Sex-Biased Molecular Signature for Overall Survival of Liver Cancer Patients

Sex/gender disparity has been shown in the incidence and prognosis of many types of diseases, probably due to differences in genes, physiological conditions such as hormones, and lifestyle between the sexes. The mortality and survival rates of many cancers, especially liver cancer, differ between men and women. Due to the pronounced sex/gender disparity, considering sex/ gender may be necessary for the diagnosis and treatment of liver cancer. By analyzing research articles through a PubMed literature search, the present review identified 12 genes which showed practical relevance to cancer and sex disparities. Among the 12 sex-specific genes, 7 genes (BAP1, CTNNB1, FOXA1, GSTO1, GSTP1, IL6, and SRPK1) showed sex-biased function in liver cancer. Here we summarized previous findings of cancer molecular signature including our own analysis, and showed that sexbiased molecular signature CTNNB1High , IL6High , RHOAHigh and GLIPR1Low may serve as a female-specific index for prediction and evaluation of OS in liver cancer patients. This review suggests a potential implication of sex-biased molecular signature in liver cancer, providing a useful information on diagnosis and prediction of disease progression based on gender.

Combinatorial Antitumor Activity of Oxaliplatin with Epigenetic Modifying Agents, 5-Aza-CdR and FK228, in Human Gastric Cancer Cells

Epigenetic silencing is considered to be a major mechanism for loss of activity in tumor suppressors. Reversal of epigenetic silencing by using inhibitors of DNA methyltransferase (DNMT) or histone deacetylases (HDACs) such as 5-Aza-CdR and FK228 has shown to enhance cytotoxic activities of several anticancer agents. This study aims to assess the combinatorial effects of gene-silencing reversal agents (5-Aza-CdR and FK228) and oxaliplatin in gastric cancer cells, i.e., Epstein-Barr virus (EBV)-negative SNU-638 and EBV-positive SNU-719 cells. The doublet combinatorial treatment of 5-Aza-CdR and FK228 exhibited synergistic effects in both cell lines, and this was further corroborated by Zta expression induction in SNU-719 cells. Three drug combinations as 5-Aza-CdR/FK228 followed by oxaliplatin, however, resulted in antagonistic effects in both cell lines. Simultaneous treatment with FK228 and oxaliplatin induced synergistic and additive effects in SNU-638 and SNU-719 cells, respectively. Three drug combinations as 5-Aza-CdR prior to FK228/oxaliplatin, however, again resulted in antagonistic effects in both cell lines. This work demonstrated that efficacy of doublet synergistic combination using DNMT or HDACs inhibitors can be compromised by adding the third drug in pre- or post-treatment approach in gastric cancer cells. This implies that the development of clinical trial protocols for triplet combinations using gene-silencing reversal agents should be carefully evaluated in light of their potential antagonistic effects.

Regulation of microRNA-7-5p and LRP6 by Epstein-Barr Virus-Encoded RNAs in Burkitt's Lymphoma Cell Line Akata

Epstein-Barr virus (EBV)-encoded small non-coding RNAs (EBERs) are abundantly expressed in various EBV-associated malignancies, and play critical roles in cell proliferation, tumorigenesis, and apoptosis resistance. However, the mechanism how EBERs regulate cell function awaits further clarification. In this study, we investigated the effect of EBERs on the expression of cellular microRNA (miRNA) and mRNA expression. To test the effect of EBERs while unaffected by other EBV genes, we used EBERs-deleted recombinant EBV infected Burkitt's lymphoma cell line (Akata(+)EBERs(-)) as well as EBV-infected (Akata(+)) and EBV uninfected (Akata(-)) cell lines. They all have the same genetic backgrounds. First, 15 different cellular miRNAs which have reverse complementary sequences to EBERs and have reported targets were selected by bioinformatics analysis. When RT-PCR was carried out for the 16 miRNAs using RNAs from Akata(+), Akata(-), and Akata(+)EBERs(-) cells, hsa-miR-7-5p was the only one showing down-regulated expression in Akata(+) than in Akata(-) and Akata(+)EBERs(-) cells. Bioinformatics and mRNA microarray analyses for Akata(+), Akata(-), and Akata(+)EBERs(-) cell lines were then carried out to predict putative targets of hsa-miR-7-5p. Among the 6 predicted targets of hsa-miR-7-5p, only low density lipoprotein receptor-related protein 6 (LRP6) was up-regulated in EBERs-expressing cells when tested by RT-PCR and Western blot. However, luciferase reporter assay showed that the 3'-UTR of LRP6 was not directly targeted by hsa-miR-7-5p. Our data suggest that both hsa-miR-7-5p and LRP6 are regulated by EBERs in Akata cells, and these genes may partly mediate the tumorigenic function of EBERs in Burkitt's lymphoma.

