Expression Of Zinc Finger Protein Research Articles

Studies on the expression and biological functions of ZIC5 in hepatocellular carcinoma

Purpose: To study the expression of zinc finger protein of the cerebellum 5 (ZIC5) and its biological functions in hepatocellular carcinoma (HCC). Methods: Sixty-five patients undergoing HCC surgery were selected. Expression of ZIC5 in HCC and para-carcinoma tissue was examined by quantitative real time-polymerase chain reaction (qRT-PCR) and western blotting. The relationship between ZIC5 expression and clinicopathological features, postoperative survival rate, and prognosis of liver cancer patients was analyzed by t-test, Kaplan-Meier method, and Cox regression analysis, respectively. The effects of ZIC5 silencing on Huh-7 cell proliferation, migration, invasion, and apoptosis were assessed using Cell Counting Kit-8 (CCK-8), wound healing assay, Transwell assay, and flow cytometry, respectively. Results: ZIC5 expression in liver cancer tissue was significantly higher than in the para-carcinoma tissue and was significantly correlated with TNM stage and differentiation degree (p < 0.001). The overall survival rate of patients with high ZIC5 expression level was significantly lower than that of patients with low ZIC5 expression (p < 0.01). ZIC5 expression, TNM stage, and differentiation degree were independent prognostic factors. ZIC5 silencing significantly inhibited the proliferative, migratory, invasive, and anti-apoptotic capacity of Huh-7 cells (p < 0.01). Conclusion: ZIC5 is highly expressed in HCC, and this can promote liver cancer cell proliferation, migration, and invasion.

Expression of zinc finger protein 217 in non-small cell lung cancer and its clinical significance

Objective To explore the expression of zinc finger protein 217 (ZNF217) in non-small cell lung cancer (NSCLC) and its correlation with prognosis of patients. Methods A total of 120 NSCLC patients in Chongqing Three Gorges Central Hospital from January 2012 to October 2013 were selected. Immunohistochemical method was used to test the expression of ZNF217 in NSCLC tissues and paracancerous tissues. The correlation of ZNF217 expression with patient's clinicopathological features was analyzed. At the same time, the Kaplan-Meier method and Cox regression model multiple factor analysis method were used to explore the factors affecting the prognosis of patients after NSCLC radical operations. Results ZNF217 mainly existed in cell nucleus of NSCLC. The positive expression rate of ZNF217 in the cancer tissues was higher than that in the paracancerous tissues [52.5% (63/120) vs. 20.1% (25/120), χ2= 25.909, P < 0.05]. The positive expression rate of ZNF217 increased with the increase of tumor T stage (χ 2= 7.333, P= 0.026), N stage (χ 2= 7.782, P= 0.020) and TNM stage (χ 2= 11.557, P= 0.003). The overall survival (P= 0.007) and progression-free survival (P= 0.004) of patients with positive ZNF217 were poorer than those of patients with negative ZNF217. Cox multiple factor analysis showed that ZNF217 was an independent risk factor affecting the prognosis of NSCLC. Conclusion ZNF217 is an independent risk factor affecting the prognosis of NSCLC, and it may be a potential target for accurate treatment of NSCLC. Key words: Carcinoma, non-small-cell lung; Zinc finger protein 217; Prognosis

Expression of ZNF207 in hepatocellular carcinoma and its significance

To investigate the expression of zinc finger protein 207 (ZNF207) in hepatocellular carcinoma (HCC) tissues, and analyze the correlation of ZNF207 with clinicopathological factors and HCC patients' survival. Methods: Real-time PCR was used to detect ZNF207 mRNA expression in 10 paired fresh HCC and adjacent non-tumor liver tissue samples. Immunohistochemical (IHC) analysis was used to detect ZNF207 protein expression in 135 cases of randomly selected paraffin-embedded HCC tissues. The correlation of ZNF207 expression with clinicopathological factors and survival of HCC was analyzed. Results: The ZNF207 mRNA expression level in HCC was significantly higher than that in the adjacent non-tumor liver tissue (P<0.01). IHC results showed that ZNF207 protein level was elevated in HCC tissues and ZNF207 expression was correlated with cirrhosis, nodule number, tumor capsule, vascular invasion, and TNM stage (P<0.05). Survival analysis showed that patients with high ZNF207 expression had poorer overall survival (OS) and disease-free survival (DFS) than those with the low ZNF207 expression (P<0.01), and ZNF207 was an independent risk factor for OS and DFS of HCC (P<0.05). Conclusion: ZNF207 expression is elevated in HCC and associated with adverse clinicopathological factors, indicating poor prognosis for HCC.

