Fox transcription factors play a critical role in the regulation of a variety of biological processes. While FoxM1 behaves like the oncogenic transcription factor, FoxO3a is known as a tumor suppressor by inhibiting FoxM1. This study aimed to investigate the clinicopathological significance of FoxM1 and FoxO3a expression in breast cancer. Expression of FoxM1 and FoxO3a were analyzed by immunohistochemical staining on tissue microarray sections from 236 breast cancer patients, and correlated with various clinicopathological characteristics. Overexpression of FoxM1 correlated with adverse clinicopathological features, such as larger tumor size, lymph node metastasis, advanced tumor stage, and lymphovascular invasion. The Kaplan-Meier survival curves revealed no prognostic significance of FoxM1 expression. However, in subgroup analyses with patients of estrogen receptor (ER) positive breast cancers, FoxM1 overexpression associated with poor disease free and overall survival. No association was found between FoxO3a and FoxM1 expression. Regarding clinicopathological variables, the only association between histologic grade and FoxO3a was observed. In conclusion, FoxM1 overexpression was significantly associated with aggressive phenotypes and poor prognosis of ER-positive breast cancer. These findings suggest the possible role of FoxM1 as a prognostic biomarker and putative target of anti-cancer therapy.
Breast Neoplasms/chemistry/mortality/*pathology ; Female ; Forkhead Transcription Factors/*analysis ; Humans ; Phenotype ; Prognosis ; Receptor, ErbB-2/analysis ; Receptors, Estrogen/*analysis

Numerous studies have reported that genes with similar expression patterns are co-regulated. From gene expression data, we have assumed that genes having similar expression pattern would share similar transcription factor binding sites (TFBSs). These function as the binding regions for transcription factors (TFs) and thereby regulate gene expression. In this context, various analysis tools have been developed. However, they have shortcomings in the combined analysis of expression patterns and significant TFBSs and in the functional analysis of target genes of significantly overrepresented putative regulators. In this study, we present a web-based A Functional Clustering Analysis Tool for Predicted Transcription Regulatory Elements and Gene Ontology Terms (FCAnalyzer). This system integrates microarray clustering data with similar expression patterns, and TFBS data in each cluster. FCAnalyzer is designed to perform two independent clustering procedures. The first process clusters gene expression profiles using the K-means clustering method, and the second process clusters predicted TFBSs in the upstream region of previously clustered genes using the hierarchical biclustering method for simultaneous grouping of genes and samples. This system offers retrieved information for predicted TFBSs in each cluster using Match(TM) in the TRANSFAC database. We used gene ontology term analysis for functional annotation of genes in the same cluster. We also provide the user with a combinatorial TFBS analysis of TFBS pairs. The enrichment of TFBS analysis and GO term analysis is statistically by the calculation of P values based on Fisher's exact test, hypergeometric distribution and Bonferroni correction. FCAnalyzer is a web-based, user-friendly functional clustering analysis system that facilitates the transcriptional regulatory analysis of co-expressed genes. This system presents the analyses of clustered genes, significant TFBSs, significantly enriched TFBS combinations, their target genes and TFBS-TF pairs.
Binding Sites ; Cluster Analysis* ; Gene Expression ; Gene Ontology* ; Transcription Factors ; Transcriptome

OBJECTIVETo probe into thinking and prospect of application of gene chip technique to research of the channel-viscera correlativity.

METHODSAdopt literature analytic method, review and analyze current state of application of gene chip in biology and medicine, and achievements in studies of channel-viscera correlativity.

RESULTSIn biological and medical fields, gene chip can be applied to high flux expression parallel analysis, large-scale gene discovery and gene analysis, gene polymorphous analysis, genome study, etc, especially, it has important application values in gene expression map; channel-viscera correlativity need stereo-crossing studies by multiple subjects, multiple systems, multiple directions and multiple levels, being more systematic and sequential, and gene chip technique is applicable to this requirement.

CONCLUSIONWide application of gene chip technique will comprehensively promote deep study of channel-viscera correlativity.