Inhibition of Transfer Infection of Epstein-Barr Virus to Epithelial Cells by Integrin beta6 siRNA

Epstein-Barr virus (EBV) establishes a latent infection in greater than 90% of the world's adult population and associates with various tumors. EBV primarily infects epithelial cells and B cell in vivo. Mechanism of EBV infection in B cells is known to involve binding of EBV glycoprotein gp350 to CD21 on B cell surface. Epithelial cells are infected with EBV even though most of epithelial cells do not express CD21. Recently, integrin alphavbeta5, alphavbeta6 and alphavbeta8 on epithelial cells were reported to facilitate EBV infection by interacting with gHgL complex. We examined the expression profile of integrins known to be expressed on epithelial cells. Integrin alphavbeta5 and alphavbeta6, but not alphavbeta8 were detected in a gastric epithelial cell line, AGS. We then tested whether siRNAs specific to beta6 can inhibit EBV infection of epithelial cells. One among the four tested siRNAs significantly reduced beta6 expression and suppressed transfer infection of EBV to AGS cells. Our data suggest that siRNAs to integrins might be useful to control EBV infection to epithelial cells.

Repeated Gene Transfection Impairs the Engraftment of Transplanted Porcine Neonatal Pancreatic Cells

BACKGROUND: Previously, we reported that neonatal porcine pancreatic cells transfected with hepatocyte growth factor (HGF) gene in an Epstein-Barr virus (EBV)-based plasmid (pEBVHGF) showed improved proliferation and differentiation compared to those of the control. In this study, we examined if pancreatic cells transfected repeatedly with pEBVHGF can be successfully grafted to control blood glucose in a diabetes mouse model. METHODS: Neonatal porcine pancreatic cells were cultured as a monolayer and were transfected with pEBVHGF every other day for a total of three transfections. The transfected pancreatic cells were re-aggregated and transplanted into kidney capsules of diabetic nude mice or normal nude mice. Blood glucose level and body weight were measured every other day after transplantation. The engraftment of the transplanted cells and differentiation into beta cells were assessed using immunohistochemistry. RESULTS: Re-aggregation of the pancreatic cells before transplantation improved engraftment of the cells and facilitated neovascularization of the graft. Right before transplantation, pancreatic cells that were transfected with pEBVHGF and then re-aggregated showed ductal cell marker expression. However, ductal cells disappeared and the cells underwent fibrosis in a diabetes mouse model two to five weeks after transplantation; these mice also did not show controlled blood glucose levels. Furthermore, pancreatic cells transplanted into nude mice with normal blood glucose showed poor graft survival regardless of the type of transfected plasmid (pCEP4, pHGF, or pEBVHGF). CONCLUSION: For clinical application of transfected neonatal porcine pancreatic cells, further studies are required to develop methods of overcoming the damage for the cells caused by repeated transfection and to re-aggregate them into islet-like structures.

The role of promoter methylation in Epstein-Barr virus (EBV) microRNA expression in EBV-infected B cell lines

Epstein-Barr virus (EBV) microRNAs (miRNAs) are expressed in EBV-associated tumors and cell lines, but the regulation mechanism of their expression is unclear yet. We investigated whether the expression of EBV miRNAs is epigenetically regulated in EBV-infected B cell lines. The expression of BART miRNAs was inversely related with the methylation level of the BART promoter at both steady-state and following 5-aza-2'-deoxycytidine treatment of the cells. The expression of BHRF1 miRNAs also became detectable with the demethylation of Cp/Wp in latency I EBV-infected cell lines. Furthermore, in vitro methylation of the BART and Cp promoters reduced the promoter-driven transactivation. In contrast, tricostatin A had little effect on the expression of EBV miRNA expression as well as on the BART and Cp/Wp promoters. Our results suggest that promoter methylation, but not histone acetylation, plays a role in regulation of the EBV miRNA expression in EBV-infected B cell lines.