ZFP91 zinc finger protein expression pattern in normal tissues and cancers.

Zinc finger protein 91 (ZFP91) gene has been recently acknowledged to possess oncogenic properties. To date, its expression has been examined only in a handful of human organs and cancer types. The aim of the present study was to characterize, for the first time, the ZFP91 expression pattern in a range of human tissues and cancer types. ZFP91 mRNA expression was examined using Cancer Survey cDNA sets. Utilized cDNA samples represented 15 human organs and 17 cancer types. ZFP91 mRNA expression was the highest in the testes and lymph nodes. It was downregulated in testis cancer, lymphoma and thyroid cancer, and upregulated in prostate cancer. Among the analyzed cancer types, ZFP91 expression was markedly elevated in sarcomas and melanoma. On a protein level, a large-scale reverse phase protein array was employed providing samples from 11 organ types and from cancers derived from these organs. ZFP91 protein expression was revealed to be generally stable across the tested samples and was only moderately elevated in breast, ovarian and pancreatic cancers. To the best of our knowledge, this is the first study to thoroughly analyze the ZFP91 expression pattern in human tissues and cancers. The obtained results provide the foundation for further work aiming to reveal its full biological significance.

Zinc Finger Protein 521, Negatively Regulated by MicroRNA-204-5p, Promotes Proliferation, Motility and Invasion of Gastric Cancer Cells.

Objective:This study aims to investigate the expression, role, and detailed mechanism of microRNA-204-5p and zinc finger protein 521 in gastric cancer.Methods:Immunohistochemistry was adopted to detect the expressions of zinc finger protein 521 in 82 cases of gastric cancer tissues. Western blot was used to detect the expressions of zinc finger protein 521 in gastric cancer cells and adjacent cells. Moreover, the correlation between zinc finger protein 521 and the prognosis of patients were also evaluated. Cell Counting Kit 8 assay and colony formation assay were performed to figure out the impact of zinc finger protein 521 on the proliferation of gastric cancer cells. By conducting flow cytometry, the effect of zinc finger protein 521 on the apoptosis of gastric cancer cells was determined. The scratch wound healing assay and transwell invasion assay were carried out to determine the effect of zinc finger protein 521 on regulating the motility and invasion of gastric cancer cells. Ultimately, the targeting relationship and interaction between microRNA-204-5p and zinc finger protein 521 were verified by real-time polymerase chain reaction, Western blot, and dual luciferase reporter gene assay.Results:Compared with adjacent cells, zinc finger protein 521 was highly expressed in gastric cancer cells, which was related to TNM stage (P = .0388), tumor size (P = .0168), and local lymph node metastasis (P = .0024). Overexpressed zinc finger protein 521 can promote the proliferation, migration, and invasion of gastric cancer cells and inhibit the apoptosis. Zinc finger protein 521 is a target gene of microRNA-106-5p, and there was a negative correlation between the expression of zinc finger protein 521 and microRNA-204-5p.Conclusion:Zinc finger protein 521 can arrest the apoptosis and enhance the proliferation, migration, and invasion of gastric cancer cells via regulating microRNA-204-5p. Our study may provide novel clues for the treatment of patients with gastric cancer.

