To determine the feasibility of a nonradioactive electrophoresis mobility shift assay for detecting nuclear transcription factor, double-stranded oligonucleotides encoding the consensus target sequence of NF-kappa B were labelled with DIG by terminal transferase. After nuclear protein stimulated with phorbol 12-myristate 13-acetate (PMA) or PMA and pyrrolidine dithiocarbamate (PDTC) electrophoresed on 8% nondenaturing poliacrylamide gel together with oligeonucleotide probe, they were electro-blotted nylon membrane positively charged. Anti-DIG-AP antibody catalyzed chemiluminescent substrate CSPD to image on X-film. The results showed that nuclear proteins binded specifically to the NF-kappa B consensus sequence in the EMSA by chemiluminescent technique method and the activity of NF-kappa B in PMA group was more than that in PMA + PDTC group. It is suggested that detection of NF-kappa B by EMSA with chemiluminescent technique is feasible and simple, which can be performed in ordinary laboratories.
Chemiluminescent Measurements ; DNA-Binding Proteins/*analysis ; DNA-Binding Proteins/genetics ; Electrophoretic Mobility Shift Assay ; NF-kappa B/*analysis ; NF-kappa B/genetics ; NF-kappa B/metabolism ; Rats, Sprague-Dawley ; *Trans-Activation (Genetics) ; Trans-Activators/analysis ; Trans-Activators/genetics ; *Transcription, Genetic

No abstract available.
Acetyltransferases/metabolism ; Animal ; Carrier Proteins/metabolism ; DNA-Binding Proteins/metabolism ; Human ; Nuclear Proteins/metabolism ; Receptors, Cytoplasmic and Nuclear/metabolism* ; Repressor Proteins/metabolism*n ; Trans-Activators/metabolism* ; Transcription Factors/metabolism

<b>OBJECTIVEb>To study the clinical significance of P53, C-MYC and BCL-6 abnormality in the patients with diffuse large B cell lymphoma (DLBCL).

<b>METHODSb>From July 2011 to January 2013, 80 patients with DLBCL were admitted in our hospital and were chosen as study objects, their clinical data were collected. The abnormality of P53, C-MYC and BCL-6 was examined by using I-FISH for all the patients. The correlation of abnormality of P53, C-MYC and BCL-6 with clinical staging, curative efficacy and prognosis of the patients were analyzed.

<b>RESULTSb>Out of 80 patients 27 patients (33.75%) had P53 deletion, 24 patients (30.00%) had C-MYC rearrangement/amplification, and 46 patients (57.50%) had BCL-6 rearrangement. The P53 deletion, C-MYC rearrangement/amplification and BCL-6 rearrangement significantly correlated with staging, curative effect and prognosis of the patients (P < 0.05).

<b>CONCLUSIONb>The curative efficacy and prognosis of the DLBCL patients with abnormality of P53, C-MYC and BCL-6 have been confirmed to be unsatisfactory.

DNA-Binding Proteins ; genetics ; metabolism ; Humans ; In Situ Hybridization, Fluorescence ; Lymphoma, Large B-Cell, Diffuse ; diagnosis ; genetics ; metabolism ; Prognosis ; Proto-Oncogene Proteins c-bcl-6 ; Proto-Oncogene Proteins c-myc ; genetics ; metabolism ; Tumor Suppressor Protein p53 ; genetics ; metabolism

WT1 gene encodes a zinc finger transcription factor that regulates transcription of its downstream genes. Some of target genes for WT1 are involved in regulating both cell cycle and cellular proliferation and differentiation. However, WT1 itself is regulated by its upstream genes such as NF-kappaB and GATA-1. Thus there exists a pathway of transcriptional regulation mediated by WT1, which controls development of hematopoietic system. Leukemia results from disrupting the homeostasis among hematopoietic proliferation, differentiation and apoptosis, which is often the consequence of an inappropriate expression of transcription factors and subsequent disruption of the normal gene expression pattern. This article reviews the relationship between the WT1-mediated pathway of transcriptional regulation and leukemia.
Animals ; Carrier Proteins ; genetics ; Cyclin-Dependent Kinase Inhibitor p21 ; Cyclins ; genetics ; DNA-Binding Proteins ; metabolism ; Erythroid-Specific DNA-Binding Factors ; GATA1 Transcription Factor ; Gene Expression Regulation ; Humans ; Leukemia ; etiology ; genetics ; NF-kappa B ; metabolism ; Nuclear Proteins ; genetics ; Retinoblastoma-Binding Protein 7 ; Transcription Factors ; metabolism ; Transcription, Genetic ; WT1 Proteins ; physiology

<b>OBJECTIVEb>To investigate the effect of stat3 phosphorylation and p53 expression on human epidermal non-melanoma cutaneous tumours.

