Antibodies, Neutralizing ; Apoptosis ; Hep G2 Cells ; Humans ; Receptors, Tumor Necrosis Factor, Member 6b ; immunology ; metabolism

Objective: To investigate the effect of microRNA (miR-148b) targeting decoy receptor 3 (DcR3) on macrophage polarization in sepsis. Methods: Experimental study. From December 2019 to December 2022, serum microRNA expression was detected in 3 patients with sepsis and 3 healthy controls in the clinical laboratory of Songjiang Hospital Affiliated to Shanghai Jiao Tong University School of Medicine. Phorbol 12-myristate 13-acetate (PMA) was used to induce the differentiation of human acute monocytic leukemia cells THP-1 into macrophages, and then lipopolysaccharide (LPS) was added to stimulate the establishment of a sepsis cell model, and the expression changes of miR-148b and DcR3 were detected by RT-PCR and Western blot. Overexpression of DcR3 was used to detect the expression levels of TNF-α, CD163 and IL-10 in macrophages stimulated by LPS (100 ng/ml). Overexpression of miR-148b was used to observe the changes of molecular markers of macrophage polarization. The targeting regulation effect of miR-148b on DcR3 was determined by dual-luciferase reporter assay. t test was used to analyze whether there were statistical differences among the groups. Results: The expression of miR-148b was down-regulated (P<0.05) and the expression of DcR3 was up-regulated (P<0.01) in THP-1 macrophages stimulated by LPS. Overexpression of DcR3 inhibited the expression of TNF-α (P<0.05) and promoted the expression of CD163 (P<0.01) and IL-10 (P<0.01). When miR-148b mimics was added, the opposite effect was observed. The dual-luciferase reporter assay confirmed that miR-148b targets and binds to DcR3, inhibiting its transcription and expression. The results of flow cytometry showed that DcR3 could reverse the promoting effect of miR-148b on the CD86/CD163 ratio of macrophages (P<0.05). Conclusion: miR-148b inhibits the expression of DcR3, thereby inhibiting M2 polarization in LPS-stimulated macrophage cells.
Humans ; Interleukin-10 ; Lipopolysaccharides/pharmacology* ; Macrophages ; MicroRNAs/genetics* ; Receptors, Tumor Necrosis Factor, Member 6b/metabolism* ; Tumor Necrosis Factor-alpha

Objective: To investigate the effect of microRNA (miR-148b) targeting decoy receptor 3 (DcR3) on macrophage polarization in sepsis. Methods: Experimental study. From December 2019 to December 2022, serum microRNA expression was detected in 3 patients with sepsis and 3 healthy controls in the clinical laboratory of Songjiang Hospital Affiliated to Shanghai Jiao Tong University School of Medicine. Phorbol 12-myristate 13-acetate (PMA) was used to induce the differentiation of human acute monocytic leukemia cells THP-1 into macrophages, and then lipopolysaccharide (LPS) was added to stimulate the establishment of a sepsis cell model, and the expression changes of miR-148b and DcR3 were detected by RT-PCR and Western blot. Overexpression of DcR3 was used to detect the expression levels of TNF-α, CD163 and IL-10 in macrophages stimulated by LPS (100 ng/ml). Overexpression of miR-148b was used to observe the changes of molecular markers of macrophage polarization. The targeting regulation effect of miR-148b on DcR3 was determined by dual-luciferase reporter assay. t test was used to analyze whether there were statistical differences among the groups. Results: The expression of miR-148b was down-regulated (P<0.05) and the expression of DcR3 was up-regulated (P<0.01) in THP-1 macrophages stimulated by LPS. Overexpression of DcR3 inhibited the expression of TNF-α (P<0.05) and promoted the expression of CD163 (P<0.01) and IL-10 (P<0.01). When miR-148b mimics was added, the opposite effect was observed. The dual-luciferase reporter assay confirmed that miR-148b targets and binds to DcR3, inhibiting its transcription and expression. The results of flow cytometry showed that DcR3 could reverse the promoting effect of miR-148b on the CD86/CD163 ratio of macrophages (P<0.05). Conclusion: miR-148b inhibits the expression of DcR3, thereby inhibiting M2 polarization in LPS-stimulated macrophage cells.
Humans ; Interleukin-10 ; Lipopolysaccharides/pharmacology* ; Macrophages ; MicroRNAs/genetics* ; Receptors, Tumor Necrosis Factor, Member 6b/metabolism* ; Tumor Necrosis Factor-alpha

OBJECTIVETo evaluate the value of serum TR(6) for the diagnosis and TNM classification in patients with gastric carcinoma.

