TGF-beta 1 controls the expression of numerous genes, including early response and cellular matrix genes. However, signal-transducing mechanism underlying this regulation gene is not fully understood. In study, we investigated whether redox plays a role in signal transduction mouse osteoblastic cell line (MC3T3-E1). The overall intracellular oxidized state cells, when measured using 2',7'-dichlorofluorescin diacetate by laser-scanning confocal microscopy, was increased transiently after...

H 2 O , like other oxidants, is known to act as a mitogen at low concentrations in resting Balb/3T3 or mouse epidermal JB6 cells. We described previously that induces some early response genes extended these observations using another cell line, MC3T3 (mouse osteoblastic) cells by examination of transcriptional activity and inhibitors protein kinases. increased the expressions c‐ fos jun, egr ‐1 JE which are be induced mitogenic stimuli many types Exogenous addition mRNA levels genes,...

Transforming growth factor(TGF)β1 is a potent inhibitor of in mouse osteoblastic MC3T3‐E1 cells. To isolate genes that are induced by TGFβ1, the differential screening method was adopted using cDNA library constructed from cells treated with TGFβ1 for 4 h. Six independent clones were isolated (TGFβ‐stimulated clone, TSC‐5, TSC‐36, TSC‐115, TSC‐128, TSC‐160 and TSC‐161), expression which increased TGFβ1‐treatment maximal at 6–10 The steady‐state levels TSC‐128 almost tenfold, whereas those...

Transforming growth factor (TGF) beta 1 is a potent cytokine that inhibits the of several types cells. Our earlier study suggested mouse osteoblastic cell line, MC3T3-E1, was sensitive to inhibition by TGF and this effect partly mediated H2O2. To identify molecules participate in negative regulation these stimuli, we carried out differential screening cDNA libraries isolated set genes induced 1. Among clones isolated, one originally named tsc-5 found be H2O2 as well Analysis renamed hic...

Abstract Although the causal relationship between chronic inflammation and carcinogenesis has long been discussed, molecular basis of relation is poorly understood. In present study, we focused on reactive oxygen species (ROS) their signals under inflammatory conditions leading to epithelial cells found that repeated treatment with a low dose H2O2 (0.2 mmol/L) for periods 2 4 days caused phenotypic conversion mouse NMuMG mammary from fibroblast-like as in malignant transformation. The...

A gene sequence (TSC-22) that is induced by transforming growth factor (TGF) 81 was isolated differential screening of a XgtlO cDNA library constructed from poly(A)+ RNA mouse osteoblastic cells treated with TGF-81 for 2 h.TSC-22 expression transcriptionally activated TGF-81.It also phorbol 12-myristate 13-acetate, serum, cholera toxin, or dexamethasone, but not appreciably epidermal factor.Its induction rapid and transient, reaching peak h after treatment, resistant to cycloheximide like...

We examined the role of superoxide in increase intracellular pH (pHi) human histiocytic leukemia U937 cells treated with 4 beta-phorbol-12,13-didecanoate (4 beta-PDD) or serum. beta-PDD serum induced a rapid pHi, and antioxidants such as dismutase (SOD), vitamin E, butylated hydroxyanisole (BHA) were found to inhibit amiloride-sensitive pHi by beta-PDD. SOD inhibited caused serum, essentially same was concanavalin A-stimulated mouse thymocytes. Also, superoxide-generating system,...

The role of the electron transport chain (ETC) in cell proliferation control beyond its crucial function supporting ATP generation has recently emerged. In this study, we found that, among four ETC complexes, complex I (CI)‐mediated NAD + regeneration is important for cancer proliferation. cells, a decrease CI activity by RNA interference (RNAi) against NADH:ubiquinone oxidoreductase core subunit V1 ( NDUFV1 ) arrested cycle at G 1 /S phase, accompanying upregulation p21 Cip1...

The hic-5 gene encodes a novel protein with Zn finger-like (LIM) motifs, the expression of which increases during cellular senescence. ectopic in nontumorigenic immortalized human fibroblasts, whose levels were significantly reduced comparison those mortal cells, decreased colony-forming efficiency. Stable clones expressing high mRNA showed higher mRNAs for several extracellular matrix-related proteins, along alteration an alternative splicing as seen senescent cells and c-fos inducibility....