<b>METHODSb>Immunohistochemistry technique was employed to measure the expression of p-stat3 and p53 protein in skin tissue from 30 cases of skin squamous cell carcinoma (SCC), 20 cases of basal cell carcinoma (BCC), 20 cases of seborrhoeic keratosis (SK) and 20 normal subjects.

<b>RESULTb>(1) p-stat3 protein was abnormally increased in SCC and BCC as compared with normal skin and SK. Expression of p-stat3 in SCC was also significantly higher than that in BCC. (2) Expression of p-stat3 was higher in poorly-differentiated cancers than that in well-differentiated cancers in SCC. The positive rate of p-stat3 expression was correlated with the depth of tumor invasion, but not with tumor size. (3) There was no p53 protein expression on normal skin and SK, it was significantly upregulated in SCC and BCC. In SCC, the intensity of p53 expression was associated with tumor differentiation. There was no correlation between the positive rate of p53 expression and the depth of tumor invasion, whereas the positive rate of p53 expression was correlated with the sun-exposure area. (4) There existed positive correlation between the expression intensity of p-stat3 and p53 in SCC (r=0.641, P<0.05).

<b>CONCLUSIONb>(1) The overexpression of p-stat3 may play an important role in the development of epidermal tumors. (2) The abnormal activation of stat3 may be related to metastatic potentials in SCC. (3) Both p53 gene and stat3 may contribute to the pathogenesis of skin SCC.

Carcinoma, Basal Cell ; chemistry ; pathology ; Carcinoma, Squamous Cell ; chemistry ; pathology ; DNA-Binding Proteins ; metabolism ; Humans ; Immunohistochemistry ; Keratosis, Seborrheic ; metabolism ; Phosphorylation ; STAT3 Transcription Factor ; Skin ; chemistry ; Skin Neoplasms ; chemistry ; pathology ; Trans-Activators ; metabolism ; Tumor Suppressor Protein p53 ; analysis

To determine the feasibility of a nonradioactive electrophoresis mobility shift assay for detecting nuclear transcription factor, double-stranded oligonucleotides encoding the consensus target sequence of NF-kappa B were labelled with DIG by terminal transferase. After nuclear protein stimulated with phorbol 12-myristate 13-acetate (PMA) or PMA and pyrrolidine dithiocarbamate (PDTC) electrophoresed on 8% nondenaturing poliacrylamide gel together with oligeonucleotide probe, they were electro-blotted nylon membrane positively charged. Anti-DIG-AP antibody catalyzed chemiluminescent substrate CSPD to image on X-film. The results showed that nuclear proteins binded specifically to the NF-kappa B consensus sequence in the EMSA by chemiluminescent technique method and the activity of NF-kappa B in PMA group was more than that in PMA + PDTC group. It is suggested that detection of NF-kappa B by EMSA with chemiluminescent technique is feasible and simple, which can be performed in ordinary laboratories.
Animals ; DNA-Binding Proteins ; analysis ; genetics ; Electrophoretic Mobility Shift Assay ; Luminescent Measurements ; NF-kappa B ; analysis ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Trans-Activators ; analysis ; genetics ; Transcription, Genetic ; Transcriptional Activation

Cellular senescence is an irreversible cell cycle arrest triggered by the activation of oncogenes or mitogenic signaling as well as the enforced expression of tumor suppressors such as p53, p16(INK4A) and promyelocytic leukemia protein (PML) in normal cells. E2F-binding protein 1 (E2FBP1), a transcription regulator for E2F, induces PML reduction and suppresses the formation of PML-nuclear bodies, whereas the down-regulation of E2FBP1 provokes the PML-dependent premature senescence in human normal fibroblasts. Here we report that the depletion of E2FBP1 induces the accumulation of PML through the Ras-dependent activation of MAP kinase signaling. The cellular levels of p16(INK4A) and p53 are elevated during premature senescence induced by depletion of E2FBP1, and the depletion of p16(INK4A), but not p53 rescued senescent cells from growth arrest. Therefore, the premature senescence induced by E2FBP1 depletion is achieved through the p16(INK4A)-Rb pathway. Similar to human normal fibroblasts, the growth inhibition induced by E2FBP1 depletion is also observed in human tumor cells with intact p16(INK4A) and Rb. These results suggest that E2FBP1 functions as a critical antagonist to the p16(INK4A)-Rb tumor suppressor machinery by regulating PML stability.
Cell Line, Tumor ; Cells, Cultured ; Cellular Senescence ; genetics ; physiology ; Cyclin-Dependent Kinase Inhibitor p16 ; antagonists & inhibitors ; genetics ; physiology ; DNA-Binding Proteins ; deficiency ; genetics ; physiology ; Down-Regulation ; Fibroblasts ; Gene Expression Regulation ; Humans ; Intranuclear Inclusion Bodies ; metabolism ; MAP Kinase Signaling System ; Nuclear Proteins ; genetics ; metabolism ; physiology ; Promyelocytic Leukemia Protein ; Protein Isoforms ; Protein Stability ; RNA Interference ; Retinoblastoma Protein ; antagonists & inhibitors ; genetics ; physiology ; Transcription Factors ; deficiency ; genetics ; metabolism ; physiology ; Transfection ; Tumor Suppressor Protein p53 ; physiology ; Tumor Suppressor Proteins ; genetics ; metabolism ; physiology ; Ubiquitination ; ras Proteins ; metabolism