Sex-Specific Regulation of Fear Memory by Targeted Epigenetic Editing of Cdk5

BackgroundSex differences in the expression and prevalence of trauma- and stress-related disorders have led to a growing interest in the sex-specific molecular and epigenetic mechanisms underlying these diseases. Cyclin-dependent kinase 5 (CDK5) is known to underlie both fear memory and stress behavior in male mice. Given our recent finding that targeted histone acetylation of Cdk5 regulates stress responsivity in male mice, we hypothesized that such a mechanism may be functionally relevant in female mice as well. MethodsWe applied epigenetic editing of Cdk5 in the hippocampus and examined the regulation of fear memory retrieval in male and female mice. Viral expression of zinc finger proteins targeting histone acetylation to the Cdk5 promoter was paired with a quantification of learning and memory of contextual fear conditioning, expression of CDK5, and enrichment of histone modifications of the Cdk5 gene. ResultsWe found that male mice exhibit stronger long-term memory retrieval than do female mice, and this finding was associated with male-specific epigenetic activation of hippocampal Cdk5 expression. Sex differences in behavior and epigenetic regulation of Cdk5 occurred after long-term, but not short-term, fear memory retrieval. Finally, targeted histone acetylation of hippocampal Cdk5 promoter attenuated fear memory retrieval and increased tau phosphorylation in female but not male mice. ConclusionsEpigenetic editing uncovered a female-specific role of Cdk5 activation in attenuating fear memory retrieval. This finding may be attributed to CDK5 mediated hyperphosphorylation of tau only in the female hippocampus. Sex-specific epigenetic regulation of Cdk5 may reflect differences in the effect of CDK5 on downstream target proteins that regulate memory.

Resveratrol epigenetically regulates the expression of zincfinger protein36 in non‑small cell lung cancer cell lines.

Zincfinger protein36 (ZFP36) is an AU‑rich element protein that binds to 3'‑untranslated regions and promotes the decay of target mRNAs. Downregulation of ZFP36 expression in turn results in stabilization of target mRNAs. A recent study indicated that downregulation of ZFP36 expression in human liver cancer is caused by epigenetic mechanisms. The purpose of the present study was to investigate the potential of resveratrol (Res) to induce ZFP36 expression. Promoter methylation was analyzed using methylation‑sensitive restriction analysis. It was determined that Res treatment increased ZFP36 expression and decreased the mRNA levels of ZFP36 target genes in A549 lung cancer cells. Additionally, Res suppressed the expression of DNA (cytosine‑5)‑methyltransferase1 and induced demethylation of the ZFP36 promoter. Collectively, the present results demonstrated that Res has anticancer activity through its epigenetic regulation of ZFP36 in non‑small cell lung cancer.

ZNF139 increases multidrug resistance in gastric cancer cells by inhibiting miR-185.

It has been reported that the expression of zinc finger protein 139 (ZNF139) and microRNA-185 (miR-185) were associated with proliferation, drug resistance of gastric cancer (GC) cells. However, the detailed mechanisms have not been fully investigated. The expression of ZNF139 in both GC tissues and cell lines was tested, then SGC7901/ADR or SGC7901 cells were transfected with ZNF139-siRNA, miR-185 analog, or pcDNA-ZNF139. Cell activity was determined by MTT assay. Real-time PCR and Western blot were utilized to detect ZNF139, miR-185, and multidrug resistance (MDR) related genes including MDR1/P-gp, GST-π, MRP-1, Bcl-2, TS and Bax. ChIP and dual luciferase activity assay were used to investigate regulation between ZNF139 and miR-185. Increased ZNF139 and decreased miR-185 expression were detected in GC tissues and cell lines. Transfection with ZNF139-siRNA into SGC7901/ADR cells markedly increased expression of miR-185, and treating with chemotherapeutic drugs ADR, 5-FU, L-OHP, the survival rate of SGC7901/ADR cells obviously decreased after ZNF139-siRNA transfection. On the other hand, transfection with pcDNA-ZNF139 in GC cell line SGC7901 resulted in an increased expression level of ZNF139 and a decline in the expression level of miR-185, meanwhile drug resistance of GC cells was clearly enhanced to ADR, 5-FU, L-OHP. Dual luciferase activity assay demonstrated that ZNF139 inhibited transcriptional activities of miR-185’s promoter in cells transfected with the reporter plasmid encompassing the upstream promoter region of miR-185 along with pcDNA-ZNF139. Our data reveal that ZNF139 might promote MDR gene MDR1/P-gp, MRP-1 and Bcl-2 by prohibiting miR-185.

