Animals ; Homeodomain Proteins ; metabolism ; Humans ; Skin ; metabolism ; Wound Healing ; physiology

Formaldehyde (FA) is a ubiquitous toxic organic compound, and it has been regarded as a leukemogen. However, the mechanisms by which FA induces bone marrow toxicity remain unclear. The present study was aimed to examine the bone marrow toxicity caused by FA and the mechanism involving the expression changes of peroxiredoxin3 (Prx3) in this process. The mice were divided into four groups with 6 mice per group. Animals in the control group were exposed to ambient air and those in the FA groups to different concentrations of FA (20, 40, 80 mg/m(3)) for 15 days in the separate inhalation chambers, 2 h a day. At the end of the 15-day experimental period, all mice were killed. Bone marrow cells were obtained. The level of hydrogen peroxide (H2O2), the apoptosis rate, and the activities and protein expression levels of caspase-3 and caspase-9 were determined by biochemical assay, flow cytometry and immunohistochemistry, respectively; DNA damage and Prx3 expression levels were measured by single cell gel eletrophoresis immunohistochemistry and Western blotting, respectively. The results showed that the H2O2 level and cell apoptosis rate were significantly increased in FA groups relative to the control group. Caspase-3 and caspase-9 activities and their protein expression levels were markedly increased as well. Additionally, FA also increased the rate of DNA damage and the expression level of Prx3 compared with control group. Our study suggested that a certain concentration of FA causes the bone marrow toxicity by regulating the expression of Prx3.
Animals ; Blotting, Western ; Bone Marrow ; drug effects ; metabolism ; Formaldehyde ; pharmacology ; Homeodomain Proteins ; metabolism ; Male ; Mice

As one member of the HOX gene family, HOXB4 gene is a specific transcription factor, playing an important role in regulating balance between self-renewal and differentiation of hematopoietic stem/ progenitor cells (HSPC). Recombinant human TAT-HOXB4 protein carrying the protein transduction domain of the HIV transactivating protein (TAT) is comparable to that induced by the human HOXB4 retrovirus. TAT-HOXB4-expanded HSPC populations keep the potential of differentiation and long-term repopulation in vivo. In this article, whether TAT-HOXB4 protein can induce leukemia or can be applied to clinical treatment are reviewed, summarizing the structures of HOX gene and HOXB4 gene, role of HOXB4 in HSPC proliferation and different ion, HOX gene and leukemia, TAT-HOXB4 protein, Clinical application prospect of TAT-HOXB4 protein and so on.
Cell Culture Techniques ; Cell Differentiation ; Hematopoietic Stem Cells ; cytology ; Homeodomain Proteins ; metabolism ; Humans ; Recombinant Proteins ; Transcription Factors ; metabolism

BACKGROUND: CDX1 and CDX2 are members of the caudal-type homeobox gene family and control the proliferation and differentiation of intestinal mucosal cells. Their expressions are commonly reduced in colorectal cancer, but reports about the relationships between their expressions and clinicopathologic features are rare. The aim of this study was to examine the expressions of CDX1 and CDX2 mRNAs in colorectal cancers and to assess the relationships between their expressions and clinicopathologic features. METHODS: CDX1 and CDX2 mRNA expressions were analyzed by real-time polymerase chain reaction in 48 colorectal cancers and in adjacent non-tumorous normal mucosal tissue. RESULTS: CDX1 and CDX2 mRNA expressions were significantly reduced in colorectal cancer tissues versus normal mucosal tissues (p=0.001, p=0.042, respectively). As compared with paired normal mucosal tissues, colorectal tissues showed reduced CDX1 mRNA expression in 64.6% (31/48) and reduced CDX2 mRNA expression in 66.7% (32/48) of cases. A statistically significant positive correlation was found between the expressions of CDX1 mRNA and CDX2 mRNA in colorectal cancer (r=0.543, p< 0.001). However, the expressions of CDX1 and CDX2 mRNAs were not related to age, sex, cancer location, differentiation, lymphatic or vascular invasion, lymph node metastasis, stage or serum carcinoembryonic antigen level. CONCLUSIONS: CDX1 and CDX2 mRNA expressions were found to be significantly reduced in colorectal cancers, but these expressional changes were not found to be related to clinicopathologic features.
RNA, Messenger/*metabolism ; Polymerase Chain Reaction ; Middle Aged ; Male ; Humans ; Homeodomain Proteins/*metabolism ; Female ; Colorectal Neoplasms/*metabolism

OBJECTIVETo investigate the expression of HOXA13 gene in stage-II(a esophageal squamous cell carcinoma(ESCC), and to evaluate its relationship with clinicopathological characteristics and prognosis.