Gene Expression ; Humans ; Oligonucleotide Array Sequence Analysis ; Transcription Factors ; metabolism ; Viscera

BACKGROUND: SIRT7 is one of the histone deacetylases and is NAD-dependent. It forms a complex with ETS-like transcription factor 4 (ELK4), which deacetylates H3K18ac and works as a transcriptional suppressor. Overexpression of SIRT7 and deacetylation of H3K18ac have been shown to be associated with aggressive clinical behavior in some cancers, including hepatocellular carcinoma (HCC). The present study investigated the immunohistochemical expression of SIRT7, H3K18ac, and ELK4 in hepatocellular carcinoma. METHODS: A total of 278 HCC patients were enrolled in this study. Tissue microarray blocks were made from existing paraffin-embedded blocks. Immunohistochemical expressions of SIRT7, H3K18ac and ELK4 were scored and analyzed. RESULTS: High SIRT7 (p = .034), high H3K18ac (p = .001), and low ELK4 (p = .021) groups were associated with poor outcomes. Age < 65 years (p = .028), tumor size ≥ 5 cm (p = .001), presence of vascular emboli (p = .003), involvement of surgical margin (p = .001), and high American Joint Committee on Cancer stage (III&V) (p < .001) were correlated with worse prognoses. In multivariate analysis, H3K18ac (p = .001) and ELK4 (p = .015) were the significant independent prognostic factors. CONCLUSIONS: High SIRT7 expression with poor overall survival implies that deacetylation of H3K18ac contributes to progression of HCC. High H3K18ac expression with poor prognosis is predicted due to a compensation mechanism. In addition, high ELK4 expression with good prognosis suggests another role of ELK4 as a tumor suppressor beyond SIRT7's helper. In conclusion, we could assume that the H3K18ac deacetylation pathway is influenced by many other factors.
Carcinoma, Hepatocellular* ; Compensation and Redress ; Histone Deacetylases ; Humans ; Immunohistochemistry ; Joints ; Multivariate Analysis ; Prognosis ; Transcription Factors

Postnatal heart maturation is the basis of normal cardiac function and provides critical insights into heart repair and regenerative medicine. While static snapshots of the maturing heart have provided much insight into its molecular signatures, few key events during postnatal cardiomyocyte maturation have been uncovered. Here, we report that cardiomyocytes (CMs) experience epigenetic and transcriptional decline of cardiac gene expression immediately after birth, leading to a transition state of CMs at postnatal day 7 (P7) that was essential for CM subtype specification during heart maturation. Large-scale single-cell analysis and genetic lineage tracing confirm the presence of transition state CMs at P7 bridging immature state and mature states. Silencing of key transcription factor JUN in P1-hearts significantly repressed CM transition, resulting in perturbed CM subtype proportions and reduced cardiac function in mature hearts. In addition, transplantation of P7-CMs into infarcted hearts exhibited cardiac repair potential superior to P1-CMs. Collectively, our data uncover CM state transition as a key event in postnatal heart maturation, which not only provides insights into molecular foundations of heart maturation, but also opens an avenue for manipulation of cardiomyocyte fate in disease and regenerative medicine.
Gene Expression Regulation ; Heart ; Myocytes, Cardiac/metabolism* ; Single-Cell Analysis ; Transcription Factors/metabolism*

OBJECTIVETo explore the molecular etiology of two pedigrees affected with type II Waardenburg syndrome (WS2) and to provide genetic diagnosis and counseling.

METHODSBlood samples were collected from the proband and his family members. Following extraction of genomic DNA, the coding sequences of PAX3, MITF, SOX10 and SNAI2 genes were amplified with PCR and subjected to DNA sequencing to detect potential mutations.

RESULTSA heterozygous deletional mutation c.649_651delAGA in exon 7 of the MITF gene has been identified in all patients from the first family, while no mutation was found in the other WS2 related genes including PAX3, MITF, SOX10 and SNAI2.

CONCLUSIONThe heterozygous deletion mutation c.649_651delAGA in exon 7 of the MITF gene probably underlies the disease in the first family. It is expected that other genes may also underlie WS2.

