasts that were directly stimulated with TGF‐β1, the prestimulation of IL‐17 promoted plasminogen activator inhibitor‐1 production while inhibiting tissue‐type plasminogen activator production. Moreover, additional stimulation with IFN‐γ enhanced this effect. Together, these data indicate that IL‐17 may promote adhesion formation by increasing the reaction of fibroblasts against TGF‐β1. Blocking IL‐17 might have a therapeutic potential in preventing adhesion formation after surgery.

Siglec-10 is associated with survival and natural killer cell dysfunction in hepatocellular carcinoma

BACKGROUND The interaction between Siglec-10 and its ligand, CD24, selectively represses tissue damage-caused immune responses. However, the nature of Siglec-10 and CD24 in human hepatocellular carcinoma (HCC) is still poorly defined. Hereon, the expression, function, and regulation of CD24 and Siglec-10 in HCC were investigated in the present study. METHODS Flow cytometry was performed to examine the expression of Siglec-10 in HCC tissues and adjacent non-tumor tissues of HCC patients. To further determine whether Siglec-10 expression is associated with the clinical characteristics and survival, conventional immunohistochemistry was performed in 96 HCC patients. Additionally, the role of Siglec-10 in the regulation of natural killer (NK) cell dysfunction was evaluated. Finally, CD24 expression in HCC was also assessed. RESULTS Siglec-10 was expressed most on NK cells in HCC (40.7 ± 4.5%). Compared with surrounding non-tumor tissues, tumor tissues had higher Siglec-10 expression (31.0 ± 1.7% versus 40.7 ± 4.5%, n = 10, P < 0.05), and the expression was negatively associated with patient survival. Siglec-10(+)CD56(+) NK cells exhibited reduced effector function, as shown by decreased granules and cytokine expressions compared with Siglec-10(-)CD56(+) NK cells. Moreover, the number of CD24(+)CD45(-) cells in HCC tissues was higher than that in adjacent non-tumor tissues (9.4 ± 0.9% versus 3.1 ± 0.9%, n = 15, P < 0.05). CONCLUSIONS These findings suggest that Siglec-10 is associated with decreased survival and impaired NK cell function in human HCC. This process may function via the CD24-Siglec-10 interaction, which may represent a therapeutic target in HCC patients.

The Diagnostic Performance of Stool DNA Testing for Colorectal Cancer: A Systematic Review and Meta-Analysis.

Abstract This meta-analysis was designed to evaluate the diagnostic performance of stool DNA testing for colorectal cancer (CRC) and compare the performance between single-gene and multiple-gene tests. MEDLINE, Cochrane, EMBASE databases were searched using keywords colorectal cancers, stool/fecal, sensitivity, specificity, DNA, and screening. Sensitivity analysis, quality assessments, and performance bias were performed for the included studies. Fifty-three studies were included in the analysis with a total sample size of 7524 patients. The studies were heterogeneous with regard to the genes being analyzed for fecal genetic biomarkers of CRC, as well as the laboratory methods being used for each assay. The sensitivity of the different assays ranged from 2% to 100% and the specificity ranged from 81% to 100%. The meta-analysis found that the pooled sensitivities for single- and multigene assays were 48.0% and 77.8%, respectively, while the pooled specificities were 97.0% and 92.7%. Receiver operator curves and diagnostic odds ratios showed no significant difference between both tests with regard to sensitivity or specificity. This meta-analysis revealed that using assays that evaluated multiple genes compared with single-gene assays did not increase the sensitivity or specificity of stool DNA testing in detecting CRC.