Hic-5 is a paxillin homologue that localized to focal adhesion complexes.Hic-5 and share structural homology interacting factors such as kinase (FAK), Pyk2/CAK␤/RAFTK, PTP-PEST.Here, we showed inhibits integrin-mediated cell spreading on fibronectin in competitive manner with NIH 3T3 cells.The overexpression of sequestered FAK from paxillin, reduced tyrosine phosphorylation FAK, prevented paxillin-Crk complex formation.In addition, Hic-5-mediated inhibition was not observed mouse embryo...

hic-5 was originally isolated as an H(2)O(2)-inducible cDNA clone whose product normally found at focal adhesions. In this study, we that Hic-5 accumulated in the nucleus response to oxidants such H(2)O(2). Other adhesion proteins including paxillin, most homologous Hic-5, remained cytoplasm. Mutation analyses revealed C- and N-terminal halves of contributed its nuclear localization a positive negative manner, respectively. After finding leptomycin B (LMB), inhibitor export signal (NES),...

The Hic-5 protein is encoded by a transforming growth factor-β1- and hydrogen peroxide-inducible gene,hic-5, has striking similarity to paxillin, especially in their C-terminal LIM domains. Like localized focal adhesion plaques association with kinase cultured fibroblasts. We carried out yeast two-hybrid screening identify cellular factors that form complex using its domains as bait, we identified cytoplasmic tyrosine phosphatase (PTP-PEST) one of the partners Hic-5. These two proteins are...

Hydrogen peroxide-inducible clone (Hic)-5 is induced during the senescent process in human fibroblasts, and overexpression of Hic-5 induces a senescence-like phenotype. Structurally, paxillin, 68-kDa cytoskeletal protein, share homology such as LD motifs N-terminal half LIM domains C-terminal half. Here we show that binds to focal adhesion kinase (FAK) by its domain, localized adhesions domains. However, not tyrosine phosphorylated either coexpressed FAK COS cells or integrin stimulation...

Abstract Anchorage dependence of cellular growth and survival prevents inappropriate cell or in ectopic environments, serves as a potential barrier to metastasis cancer cells. Therefore, obtaining better understanding anchorage-dependent responses normal cells is the first step understand impede anchorage independence finally eradicate during metastasis. Anoikis, type apoptosis specifically induced by lack appropriate cell-extracellular matrix adhesion, has been established dominant response...

Two of mitogen-activated protein kinases (MAPK), p44mapk/p42mapk extracellular signal-regulated (ERK1/2), translocate into nuclei following activation and play critical roles in connecting the signal to gene expression allowing cell-cycle entry. Here we found that nuclear translocation ERK1/2 response growth stimuli was significantly inhibited senescent cells were irreversibly arrested, compared with presenescent cells. The step these enzymes not impaired, since phosphorylated activated as...

Abstract Hic‐5 is a LIM protein with striking similarity to paxillin, and shuttles between focal adhesions the nucleus. Our previous study suggested that participates in transcriptional control of several genes such as c‐ fos p21 genes. In present study, we examined function nucleus using promoter region gene. When localized nucleus, was found transactivate through two five Sp1 sites proximal TATA box. The effect mediated by transactivation domain functional interaction p300 LIM4 domain....

Hic-5 is a focal adhesion protein belonging to the paxillin LIM family that shuttles in and out of nucleus. In present study, we examined expression among mouse tissues by immunohistochemistry found its only smooth-muscle cells several tissues. This result consistent with previous report on adult human contradicts relatively ubiquitous paxillin, most homologous Hic-5. One factor characterizing vivo continuous exposure mechanical stretching organs. To study involvement cellular responses...

Recent evidence suggests that bone marrow-derived cells may contribute to repair and lesion formation following vascular injury. In most studies, were tracked by transplanting exogenous into marrow had been compromised irradiation. It remains be determined whether endogenous circulating progenitors actually arterial remodeling under physiological conditions. Here, we established a parabiotic model in which two mice conjoined subcutaneously without any anastomosis. When wild-type joined with...

Mitochondria are considered to play an important role in oxidative stress response since they a source of reactive oxygen species and also targeted by these species. This study examined the mitochondrial conditions cells epithelial origin that were exposed H2O2 found decline membrane potential along with specific loss UQCRC1, sub-unit complex III, suggesting dysfunction occurs upon exposure stress. observation led hypothesis certain cellular responses occurred because dysfunction. When...