Base Sequence ; DNA Mutational Analysis ; Exons ; genetics ; Family Health ; Female ; Genetic Predisposition to Disease ; genetics ; Heterozygote ; Humans ; Male ; Microphthalmia-Associated Transcription Factor ; genetics ; Molecular Sequence Data ; Mutation ; PAX3 Transcription Factor ; Paired Box Transcription Factors ; genetics ; Pedigree ; Polymerase Chain Reaction ; SOXE Transcription Factors ; genetics ; Sequence Deletion ; Snail Family Transcription Factors ; Transcription Factors ; genetics ; Waardenburg Syndrome ; classification ; diagnosis ; genetics

BACKGROUNDHuman embryonic stem (HES) cell derived from human blastocyst can be propagated indefinitely in the primitive undifferentiated state while remaining pluripotent. It has exciting potential in human developmental biology, drug discovery, and transplantation medicine. But there are insufficient HES cell lines for further study.

METHODSThree oocyte donors were studied, and 3 in vitro fertilization (IVF) cycles were carried out to get blastocysts for the establishment of HES cell line. Isolated from blastocysts immunosurgically, inner cell mass (ICM) was cultured and propagated on mouse embryonic fibroblasts (MEFs). Once established, morphology, cell surface markers, karyotype and differentiating ability of the cell line were thoroughly analyzed.

RESULTSFour ICMs from 7 blastocysts were cultured on MEFs. After culture, one cell line (cHES-1) was established and met the criteria for defining human pluripotent stem cells including a series of markers used to identify pluripotent stem cells, morphological similarity to primate embryonic stem cells and HES reported else where. Normal and stable karyotype maintained over 60 passages, and demonstrated ability to differentiate into a wide variety of cell types.

CONCLUSIONSHES cell lines can be established from gamete donors at a relatively highly efficient rate. The establishment will exert a widespread impact on biomedical research.

Blastocyst ; cytology ; Cell Differentiation ; Cell Line ; DNA-Binding Proteins ; analysis ; Female ; Fertilization in Vitro ; Humans ; Karyotyping ; Male ; Octamer Transcription Factor-3 ; Stem Cells ; cytology ; Tissue Donors ; Transcription Factors ; analysis

BACKGROUNDT cell factor-4 (TCF-4) plays an important role in development and carcinogenesis. Recently, the role of TCF-4 has been described in colon cancer and other cancers. However, whether TCF-4 plays a similar role in lung cancer is unknown. To answer this question, we studied the expression of TCF-4 protein and mRNA in non-small-cell lung cancer (NSCLC) and the relation of TCF-4 expression pattern to histological type and cell differentiation.

METHODSTissue samples from sixty cases of pathologically diagnosed NSCLC and eight normal tissue samples were obtained between September 2001 and March 2003. Immunohistochemistry was used to investigate the distribution of TCF-4 protein. The staining patterns of the tumors were divided into 4 categories: nuclear staining alone or nuclear staining greater than cytoplasmic staining; cytoplasmic staining or cytoplasmic staining greater than nuclear staining; equal nuclear and cytoplasmic staining; no nuclear staining or cytoplasmic staining. The integrated optical density (OD) values of all sections were analyzed by UIC MetaMorph image analysis software. The expression of TCF-4 mRNA was detected by one-step reverse transcription-polymerase chain reaction (RT-PCR). The integrated density values of the PCR products were analyzed semi-quantitatively.

RESULTSImmunohistochemistry showed that there was no expression of TCF-4 in normal tissue. However, TCF-4 was expressed in 86.7% (52/60) of NSCLC samples, mainly in the nuclei of tumor cells. Furthermore, there was a significant difference in TCF-4 localization patterns between squamous cell carcinomas and adenocarcinomas (P < 0.05). The integrated OD values of TCF-4 expression was significantly higher in tumors with moderate-poor cell differentiation than in well differentiated tumors (51.63 +/- 6.67 vs 46.13 +/- 12.31, P < 0.01). There was no TCF-4 mRNA expression in normal tissue. However, 63.9% (23/36) of carcinoma samples expressed TCF-4 mRNA. TCF-4 mRNA expression was significantly higher in tumors with moderate-poor cell differentiation than in well differentiated tumors (P < 0.05). There were no significant differences in mRNA expression in comparison with histological type.

CONCLUSIONSThe sub-cellular distribution of TCF-4 may correlate with NSCLC histological type. High expression of TCF-4 mRNA and protein may be associated with the degree of cell differentiation in NSCLC.