Loss of Circulating Mucosal-Associated Invariant T Cells in Common Variable Immunodeficiency Is Associated with Immune Activation and Loss of Eomes and PLZF

Abstract Common variable immunodeficiency (CVID) is characterized by low levels of Igs leading to increased risk of infections. Mucosal-associated invariant T (MAIT) cells are a recently identified population of innate T cells with potent antibacterial activity. We hypothesized that CVID is associated with alterations in MAIT cells. Cryopreserved PBMC from CVID patients and healthy controls were used to study the frequency, phenotype, and response to Escherichia coli stimulation of MAIT cells by flow cytometry. MAIT cell frequency and absolute counts were depressed in CVID. Residual MAIT presented elevated coexpression of CD38 and HLA-DR, and reduced expression of CCR6, whereas levels of CD127 (IL-7 receptor) were unchanged. CVID patients also had an accumulation of MAIT cells lacking the critical transcription factors eomesodermin and promyelocytic leukemia zinc finger protein. MAIT cell frequency was inversely associated with levels of soluble CD14, with coexpression of CD38 and HLA-DR, and accumulation of MAIT cells lacking eomesodermin or promyelocytic leukemia zinc finger protein expression. None of these changes were normalized by IgG replacement therapy. Finally, MAIT cells from CVID patients displayed poor IFN-γ responses to E. coli stimulation, in part due to defective Ag presentation, and these responses were increased by pretreatment with IL-7. Defective MAIT cell response may contribute to the increased incidence of microbial infections seen in CVID patients on IgG replacement therapy.

Significance and expression of zinc finger protein 139 and matrix metalloproteinase 7 in endometrial adenocarcinoma

Objective To observe the expression of ZNF139 and MMP-7 in endometrial adenocarcinoma tissues and discuss the correlation with clinical pathological significances. Methods In 71 patients diagnosed as endometrial adenocarcinoma, 71 cancer tissues(adenocarcinoma group) and 71 adjacent normal tiuues(adjacent cancer group) were collected in operations.Immunohistochemical methods was used to detect the expression of ZNF139 and MMP-7 in the two groups, the relationship between the two genes and communication with clinicopathological indexes were discussed.Western blot and qRT-PCR were used to detected the expression of ZNF139 and MMP-7 at protein and mRNA level. Results The positive rates of ZNF139 was 66.20% in adenocarcinoma group, higher than in adjacent cancer group(21.42%, P=0.016); the positive rates of MMP-7 was 74.65% in adenocarcinoma group, higher than7 than in adjacent cancer group(26.76%, P=0.012). The expression of ZNF139 and MMP-7 was correlated with lymph node metastasis, muscle invasion and FIGO stage(P=0.028, 0.031; P=0.004, 0.016; P=0.008, 0.011). Spearman rank correlation analysis showed there was positive correlation between ZNF139 and MMP-7 in adenocarcinoma tissues(r=0.716, P=0.039). Western blot showed that ZNF139 and MMP-7 expression in adenocarcinoma group were higher than in adjacent cancer group(t=6.92, 7.34; P<0.01); qRT-PCR showed that ZNF139 mRNA and MMP-7 mRNA expression in adenocarcinoma group were higher than in adjacent cancer group(t=4.27, 4.06; P<0.05). Conclusion ZNF139 and MMP-7 are correlated with malignant pathological characteristics in endometrial adenocarcinoma, they may modulate the malignant behavior together. Key words: Endometrial adenocarcinoma; Zinc finger protein 139; Matrix metalloproteinase 7

Elevated Expression of Zinc Finger Protein 703 Promotes Cell Proliferation and Metastasis through PI3K/AKT/GSK-3β Signalling in Oral Squamous Cell Carcinoma

