The increased ratio of Treg/Th2 in promoting metastasis of hepatocellular carcinoma.

OBJECTIVE: Liver cancer is the fifth most common cancer in the world. Research on the pathogenesis and detailed molecular mechanisms of liver cancer is very important. The immune system plays an important role in regulating the incidence and metastasis of liver cancer. MATERIALS AND METHODS: This work collected 20 blood samples from patients with clinical hepatocellular carcinoma without metastasis, 20 blood samples from patients with metastatic hepatocellular carcinoma, and 20 blood samples from healthy subjects. Flow cytometry was used to analyze the content of Treg and Th2 cells in the three groups of blood samples. Immunofluorescence was applied to analyze the relative expression of CTLA-4 and CD28 in lymphocytes of each group of blood samples. Western blot was used to analyze the T cell surface protein CTLA-4, CD28, GATA3, and FOXP3 expression in each group of blood samples. RESULTS: The expression of CD28 and GATA3 in the blood of patients with hepatocellular carcinoma without metastasis was obviously higher than that of patients with metastasis of hepatocellular carcinoma, which is contrary to the expression trend of CTLA-4 and FOXP3, and corresponds to the content ratio of Treg and Th2 cells, thus verifying the relationship between Treg/Th2 ratio and metastasis of hepatocellular carcinoma. CONCLUSIONS: In the microenvironment of liver cancer, the ratio of Treg/Th2 will increase significantly, thereby promoting the metastasis of hepatocellular carcinoma.

OBJETIVO: El cáncer de hígado es el quinto cáncer más común en el mundo. La investigación sobre la patogenia y los mecanismos moleculares detallados del cáncer de hígado es muy importante. El sistema inmunológico juega un papel importante en la regulación de la incidencia y metástasis del cáncer de hígado. MATERIAL Y MÉTODOS: Este trabajo recogió 20 muestras de sangre de pacientes con carcinoma hepatocelular clínico sin metástasis, 20 muestras de sangre de pacientes con carcinoma hepatocelular metastásico y 20 muestras de sangre de sujetos sanos. Se utilizó citometría de flujo para analizar el contenido de células Treg y Th2 en los tres grupos de muestras de sangre. Se aplicó inmunofluorescencia para analizar la expresión relativa de CTLA-4 y CD28 en linfocitos de cada grupo de muestras de sangre. Se utilizó Western blot para analizar la expresión de la proteína de superficie de células T CTLA-4, CD28, GATA3, FOXP3 en cada grupo de muestras de sangre. RESULTADOS: La expresión de CD28 y GATA3 en la sangre de pacientes con carcinoma hepatocelular sin metástasis fue obviamente mayor que la de pacientes con metástasis de carcinoma hepatocelular, lo cual es contrario a la tendencia de expresión de CTLA-4 y FOXP3, y corresponde al contenido relación de células Treg y Th2, verificando así la relación entre la relación Treg/Th2 y la metástasis del carcinoma hepatocelular. CONCLUSIONES: En el microambiente del cáncer de hígado, la proporción de Treg/Th2 aumentará significativamente, promoviendo así la metástasis del carcinoma hepatocelular.

Integrin αV Mediates the Effects of Irisin on Human Mature Adipocytes.

Introdution and Aims: The myokine irisin is critical to modulating adipocytes thermogenesis and influence whole-body metabolism. However, whether there is difference in the effects of irisin on adipocytes derived from different depots remains unknown, and the receptor of irisin on adipocytes is still unclear. In this study, we determine the browning effect of irisin on adipocytes of subcutaneous and visceral human adipose tissue and explore the possibility that integrin αV was the receptor of irisin on human adipocytes. METHODS: Human adipose-derived stem cells were isolated from human subcutaneous and visceral white adipose tissues and induced to differentiate into mature adipocytes, and the expression of UCP1 and thermogenic genes in mature adipocytes were examined with or without irisin treatment and compared between groups of different adiposity and different spots. Immunoprecipitation analysis was used to detect the interaction between irisin and integrin αV on adipocytes, and the protein expression of integrin αV in adipocytes was also compared between groups of different adiposity and anatomic position. RESULTS: Irisin treatment could increase the expression level of beige adipocyte marker protein UCP1 and specific thermogenic genes in mature adipocytes derived from subcutaneous white adipose tissue but not in visceral adipose tissue. The results of immunoprecipitation showed that irisin could be attached to integrin αV on mature adipocytes, and there was no significant difference in the gene and protein expression of integrin αV in adipocytes, either derived from subcutaneous and visceral adipose tissue, or derived from obese and normal-weight individuals. CONCLUSION: The results of the present study indicated that irisin contributed to the transformation of mature white adipocytes to beige adipocytes in human subcutaneous adipose tissue but not in visceral adipose tissue. Integrin αV may mediate the browning effects of irisin on human mature adipocytes, which could provide the potential therapeutic targets for obesity and metabolic syndrome by promoting human brown adipose tissue activity.

MKL1 regulates hepatocellular carcinoma cell proliferation, migration and apoptosis via the COMPASS complex and NF-κB signaling.