Carcinoma, Non-Small-Cell Lung ; chemistry ; Cytoskeletal Proteins ; metabolism ; Female ; Humans ; Immunohistochemistry ; Lung Neoplasms ; chemistry ; Male ; Middle Aged ; RNA, Messenger ; analysis ; TCF Transcription Factors ; Trans-Activators ; metabolism ; Transcription Factor 7-Like 2 Protein ; Transcription Factors ; analysis ; genetics ; beta Catenin

OBJECTIVETo study how the combined effects of various differentiation inducers and heat stress on the expression of JWA protein in K562 cell, the relationship between JWA and Hsp70 expression, and the signal regulation mechanism possibly involved.

METHODSThe experimental model was established in K562 cells. Various directional differentiation inducers (TPA, Ara-C, hemin, adriamycin, ATRA and As(2)O(3)) were used alone or combined with heat shock treatment (42 degrees C, 2 h). Western blot was used for detecting expressions of JWA, Hsp70, heat stress factor 1 (HSF1) and HSF2.

RESULTS(1) The expressions of both JWA protein and Hsp70 were significantly up-regulated after K562 cells treated by TPA (100, 200 ng/ml) or adriamycin (4 x 10(-8) mol/L) 48 h, and followed by heat shock (42 degrees C, 2 h). However, the opposite effects were observed when the cells treated by hemin (3 x 10(-5) mol/L, 48 h), Ara-C (80 ng/ml, 48 h) and As(2)O(3) (1 x 10(-6) mol/L, 48 h) followed by 2 h heat shock. No obvious changes were found when the cells treated by ATRA (1 x 10(-6) mol/L, 48 h) alone or followed by heat shock. (2) Both the heat shock transcriptional factors HSF1 and HSF2 did not show any significant changes when K562 cells were treated with various differentiation inducers and followed by heat stress.

CONCLUSIONJWA not only takes part in the regulation of K562 cellular differentiation, but also of heat stress, it might be the co-target gene of several differentiation inducers and heat stress. The expression of Hsp70 seems not mediated by both HSF1 and HSF2 in K562 cells undergoing directional differentiation or heat stress treatment. JWA is likely to be a new signal molecule similar to Hsp70 signal pathways. The results show that JWA takes part in the mechanism of K562 cell response to heat stress.

Blotting, Western ; DNA-Binding Proteins ; analysis ; Flow Cytometry ; HSP70 Heat-Shock Proteins ; analysis ; Heat Shock Transcription Factors ; Heat-Shock Proteins ; analysis ; Hot Temperature ; Humans ; Intracellular Signaling Peptides and Proteins ; K562 Cells ; Transcription Factors ; analysis

BACKGROUNDNurr1 is a member of the nuclear receptor superfamily of transcription factors. The objective of the present study was to identify novel splicing variants of the gene in neuronal and non-neuronal tissues and determine their functions.

METHODSReverse transcription-polymerase chain reaction (RT-PCR) analysis was used to screen for Nurr1 splice variants in the adult human central nervous system (CNS) and in other tissues such as lymphocytes, and liver, muscle, and kidney cells. Functional assays of the variants were performed by measuring Nurr1 response element (NuRE) transcriptional activity in vitro.

RESULTSIn this study, the authors identified a novel splicing variant of Nurr1 within exon 5, found in multiple adult human tissues, including lymphocytes, and liver, muscle, and kidney cells, but not in the brain or spinal cord. Sequencing analysis showed the variant has a 75 bp deletion between nucleotides 1402 and 1476. A functional assay of the Nurr1-c splicing variant, performed by measuring NuRE transcriptional activity in vitro, detected a 39% lower level of luciferase (LUC) activity (P < 0.05).

CONCLUSIONA novel splicing variant of Nurr1 exists in human non-neuronal tissues and functional assays suggest that the variant may act as an alternate transcription regulator.

Adult ; Alternative Splicing ; DNA-Binding Proteins ; analysis ; genetics ; Humans ; Nuclear Receptor Subfamily 4, Group A, Member 2 ; Transcription Factors ; analysis ; genetics