Zinc finger protein 703 (ZNF703), initially identified as a novel oncogene in human breast cancer, is a member of the NET/NlZ family of zinc finger transcription factors. It is recognized that the overexpression of ZNF703 is associated with various types of human cancers, but the role and molecular mechanism of ZNF703 in oral squamous cell carcinoma (OSCC) are unknown. ZNF703 expression levels were examined in OSCC tissues and non-cancerous tissues by qRT-PCR and immunohistochemistry (IHC). The molecular mechanisms of ZNF703 and its effects on cell growth and metastasis were explored in vitro and in vivo using the CCK8 assay, colony formation assay, cell cycle analysis, migration and invasion assays, wound-healing assay, western blotting and xenograft experiments in nude mice. In this study, ZNF703 was found to be upregulated in OSCC tissues compared to that in normal tissues at both mRNA and protein levels, and its expression level was closely correlated with the overall survival of patients with OSCC. Silencing of the ZNF703 gene in OSCC cells significantly inhibited cell growth and metastasis in vitro and in vivo. Conversely, the overexpression of ZNF703 in OSCC cells promoted cancer growth and metastasis in vitro. Mechanistically, ZNF703 activated the PI3K/AKT/GSK-3β signalling pathway and its downstream effectors, thus regulating the cell cycle and epithelial-mesenchymal transition (EMT). Furthermore, the promotive effects of ZNF703 on cellular proliferation and metastasis could be rescued by LY294002 (a PI3K-specific inhibitor) and MK2206 (an Akt-specific inhibitor). The results show that ZNF703 promotes cell growth and metastasis through PI3K/Akt/GSK-3β signalling in OSCC and that it may be a promising target in the treatment of patients with OSCC.

Maternal obesity epigenetically alters visceral fat progenitor cell properties in male offspring mice.

Maternal obesity reduces adipogenic progenitor density in offspring adipose tissue. The ability of adipose tissue expansion in the offspring of obese mothers is limited and is associated with metabolic dysfunction of adipose tissue when challenged with a high-fat diet. Maternal obesity induces DNA demethylation in the promoter of zinc finger protein 423, which renders progenitor cells with a high adipogenic capacity. Maternal obesity demonstrates long-term effects on the adipogenic capacity of progenitor cells in offspring adipose tissue, demonstrating a developmental programming effect. Maternal obesity (MO) programs offspring obesity and metabolic disorders, although the underlying mechanisms remain poorly defined. Progenitor cells are the source of new adipocytes. The present study aimed to test whether MO epigenetically predisposes adipocyte progenitors in the fat of offspring to adipogenic differentiation and subsequent depletion, which leads to a failure of adipose tissue plasticity under positive energy balance, contributing to adipose tissue metabolic dysfunction. C57BL/6 female mice were fed either a control diet (10% energy from fat) or a high-fat diet (45% energy from fat) for 8weeks before mating. Male offspring of control (Con) and obese (OB) dams were weaned onto a regular (Reg) or obesogenic (Obe) diet until 3months of age. At weaning, male OB offspring had a higher expression of Zinc finger protein 423 (zfp423), a key transcription factor in adipogenesis, as well as lower DNA methylation of its promoter in progenitors of epididymal fat compared to Con offspring, which was correlated with enhanced adipogenic differentiation. At 3months of age, progenitor density was 30.9±9.7% lower in OB/Obe compared to Con/Obe mice, accompanied by a limited expansion of the adipocyte number when challenged with a high-energy diet. This difference was associated with lower DNA methylation in the zfp423 promoter in the epididymal fat of OB/Obe offspring, which was correlated with greater macrophage chemotactic protein-1 and hypoxia-inducible factor 1α expression. In summary, MO epigenetically limits the expansion capacity of offspring adipose tissue, providing an explanation for the adipose metabolic dysfunction of male offspring in the setting of MO.