METHODSSerum TR(6) levels were measured using ELISA method in 31 gastric cancer patients, 19 patients with nonmalignant conditions and 29 healthy individuals. TR(6) expression in tumor mass was studied with immunohistochemistry. TR(6) gene copy number in tumor tissues was evaluated by real time PCR.

RESULTSNinety-seven point nine percent (47 of 48 cases) of healthy individuals and patients with nonmalignant conditions were serum TR(6)-negative. In contrast, 71% (22 of 31 cases) of gastric cancer patients were serum TR(6)-positive. Serum TR(6) positiveness was closely correlated with tumor differentiation status and TNM classification. TR(6) gene amplification did not occur in gastric carcinoma.

CONCLUSIONSSerum TR(6) levels were correlated significantly with TNM stage and histopathological type of tumor. This can help to determine the pre-operative TNM classification and to choose the optimal extent of lymph node dissection for gastric cancer.

Humans ; Lymphatic Metastasis ; Membrane Glycoproteins ; blood ; Neoplasm Staging ; Receptors, Cell Surface ; blood ; Receptors, Tumor Necrosis Factor ; Receptors, Tumor Necrosis Factor, Member 6b ; Stomach Neoplasms ; blood ; pathology

OBJECTIVETo investigate the association of serum levels of decoy receptor 3(DcR3) protein and the clinicopathologic features of bladder transitional cell carcinoma.

METHODSEnzyme-linked immunosorbent assay was used to examine the serum levels of DcR3 in patients with bladder transitional cell carcinoma for analysis of its association with the patients' age, gender, clinical stages and pathological classification.

RESULTSThe patients with bladder transitional cell carcinoma showed a significantly elevated serum level of DcR3 (183.43 ∓78.45 pg/m1) compared with the normal level (116.65∓97.43 pg/m1, P<0.05). The serum level of DcR3 in the patients showed close correlations with the TNM stage and pathological classification of the tumor (P<0.05) but not with the patients' age or gender (P>0.05).

CONCLUSIONSIn patients with bladder transitional cell carcinoma, a high serum level of DcR3 suggests a higher malignancy of the tumor.

Carcinoma, Transitional Cell ; blood ; Enzyme-Linked Immunosorbent Assay ; Humans ; Receptors, Tumor Necrosis Factor, Member 6b ; blood ; Urinary Bladder Neoplasms ; blood

Adult ; Aged ; Apoptosis ; Female ; Humans ; Laryngeal Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Receptors, Tumor Necrosis Factor, Member 6b ; metabolism

Adult ; Aged ; Aged, 80 and over ; Apoptosis ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Female ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Receptors, Tumor Necrosis Factor, Member 6b ; metabolism

OBJECTIVE: To investigate the expression of EMS1 and DcR3 in laryngeal carcinoma and analyze the relation of EMS1 and DcR3. METHOD: The expression of EMS1 and DcR3 protein in 41 laryngeal carcinoma fresh samples and 41 para-carcinoma tissues (to cutting margin > 0.5 cm) were measured by flow cytometry, and 15 normal laryngeal mucosa samples were also studied as controls. RESULT: (1) The quantitative and qualitative expression of EMS1 and DcR3 protein in laryngeal carcinoma tissues was obviously higher than those in para-carcinoma and in normal laryngeal mucosa tissues respectively (P < 0.05). There was no significant difference between the expression of para-carcinoma and normal laryngeal mucosa tissues. (2) In laryngeal carcinoma, the expression of EMS1 and DcR3 protein was independent of patients' clinical classification, tumor size, smoking history, patients' age and sex but associated with tumor metastasis, pathological grade and clinical stage. (3) In laryngeal carcinoma, the expression of EMS1 was positively correlated with that of DcR3. CONCLUSION EMS1 was positively related to DcR3, which might play an important role in the carcinogenesis and development of laryngeal carcinoma by synergic effect.
Adult ; Aged ; Cortactin ; metabolism ; Female ; Flow Cytometry ; Humans ; Laryngeal Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Receptors, Tumor Necrosis Factor, Member 6b ; metabolism