Epstein-Barr Virus Associated Lymphoproliferative Lesion Presenting as a Hydroa Vacciniforme-like Eruption / 대한피부과학회지

BACKGROUND: There have been several reports of patients with a severe hydroa vacciniforme (HV)-like eruption from Asia and Latin America. The cutaneous lesions are present in both sun-exposed and nonexposed areas in these patients unlike true HV. Several patients have died of malignant hematologic malignancies. The latent Epstein-Barr virus (EBV) infection has been detected in the skin lesions of the patients. OBJECTIVE: To describe clinical, histological, immunohistochemical, and molecular pathological features of the patients with EBV associated lymphoproliferative lesion presenting as a HV-like eruption. METHODS: The clinical, histological, and immunohistochemical features of 16 patients were reviewed. The presence of T-cell receptor (TCR)-gamma gene rearrangement was investigated using polymerase chain reaction (PCR) technique. Photoprovocation by repetitive UVA exposure was performed in five patients. In situ hybridization was performed to detect mRNA for EBV in the lesional skin biopsy specimen, lymph node biopsy specimen, mucosal biopsy specimen of stomach, and the skin biopsy specimen of photo-provoked site. PCR was performed to detect DNA for EBV in the skin biopsy specimens of 6 patients and peripheral mononuclear cells of 2 patients. RESULTS: The severity of the skin lesion and the clinical course varied among the patients. Skin biopsy specimens obtained from a papule or a vesicle showed perivascular and periadnexal infiltrate of lymphoid cells with T-cell phenotype. However, clonal TCR-gamma gene rearrangement was not detected in all 8 patients. Papules or vesicles were induced by repetitive UVA exposure in 5 patients. A latent EBV infection was demonstrated in all the tested samples, such as lesional skin, lymph node, gastric mucosa, peripheral blood mononuclear cells, and the photo-provoked lesion. CONCLUSION: EBV associated lymphoproliferative lesion presenting as a HV-like eruption is a novel disease that is not related to classic HV. Repetitive irradiation of UVA can induce the skin lesion in some patients with EBV associated lymphoproliferative lesion presenting as a HV-like eruption.

Highly Efficient Gene Expression in Rabbit Synoviocytes Using EBV-Based Plasmid

BACKGROUND: Rheumatoid arthritis (RA) is an autoimmune disorder characterized by chronic synovial inflammation which leads to joint destruction. Gene therapy of RA targets the players of inflammation or articular destruction. However, viral vectors have safety problems and side effects, while non-viral vectors suffer from inefficient gene transfer and fast loss of gene expression. To overcome the limits of non-vial vectors, an EBV-based plasmid which is known to exert prolonged high level gene expression can be used. METHODS: pEBVGFP, pEBVIL-10, and pEBVvIL-10 were constructed by cloning GFP, IL-10, and vIL-10 genes into an EBV-based plasmid, respectively. The pGFP was used as a control plasmid. Each constructs were lipofected into HIG-82 rabbit synoviocytes. The expression of GFP was monitored by FACS and confocal microscopy. IL-10 and vIL-10 expressions were measured by ELISA. RESULTS: GFP expression 2 days after transfection was achieved in 33.2% of cells. GFP-expressing cells transfected with pGFP decreased rapidly from 4 days after transfection and disappeared completely by 11 days. Cells transfected with pEBVGFP began to decrease slowly from 4 days. But GFP expression was detected for over 35 days. In addition, HIG-82 cells transfected with pEBVIL-10 (44.6+/-1.5 ng/ml) or pEBVvIL-10 (51.0+/-5.7 ng/ml) secreted these cytokines at high levels. High level cytokine production by hygromycin selection was maintained at least for up to 26 days after transfection. CONCLUSION: These results suggest that the EBV-based plasmid has a potential to improve non-viral gene transfer system and may be applicable to treat RA without the drawbacks of viral vectors.