Regulation of the expression of zinc finger protein genes by microRNAs enriched within acute lymphoblastic leukemia-derived microvesicles

Microvesicles (MVs) are submicrometric membrane fragments that can "engulf" cytoplasmic contents such as microRNAs (miRNAs) from their cellular origin. The study of miRNAs carried within MVs might provide insights into the roles that miRNAs play in the underlying pathophysiologic processes of acute lymphoblastic leu-kemia (ALL). We identified numerous dysregulated MV miRNAs in patients with B- and T-cell ALL by using Agilent microarray analysis. Selected miRNAs obtained by microarray profiling were validated us-ing quantitative reverse transcription-polymerase chain reaction. Us-ing bioinformatic tools, we found that 118 and 116 miRNAs from B- and T-ALL MVs, respectively, regulated the expression of zinc finger protein (ZFP) genes. For example, zinc finger protein 238 (ZNF238), known as a tumor suppressor, was regulated by miR-20b over-expres-sion. Conversely, ZNF267, a cancer-promoting factor, was mediated by downregulated miR-23a and miR-23b. Considering that miRNAs are generally believed to repress gene expression, antineoplastic ZNF238 was likely inhibited while the level of oncogenic ZNF267 was likely increased by miRNA dysregulation, leading to modifica-tion of the ALL microenvironment. In addition, gene ontology and sig-naling pathway analysis demonstrated that a subset of the ZFP genes targeted by altered MV miRNAs are involved in cellular biological processes including proliferation, differentiation, apoptosis, and cell cycle regulation. These findings indicated that cancer-associated MV miRNAs and their target ZFP genes might be novel pathogenic factors in ALL. However, the specific roles exerted by MV miRNAs and their target ZFP genes on the pathological mechanisms of ALL remain to be further understood.

Frequent expression of zinc-finger protein ZNF165 in human urinary bladder transitional cell carcinoma

The aim of the study is to evaluate mRNA/protein expression of zinc finger protein 165 (ZNF165) in transitional cell carcinomas (TCCs) of urinary bladder and correlate its expression with the clinicopathological characteristics of patients. In this study, the methods of quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemistry (IHC) were utilized to evaluate mRNA/protein expression of ZNF165 in TCC. Independent Student's t test, ANOVA and Chi-square (χ2) were used to analyze the data statistically. We observed overexpression of ZNF165 mRNA in testis and majority (59.2%) of TCC patients. ZNF165 mRNA expression was also detected in adjacent noncancerous tissues (ANCTs) and some other normal tissues. Relative mean fold expression of ZNF165 mRNA was found to be significantly (p<0.01) higher in muscle-invasive bladder cancer (MIBC) as compared to non-muscle-invasive bladder cancer (NMIBC) patients. (12.11±9.57 vs. 5.72±2.61, p=0.009). ZNF165 protein expression was demonstrated on archival formalin-fixed, paraffin-embedded (FFPE) bladder tissues using IHC and nuclear staining pattern was detected. No significant difference was observed in protein expression of ZNF165 between the two groups (NMIBC and MIBC patients) (61.1% vs. 55.2%, p=0.629). No significant protein expression of ZNF165 was observed among ANCTs and benign prostatic hyperplasia (BPH) used as control. Our study results suggest that ZNF165 mRNA/protein expression was observed in TCC of human urinary bladder and might be used as a novel diagnostic biomarker and as well a vaccine target in development of urinary bladder cancer specific immunotherapy.

Maternal overnutrition enhances mRNA expression of adipogenic markers and collagen deposition in skeletal muscle of beef cattle fetuses.