OBJECTIVE: To investigate the expression of DcR3 in laryngeal carcinoma and analyze the relation between DcR3 and clinical factors. METHOD: The expression of DcR3 protein in 41 laryngeal carcinoma tissues and 41 para-carcinoma tissues (to cutting margin > 0.5 cm) were measured by Flow Cytometer(Epics-XL II), 15 normal laryngeal mucosa tissues were served as controls. RESULT: (1) The quantitative and qualitative expression of DcR3 protein in laryngeal carcinoma tissues was obviously higher than those in para-carcinoma and normal laryngeal mucosa tissues, respectively (P < 0.05). There was no significant difference for the DcR3 protein expression between para-carcinoma and normal laryngeal mucosa tissues. (2) In laryngeal carcinoma, the expression of DcR3 protein was not significantly related to clinical classification, tumor size, smoking history, patients' age and sex but related to metastasis, pathological grade and clinical stage. CONCLUSION The high level of DcR3 expression may contribute to the carcinogenesis and development of laryngeal carcinoma. So it can be an important index for judging the differentiation, infiltration, metastasis and staging of laryngeal carcinoma.
Adult ; Aged ; Carcinoma ; metabolism ; pathology ; Female ; Humans ; Laryngeal Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Prognosis ; Receptors, Tumor Necrosis Factor, Member 6b ; metabolism

BACKGROUNDDecoy receptor 3 (DcR3) binds to Fas ligand (FasL) and inhibits FasL-induced apoptosis. The receptor is overexpressed in hepatocellular carcinoma (HCC), and it is associated with the growth and metastatic spread of tumors. DcR3 holds promises as a new target for the treatment of HCC, but little is known regarding the molecular mechanisms underlying the oncogenic properties of DcR3. The present work, therefore, examined the role of DcR3 in regulating the growth and invasive property of liver cancer cell HepG2.

METHODSHepG2 cells were stably transfected with lentivirus-based short hairpin RNA vector targeting DcR3. After the knockdown of DcR3 was confirmed, cell proliferation, clone formation, ability of migrating across transwell membrane, and wound healing were assessed in vitro. Matrix metalloproteinase-9 (MMP 9) and vascular epithelial growth factor (VEGF)-C and D expressions of the DcR3 knockdown were also studied. Comparisons between multiple groups were done using one-way analysis of variance (ANOVA), while pairwise comparisons were performed using Student's t test. P< 0.05 was regarded statistically significant.

RESULTSDcR3 was overexpressed in HepG2 compared to other HCC cell lines and normal hepatocyte Lo-2. Stable knockdown of DcR3 slowed down the growth of HepG2 (P < 0.05) and reduced the number of clones formed by 50% compared to those without DcR3 knockdown (P < 0.05). The knockdown also reduced the migration of HepG2 across transwell matrix membrane by five folds compared to the control (P < 0.05) and suppressed the closure of scratch wound (P < 0.05). In addition, the messenger RNA levels of MMP 9, VEGF-C, and VEGF-D were significantly suppressed by DcR3 knockdown by 90% when compared with the mock control (P < 0.05).

CONCLUSIONSLoss of DcR3 impaired the growth and invasive property of HCC cell line of HepG2. Targeting DcR3 may be a potential therapeutic approach for the treatment of HCC.

Analysis of Variance ; Cell Movement ; genetics ; physiology ; Cell Proliferation ; genetics ; physiology ; Hep G2 Cells ; Humans ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; RNA, Small Interfering ; genetics ; Receptors, Tumor Necrosis Factor, Member 6b ; genetics ; metabolism ; Vascular Endothelial Growth Factor A ; genetics ; metabolism