Lack of Association between Epstein-Barr Virus and Epithelial Malignancies Developed after Kidney Transplantation / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: Organ transplant recipients are at high risk of developing malignancies due to immunosuppressive regimens. Unlike post-transplant lymphoproliferative diseases (PTLDs), where Epstein-Barr virus (EBV) plays an etiological role, there are conflicting data regarding the association of EBV with post-transplant epithelial malignancies. In order to clarify the role of EBV in carcinomas that develop after solid-organ transplantation, the presence of EBV infection in the carcinomas of post-kidney transplant patients was examined. MATERIALS AND METHODS: The presence of EBV infection in skin carcinoma (PTSC), gastric carcinoma (PTGC) and urothelial carcinoma (PTUC), which developed in the patients under an immune suppression regime following kidney transplantation, was examined. Tumors from the patients without organ transplantation were also used as a comparison in the study. The study group included five nasopharyngeal carcinomas (NPCs), one Hodgkin's disease (HD), one B-cell non-Hodgkin's malignant lymphoma (NHL) and one hypopharynx (HPC) tumor. RESULTS: Immunofluorescence assay and Western blot analysis, using sera from the same patients, confirmed that all of the tested patients were previously infected with EBV. From in situ hybridization, no EBER positive cells were detected in any of the tumor tissues obtained from the three kidney transplant recipients (PTSC, PTGC and PTUC) or in the NHL and HPC tissues. In contrast, all five of the NPC and HD tissues showed strong EBER positivity. CONCLUSION: These results suggest that there is a strong association of EBV with NPC and HD as previously reported, while no such strong association of EBV was found with epithelial malignancies that developed after kidney transplantation.

Detection Rates of Epstein-Barr Virus from the Chronic Tonsillitis and Its Typing / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Epstein-Barr virus (EBV) is a DNA virus and a type of Herpes virus. Two different types of EBV exist based on the DNA sequence divergence of the EBV genome, and several differences exist between the two types. There are no reports about EBV detection rates and typings associated in chronic tonsillitis, despite the known fact that tonsil is one of the most common sites EBV resides. The aims of this study were to investigate the prevalence of EBV infection in tonsils of children and adults as well as to compare the detection rates and the distribution of types of EBV in tonsillitis patients with normal controls. MATERIALS AND METHODS: Palatine tonsil tissues were obtained from seventy adults (20 normal controls and 50 chronic tonsillitis) and one hundred children. Using polymerase chain reaction (PCR), EBV genome was detected and typing was performed. We confirmed EBV by Southern blot hybridization and in situ hybridization. RESULTS: The detection rates of EBV in chronic tonsillitis of adults were higher than those in children with idiopathic tonsillar hypertrophy(ITH) and normal controls. The detection rates of type 1 EBV showed no differences among the four groups. The detection rates of type 2 EBV in ITH and chronic tonsillitis of adults and children were higher than those in the normal controls. CONCLUSION: Our results suggest that tonsillitis is caused by EBV and that the type 2 EBV plays a more important role in tonsillitis.

The Regulation of TRAF Expression by TNF-alpha in Rheumatoid Synoviocytes / 대한면역학회지

No abstract available.

The Clinical Significance of IgG Antibodies to Native Human Type II Collagen in Patients with Rheumatoid Arthritis / 대한면역학회지

OBJECTIVE: To determine the clinical significance of IgG antibodies to native type II collagen (IgG nCII) in patients with rheumatoid arthritis (RA). METHODS: IgG antibodies to native type II collagen (anti-HnCII) were measured in 287 patients with RA, 34 patients with osteoarthritis (OA) and 50 normal controls by improved ELISA using avidin-biotin system and 100% normal goat serum. The clinical and laboratory variables were investigated in patients with RA at the time of sampling. RESULTS: The titers of anti-HnCII were higher in RA than OA and normal control (median value 5.2 in RA, 3.0 in OA, 1.7 a.u in normal) (p3 year). There was no differences in age, sex, erosions on X-ray, mean steroid dose, use of DMARD, extraarticular manifetations such as lung involvement, rheumatoid nodule, Sjogren's syndrome between groups with anti-HnCII positive and negative. However, deformity was more frequently found in anti-HnCII negative (p<0.05). Among the 83 patients measured anti-HnCII serially (mean interval; 15+/-6 month), 11 of 24 patients with anti- HnCII positve (45.8%) were converted to negative and 19 of 59 patients with anti- HnCII negative (32.2%) were converted to positive. The levels of CRP decreased in groups converted from positive to negative (p<0.05) and vice versa (p<0.05). CONCLUSION: These results suggest that the levels of IgG HnCII fluctuate with positive correlation with acute phase reactants and may reflect the inflammatory activity of RA.