Twenty-four pregnant Nellore cows were randomly assigned into 2 feeding level groups (control [CTL]; fed 1.0 times the maintenance requirement; n = 12; and overnourished [ON]; fed at 1.5 times the maintenance requirement; n = 12) to evaluate effects of maternal overnutrition on fetal skeletal muscle development. Cows were slaughtered at 135, 190, and 240 d of gestation and samples of fetal LM were collected for analysis of mRNA expression analysis and for histological evaluation of collagen content and number of muscle cells. There was no interaction between gestational period and maternal nutrition for the variables evaluated (P > 0.05). The mRNA expression of Cadherin-associated protein, β 1 (β-catenin) tended to be greater in fetuses from ON cows (P = 0.08), while myogenic differentiation 1 (MyoD; P = 0.56), myogenin (MyoG; P = 0.70), and the number of muscle cells (P = 0.90) were not affected by maternal overnutrition. Gestational period did not affect the mRNA expression of β-catenin (P = 0.60) and MyoG (P = 0.21). The mRNA expression of MyoD tended to increase with days of gestation (P = 0.06). The mRNA expression of zinc finger protein 423 (Zfp423; P < 0.0001), C/EBPα (P = 0.01), and PPARγ (P < 0.0001) were enhanced in ON fetuses. No effects of days of gestation were observed for mRNA expression of Zfp423 (P = 0.75) and C/EBPα (P = 0.48). The mRNA expression of PPARγ in fetuses at 190 d of gestation tended to be greater than those at 135 and 240 d of gestation (P = 0.06). The mRNA expression of transforming growth factor β (TGF-β; P < 0.0001), collagen type III, α I (COL3A1; P < 0.0001), and collagen content (P = 0.01) were increased in ON fetuses. Gestational period did not affect the mRNA expression of collagen type I, α I (COL1A1; P = 0.65). The mRNA expression of COL3A1 (P = 0.09) in fetuses at 190 d of gestation tended to be greater than fetuses at 135 and 240 d of gestation. The mRNA expression of TGF-β in fetuses at 190 d of gestation was greater than in fetuses at 135 d of gestation (P = 0.03), and the values observed in fetuses at 240 d of gestation did not differ from the other gestational time points. The least value of collagen content (P = 0.01) was observed in fetuses at 135 d of gestation, and no differences were observed among the other gestational time points. These data shows that maternal overnutrition enhances fibrogenesis and likely adipogenesis without compromising myogenesis in fetal skeletal muscle of cattle.

Identification of expression profiles associated to oral squamous cell carcinoma development by cDNA microarrays (1047.13)

Oral squamous cell carcinoma (OSCC) development still a poorly understood phenomenon, in terms of gene expression profiles which might be involved in alterations leading to high invasiveness and aggressive behavior. Also, it stills unclear how gene expression changes during OSCC development, in terms of tumor size. Using 36 samples of OSCC cases, we performed cDNA and lincRNA microarrays, comprising whole human genome and more than 7000 lincRNAs, in order to identify gene profiles with similar behavior throughout T1 to T4 tumor staging. Also, 10 differentially expressed genes were tested individually using qRT‐PCR. We found 7 different gene expression profiles, either with a rise in expression levels, or decrease in expression levels, between T1, T2, T3 and T4 TNM staging. The seven different gene expression profiles showed 58 differentially expressed genes, including an increased expression of immune response‐related family genes, and decreased expression of zinc finger proteins. In RT‐PCR experiments, we found significantly correlations between CD274 (PD‐L1) and tumor size (p=0.043), lymphatic invasion (p=0.047) and metastatic lymph node (p=0.001); BLNK and perineural invasion (p=0.021); ABL2 and tumor size (p=0.039); HOXB9 and tumor size (p=0.026); and between ZNF813 and tumor size (p=0.039). Our study shows the existence of several genes with similar expression patterns during OSCC development, and also points to novel genes, which has not yet been implicated in oral carcinogenesis, providing new potential targets in oral cancer research.Grant Funding Source: Supported by FAPESP grants 2010/08637‐0 and 2010/08400‐0

Zinc finger protein 139 expression in gastric cancer and its clinical significance

To investigate the expression of zinc finger protein 139 (ZNF139) in gastric cancer (GC), and to analyze its clinical significance. A total of 108 patients who were diagnosed with GC and underwent surgery between January 2005 and March 2007 were enrolled in this study. Gastric tumor specimens and paired tumor-adjacent tissues were collected and paraffin-embedded, and the clinicopathologic characteristics and prognosis were recorded. The expression of ZNF139, Bcl-2, Bax, and caspase-3 were determined by immunohistochemistry, and apoptosis was assessed by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling. SPSS 13.0 software was used for data processing and analyses, and significance was determined at P < 0.05. The expression of ZNF139 was stronger in tumors than in tumor-adjacent tissues (66.67% vs 44.44%; P < 0.01). Overexpression of ZNF139 correlated with tumor differentiation, invasion depth, clinical stage, lymphatic metastasis, and blood vessel invasion (all Ps < 0.05). Patients with overexpression of ZNF139 had a poorer prognosis (P < 0.01), and overexpression of ZNF139 was an independent factor for the prognosis of GC patients by a Cox survival analysis (P = 0.02). A negative relationship between ZNF139 and the apoptosis index was observed (r = -0.686; P < 0.01). The expression of Bcl-2 in GC was stronger than in tumor-adjacent tissues (66.67% vs 41.67%), whereas the expression levels of Bax and caspase-3 were lower in primary tumors (54.63% and 47.22%, respectively) than in tumor-adjacent tissues (73.15% and 73.15%, respectively) (all Ps < 0.05). The expression of ZNF139 negatively correlated with caspase-3 (r = -0.370; P < 0.01). The expressions of Bcl-2 and Bax were also negatively correlated (r = -0.231; P = 0.02). The expressions of caspase-3 and Bax protein were positively correlated (r = 0.217; P = 0.024). ZNF139 is related to clinicopathologic characteristics and prognosis of GC. Furthermore, it is overexpressed and involved in apoptosis in GC tissues by regulating caspase-3.