METHODSThe expression of HOXA13 was examined by immunohistochemistry(IHC) in specimens from 39 patients with ESCC of stage-II(a, who underwent resection from 1995 to 2002. SPSS software was used to analyze the relationship between HOXA13 expression and clinicopathological characteristics and prognosis of patients.

RESULTSThe expression of HOXA13 protein was detected in ESCC tissue, and the positive rate was 61.5%. The median survival time of patients without HOXA13 expression(>72 months) was significantly longer than those with HOXA13 expression (24 months)( P=0.023). Multivariate analysis showed that HOXA13 expression was independent predictor of disease-free survival time of patients with ESCC.

CONCLUSIONThe expression of HOXA13 can be detected in ESCC and is a negative independent predictor of disease-free survival, which implies that HOXA13 might play a role in ESSC, and may be used as a clinical tumor marker of ESCC.

Carcinoma, Squamous Cell ; metabolism ; pathology ; Esophageal Neoplasms ; metabolism ; pathology ; Female ; Homeodomain Proteins ; metabolism ; Humans ; Male ; Middle Aged ; Neoplasm Staging ; Prognosis

OBJECTIVETo introduce the eukaryotic expression vector pEGFP-C1-PDX-1 into nestin-positive cell derived from bone marrow stromal cells by nucleofection and optimize the conditions for transfection.

METHODSThe recombinant plasmid was transfected into bone marrow stromal cells-derived nestin-positive cells with varied DNA quantities or the serum concentration in the medium. The expression of PDX-1 gene in the transfected cells was detected by RT-PCR.

RESULTSSatisfactory efficiency of transfection was achieved with the DNA quantity of 2-10 microg and medium serum concentration of 20%. PDX-1 expression was detected in the transfected cells by RT-PCR.

CONCLUSIONThe optimized transfection conditions result in enhanced efficiency of PDX-1 gene transfection into nestin-positive cells derived from bone marrow stromal cells, which may serve as the seed cells in tissue-engineering.

Bone Marrow Cells ; metabolism ; Genetic Vectors ; Homeodomain Proteins ; genetics ; Humans ; Intermediate Filament Proteins ; metabolism ; Nerve Tissue Proteins ; metabolism ; Nestin ; Plasmids ; Stromal Cells ; metabolism ; Trans-Activators ; genetics ; Transfection ; methods

OBJECTIVETo investigate the expressions of Engrailed-2 (EN2) and β-catenin in bladder urothelial carcinoma and explore their significance.

METHODSSixty bladder urothelial carcinoma samples of different grades and stages and 10 normal bladder mucosal tissues were examined for expressions of EN2 and β-catenin proteins and mRNA using immunochemistry, Western blotting and RT-PCR. RESULTS Compared to normal bladder mucosa, bladder urothelial carcinoma tissues showed significantly increased expressions of EN2 and β-catenin proteins (P<0.05), and the high-grade carcinoma tissues exhibited significantly stronger expressions than the low-grade ones (P<0.05); the expressions of the proteins increased also significantly with advanced pathological stages of bladder urothelial carcinoma (P<0.05). The expressions of EN2 and β-catenin mRNAs showed a consistent pattern of changes with their protein expressions.

CONCLUSIONThe expressions of EN2 and β-catenin are significantly increased in bladder urothelial carcinoma. EN2 may contribute to the development and progression of bladder urothelial carcinoma by activating Wnt/β-catenin signal pathway.

Adult ; Aged ; Aged, 80 and over ; Carcinoma ; metabolism ; pathology ; Female ; Homeodomain Proteins ; metabolism ; Humans ; Male ; Middle Aged ; Nerve Tissue Proteins ; metabolism ; Urinary Bladder Neoplasms ; metabolism ; pathology ; beta Catenin ; metabolism

OBJECTIVETo investigate the function of AT motif binding factor 1 (ATBF1) in colorectal cancer.