BACKGROUND: Histone modification plays essential roles in hepatocellular carcinoma (HCC) pathogenesis, but the regulatory mechanisms remain poorly understood. In this study, we aimed to analyze the roles of Megakaryoblastic leukemia 1 (MKL1) and its regulation of COMPASS (complex of proteins associated with Set1) in HCC cells. METHODS: MKL1 expression in clinical tissues and cell lines were detected by bioinformatics, qRT-PCR and western blot. MKL1 expression in HCC cells were silenced with siRNA, followed by cell proliferation evaluation via Edu staining and colony formation, migration and invasion using the Transwell system, and apoptosis by Hoechst staining. HCC cell tumorigenesis was assessed by cancer cell line-based xenograft model, combined with H&E staining and IHC assays. RESULTS: MKL1 expression was elevated in HCC cells and clinical tissues which was correlated with poor prognosis. MKL1 silencing significantly repressed proliferation, migration, invasion and colony formation but enhanced apoptosis in HepG2 and Huh-7 cells. MKL1 silencing also inhibited COMPASS components and p65 protein expression in HepG2 and Huh-7 cells. HepG2 cell tumorigenesis in nude mice was severely impaired by MKL1 knockdown, resulted into suppressed Ki67 expression and cell proliferation. CONCLUSION: MKL1 promotes HCC pathogenesis by regulating hepatic cell proliferation, migration and apoptosis via the COMPASS complex and NF-κB signaling.

The MRTF-A/B function as oncogenes in pancreatic cancer.

Despite evidence that MRTF-A/B, co-activators of serum response factor (SRF), promotes tumor cell invasion and metastasis in cancer, there are no studies describing MRTF-A/B in pancreatic cancer. To clarify involvement of MRTF-A/B expression in pancreatic cancer, we used quantitative reverse transcription-polymerase chain reaction and western blot analysis to detect MRTF-A/B in pancreatic cancer, intraductal papillary mucinous neoplasm (IPMN) and non-neoplastic pancreata. MRTF-A/B expression differs significantly between cancer and non-neoplastic tissues as well as between non-neoplastic tissues and IPMN bulk tissues. Next, we studied the roles of MRTF-A/B in vitro. Overexpression of MRTF-A/B promoted epithelial-mesenchymal transition (EMT) and generated stem cell-like cells in normal pancreatic cells. We performed quantitative reverse transcription-polymerase chain reaction to detect the level of MRTF-A/B in 19 pancreatic cancer cell lines. We found that their expression was associated with gemcitabine resistance. Like in normal pancreatic cells, MRTF-A/B also promoted EMT and promoted formation of stem cell-like cells in pancreatic cancer and they could regulate microRNA expression associated with EMT and CICs. Finally, to further demonstrate the roles of MRTF-A/B in vivo, we performed nude mouse model of s.c. xenograft and found that overexpression of MRTF-A and MRTF-B promoted pancreatic cancer growth. Elucidating the roles of MRTF-A/B will help us to further understand molecular basis of the disease and offer new gene targets for effective therapies.

Association between CTLA-4 rs231775 polymorphism and hepatocellular carcinoma susceptibility.

AIMS: Our study aimed to investigate the association of cytotoxic T-lymphocyte antigen-4 (CTLA-4) rs231775 polymorphism with hepatocellular carcinoma (HCC) susceptibility. METHODS: Genotypes distribution of the control was tested by Hardy-Weinberg Equilibrium (HWE). CTLA-4 rs231775 polymorphism was analyzed in 80 patients with HCC and 78 healthy controls by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method, and the expression level of CTLA-4 in the serum of all subjects was detected using enzyme linked immunosorbent assay (ELISA) kit. Odd ratio (OR) with 95% confidence interval (CI) were calculated by chi-squared test to determine the correlation of CTLA-4 rs231775 polymorphism and the risk of HCC. RESULTS: The genotypes frequencies of the control group were in accordance with HWE. The frequencies of genotype AA and allele A in CTLA-4 rs231775 polymorphism were significantly higher in cases than the control group (AA vs. GG: OR=2.81, P=0.043; A vs. G: OR=1.63, P=0.022). Meanwhile, the expression level of CTLA-4 was remarkably higher in cases compared with the controls. The association analysis indicated that AA genotype carriers exhibited highest level of CTLA-4 (P<0.01). CONCLUSIONS: The genotype AA and allele A of CTLA-4 rs231775 polymorphism may have negative effects on HCC by modifying the expression and functions of CTLA-4.

Lentivirus-mediated LIGHT overexpression inhibits human colorectal carcinoma cell growth in vitro and in vivo.

Human LIGHT (lymphotoxin-related inducible ligand that competes for glycoprotein D binding to herpesvirus entry mediator on T cells) is the 14th member of the tumor necrosis factor (TNF) superfamily and is therefore also known as TNFSF14. LIGHT has been proven to be a multifunctional molecule affecting cell proliferation, differentiation and a number of other biological processes, in particular, cell growth inhibition. However, the expression and molecular mechanisms of the LIGHT gene in human colorectal carcinoma cells remain largely unclear. In the present study, the LIGHT gene was overexpressed using a lentiviral expression vector in HCT116 human colorectal carcinoma cells in vitro and in vivo, in order to explore the mechanism by which the LIGHT gene inhibits cell growth and suppresses tumor formation. The results showed that the recombinant lentivirus with LIGHT overexpression inhibited the proliferative capacity of the HCT116 cells and significantly decreased the xenografted tumor volumes in nude mice. Furthermore, LIGHT treatment effectively initiated increased caspase-3 and decreased Bcl-2 activities in the HCT116 cells. This study provides a basis for the improved understanding of the role and molecular mechanisms of the LIGHT gene in human colorectal carcinoma cells and may facilitate further functional studies of LIGHT.