Differential expression of zinc finger proteins 36 (ZFP36s) family and leukemia inhibitory factor (LIF) in human endometrial epithelium and stroma

The ZFP36 family (ZFP36s) of RNA-binding proteins (ZFP36 (TTP), ZFP36L1 (L1), ZFP36L2 (L2)) regulates gene expression. Evidence in embryonic fibroblasts suggests that ZFP36s modulates expression of LIF, a putative endometrial regulator critical for embryo implantation. We previously reported a complex relationship between the expression of ZFP36s and LIF in whole human endometrium. The purpose of this study was to examine expression in separated human endometrial epithelium and stroma.

Gene Profiles during Vestibular Compensation in Rats after Unilateral Labyrinthectomy

The aims of this study were to determine changes in gene expression in the chronic state of vestibular compensation by microarray analysis and to validate the asymmetrical levels of gene expression in the ipsilateral and contralateral vestibular nucleus complexes (VNCs). Microarray analysis was used to examine the expression of genes up-regulated or down-regulated in the ipsilateral VNC at 1 and 7 days after unilateral labyrinthectomy. Up-regulated or down-regulated gene expression in the ipsilateral and contralateral VNCs was then validated by reverse transcriptase polymerase chain reaction at 1, 7, 14, and 28 days after labyrinthectomy. The genes down-regulated at 1 day after labyrinthectomy and up-regulated at 7 days after labyrinthectomy as determined by microarray analysis and reverse transcriptase polymerase chain reaction were zinc finger protein 307, zinc metallopeptidase, P34, calcitonin receptor, insulin-like growth factor binding protein 5, GATA binding protein 3, and CD151. Expression of zinc finger protein 307, zinc metallopeptidase, P34, and calcitonin receptor was up-regulated even after 7 days in the contralateral VNC of rats that had labyrinthectomy. This study demonstrated changes in gene expression in rats during the chronic phase of vestibular compensation after unilateral labyrinthectomy and provided profiles of these changes in gene expression.

Identification of Regulatory Foxp3+ Invariant NKT Cells Induced by TGF-β

Invariant NKT (iNKT) cells were shown to prevent the onset of experimental autoimmune encephalomyelitis in mice following administration of their specific TCR agonist alpha-galactosylceramide. We found that this protection was associated with the emergence of a Foxp3(+) iNKT cell population in cervical lymph nodes. We demonstrate that the differentiation of these cells is critically dependent on TGF-beta in both mice and humans. Moreover, in vivo generation of Foxp3(+) iNKT cells was observed in the TGF-beta-rich environment of the murine gut. Foxp3(+) iNKT cells displayed a phenotype similar to that of Foxp3(+) regulatory T cells, and they suppress through a contact-dependent, glucocorticoid-induced TNFR-mediated mechanism. Nevertheless, Foxp3(+) iNKT cells retain distinctive NKT cell characteristics, such as promyelocytic leukemia zinc finger protein expression and preferential homing to the liver following adoptive transfer, where they stably maintained Foxp3 expression. Our data thus unveil an unexpected capacity of iNKT cells to acquire regulatory functions that may contribute to the establishment of immunological tolerance.