METHDSATBF1 protein expression was detected in 146 pairs of colorectal cancer tissues and the adjacent tissues using immunohistochemistry. ATBF1 protein expression was also examined in colorectal cell lines with laser confocal microscopy. ATBF1-A protein expression in colorectal cancer tissues of different differentiation grades and in the colorectal cancer cell lines were detected with Western blotting. The expressions of ATBF1 mRNA in 38 moderately differentiated colorectal cancer tissues and the paired adjacent tissues and in the colorectal cancer cell lines were tested using RT-PCR.

RESULTSATBF1 protein expression levels in colorectal cancer tissues and adjacent tissues differed significantly (P<0.001), and its expression increased significantly in positive correlation with the grade of tumor differentiation (P<0.001) but in negative correlation with tumor metastasis. ATBF1 mRNA expression and ATBF1 protein expression were significantly correlated (P=0.100). The ATBF1 protein and mRNA expressions also differed significantly among different colorectal cancer cell lines.

CONCLUSIONATBF1 executes the role of a tumor suppressor gene in colorectal cancer, and its protein expression is associated with tumor differentiation and lymph node metastases.

Blotting, Western ; Cell Differentiation ; Colorectal Neoplasms ; metabolism ; Genes, Tumor Suppressor ; Homeodomain Proteins ; metabolism ; Humans ; Immunohistochemistry ; Lymphatic Metastasis ; RNA, Messenger

HOXA10 gene plays an essential role in differentiation of the endometrium and in human reproduction. The aim of this study was to investigate the regulatory effect of sex steroids and HB-EGF on HOXA10 gene in Ishikawa cells. Ishikawa cells were incubated with 17-beta estradiol (10(-8) mol/L), medroxyprogesterone acetate (MPA) (10(-6) mol/L), RU486 (10(-5) mol/L) or HB-EGF (10 ng/mL) for 48 h respectively. The expression of HOXA10 gene was detected by immunofluorescence, reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting. Our results showed that either estrogen alone, progestin alone or progestin combined with estrogen could significantly increase the expression of HOXA10 gene 48 h after the treatment (P<0.05). But estrogen combined with progestin and RU486 could inhibit the up-regulation by estrogen and progestin. HB-EGF could elevate the expression of HOXA10 gene 48 h after the treatment (P<0.05). It is concluded that both estrogen and progestin can up-regulate the expression of HOXA10 gene in Ishikawa cells, but RU486 can inhibit the effect and HB-EGF can elevate the expression level of HOXA10 gene.
Cell Line, Tumor ; Endometrial Neoplasms/*pathology ; Estradiol/*pharmacology ; Genes, Homeobox ; Homeodomain Proteins/genetics ; Homeodomain Proteins/*metabolism ; Intercellular Signaling Peptides and Proteins/*pharmacology ; Medroxyprogesterone Acetate/*pharmacology ; RNA, Messenger/genetics ; RNA, Messenger/metabolism

Pdx-1, an important transcription factor highlighting in the early pancreatic development, islet functions and pancreatic disorders, needs to be more investigated in zebrafish, and siRNA is still seldom applied in zebrafish embryo-related research. Our aim was to explore the role of pdx-1 in pancreatic development of zebrafish embryos by using siRNA approach. Microinjection, reverse transcriptase-PCR (RT-PCR), in situ hybridization and immunofluorescent staining were used in this research, and the morphology of the islet in normal zebrafish embryos, and in those treated with the siRNA specific to pdx-1 (siPDX-1) or siGFP was observed and compared. The expression of pdx-1 was detected in the stages of 1-cell, 2-cell, 4-cell, 8-cell, 16-cell, 16-hour by RT-PCT. The in situ hybridization and immunofluorescent staining results showed that siPDX-1 disturbed the formation of the islet in zebrafish embryos. Pdx-1 played multiple roles in maintaining the phenotype of the islet during embryogenesis in zebrafish.
Embryo, Nonmammalian ; Homeodomain Proteins/genetics ; Homeodomain Proteins/*metabolism ; Islets of Langerhans/cytology ; Islets of Langerhans/*embryology ; Islets of Langerhans/metabolism ; RNA Interference ; RNA, Small Interfering/*genetics ; Trans-Activators/genetics ; Trans-Activators/*metabolism ; Zebrafish