OBJECTIVE: To study the clinical significance of the expression of p53, p63 and p73 protein and the correlation of p53 and p63 in nasal and paranasal sinus carcinoma (NPSC). METHOD: The immunohistochemical technique was used to detect the expression of p53, p63 and p73 in 67 cases of NPSC, para-cancer tissues and non-cancer tissues. RESULT: The positive rate of p53 and p63 protein in NPSC was significantly higher than that in park cancer tissues and non-cancer tissues (P<0.01), and there were a positive correlation between the expression of p53 and p63 protein in NPSC (P<0.01). But the expression of p73 had no significant difference among NPSC, para-cancer tissues and non-cancer tissues (P>0.05). CONCLUSION There are positive correlation between p53 and p63 protein in NPSC, and they may play an important role in the tumorgenesis of NPSC. The p73 may not be associated with tumorgenesis of NPSC.
Carcinoma, Squamous Cell ; metabolism ; pathology ; DNA-Binding Proteins ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Nose Neoplasms ; metabolism ; pathology ; Nuclear Proteins ; metabolism ; Paranasal Sinus Neoplasms ; metabolism ; parasitology ; Trans-Activators ; metabolism ; Transcription Factors ; Tumor Protein p73 ; Tumor Suppressor Protein p53 ; metabolism ; Tumor Suppressor Proteins ; metabolism

Nasopharyngeal carcinoma (NPC) is a malignancy with remarkable ethnic and geographic distribution in southern China and Southeast Asia. Alternative to genetic changes, aberrant epigenetic events disrupt multiple genes involved in cell signaling pathways through DNA methylation of promoter CpG islands and/or histone modifications. These epigenetic alterations grant cell growth advantage and contribute to the initiation and progression of NPC. In this review, we summarize the epigenetic deregulation of cell signaling in NPC tumorigenesis and highlight the importance of identifying epigenetic cell signaling regulators in NPC research. Developing pharmacologic strategies to reverse the epigenetic-silencing of cell signaling regulators might thus be useful to NPC prevention and therapy.
Adaptor Proteins, Signal Transducing ; genetics ; metabolism ; Apoptosis ; genetics ; Carcinoma ; Cell Cycle ; genetics ; CpG Islands ; genetics ; DNA Damage ; genetics ; DNA Methylation ; Epigenesis, Genetic ; GTP Phosphohydrolases ; genetics ; metabolism ; Gene Silencing ; Humans ; Nasopharyngeal Neoplasms ; genetics ; metabolism ; pathology ; Repressor Proteins ; genetics ; metabolism ; Signal Transduction ; Tumor Suppressor Protein p53 ; genetics ; metabolism ; beta Catenin ; genetics ; metabolism ; ras Proteins ; genetics ; metabolism ; rho GTP-Binding Proteins ; genetics ; metabolism

<b>OBJECTIVEb>To explore the influence of BCL6, MYC, P53 genes abnormalities can on the prognosis of diffuse large B-cell lymphoma (DLBCL), and to identify independent prognostic factors for DLBCL in order to facilitate clinical prognosis and selection of stratification treatment for the patients.

<b>METHODSb>Sixty five newly diagnosed DLBCL pathological specimens were collected from 2009 to 2012. Interphase fluorescence in situ hybridization technique (I-FISH) was used to detect the status of BCL6, MYC and P53 genes. Clinical factors were combined with immunohistochemical results for multiple-factor survival analysis.

<b>RESULTSb>The rates of BCL6 gene rearrangement, P53 gene deletion and MYC rearrangement were 21.5% (14/65), 35.4% (23/65) and 7.7% (5/65), respectively. BCL6 rearrangement group has obviously poorer overall survival (OS)(P< 0.05). COX proportional hazards model analysis showed that gender, BCL6 protein, BCL6 rearrangement, Ki67 index were prognosis factors independent of international prognostic index (IPI).

<b>CONCLUSIONb>BCL6 can influence the prognosis of patients with DLBCL at gene and protein levels and both are independent prognostic factors for DLBCL.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; DNA-Binding Proteins ; genetics ; Female ; Gene Deletion ; Gene Rearrangement ; Humans ; In Situ Hybridization, Fluorescence ; Lymphoma, Large B-Cell, Diffuse ; diagnosis ; genetics ; mortality ; Male ; Middle Aged ; Mutation ; Prognosis ; Proto-Oncogene Proteins c-bcl-6 ; Proto-Oncogene Proteins c-myc ; genetics ; Survival Analysis ; Survival Rate ; Tumor Suppressor Protein p53 ; genetics ; Young Adult