Biomedical subjects

S Okabe

Publications and source records attributed to S Okabe.

At least 127 records · Page 7Linked to original sources

PubMed1999

Epidermal growth factor protects rat epithelial cells against acid-induced damage through the activation of Na+/H+ exchangers.

We examined the effect of epidermal growth factor (EGF) on acid-induced cell damage in rat gastric epithelial cells (RGM1) and investigated the mechanisms of this effect. Cells were incubated with EGF for 5, 15, 30, and 60 min, and then immersed in an acidified medium (pH 4.0) for 30 min to induce cell damage. EGF prevented cell damage in a concentration- and time-dependent manner. EGF reduced the effects of the acidified medium on the cells, preventing the reduction of intracellular pH. Replacement of Na+ with K+ in the acidified medium canceled the effect of EGF. Indomethacin and W-7 (a Ca-calmodulin inhibitor) did not alter the protective effect of EGF. In contrast, genistein (a tyrosine kinase inhibitor), amiloride (a Na+/H+ exchangers I and II inhibitor), and wortmannin (a phosphatidylinositol 3-kinase inhibitor) significantly decreased the effect of EGF. Expression of Na+/H+ exchangers type I and type II was confirmed by reverse transcription-polymerase chain reaction. Our results demonstrate that EGF prevents acid-induced cell damage, most likely through the activation of a Na+/H+ exchanger II via phosphatidylinositol 3-kinase.

Animals↗

PubMed1998

Anticancer activity of morphine and its synthetic derivative, KT-90, mediated through apoptosis and inhibition of NF-kappaB activation.

We recently reported that morphine inhibits growth of various human cancer cell lines (IC50/2.7-8.8 mM). We then extended the study using newly synthesized morphine derivatives, such as KT-90 and KT-87, which are analgesics 5 times more potent than morphine. KT-90 was found to inhibit growth of human cancer cell lines (IC50/42-70 microM) up to 80 times more potently than morphine. As for mechanisms of action, KT-90 and morphine induced apoptosis, and inhibited tumor necrosis factor alpha (TNF-alpha) gene expression induced by tumor promoters, okadaic acid and 12-O-tetradecanoylphorbol-13-acetate, associated with reduction of NF-kappaB DNA binding activity. This paper provides evidence that KT compounds confirmed the presence of anticancer activity of morphine in addition to its analgesic action.

Analgesics↗

PubMed1998

Genomic organization and mutation analyses of the DR5/TRAIL receptor 2 gene in colorectal carcinomas.

The DR5/TRAIL receptor 2 gene is a novel TNF receptor family member and induces apoptosis by overexpression of its product. DR5 is located on chromosome 8p21, where allelic deletions are often observed in advanced colorectal cancers. This evidence led us to examine whether or not mutations of the DR5 gene exist in these tumors. We determined the genomic structure of DR5, which consists of nine exons. DR5 has two alternatively spliced isoforms. All seven colorectal cancer (CRC) cell lines examined expressed the longer isoform predominantly. No somatic mutation was found in 41 CRC cases. Loss of heterozygosity of DR5 was found in nine (52.9%) of the 17 informative cases, which is similar to the previously reported frequencies observed for 8p21-22 in colorectal cancers. These findings suggest that inactivation of the DR5 gene may play only a small role, if any, in colorectal tumorigenesis.

Apoptosis↗

PubMed1998

Allelotype analysis of early colorectal cancers with lymph node metastasis.

Several studies have indicated that frequent allelic losses in some specific chromosomal regions occur during colorectal cancer (CRC) progression. To clarify the correlation between such allelic losses and metastatic potential, the allelotype of lymph node-positive early CRCs, which are small but extremely malignant cancers consisting of metastatically competent cells, were investigated. Nineteen paraffin-embedded specimens of early CRC (pT1 tumors according to TNM classification) with positive lymph nodes were collected. The tumor tissues were examined for loss of heterozygosity (LOH), using microsatellite markers on chromosomes 1p34-36, 8p21-22, 14q32, 18q21 and 22q12-13. The relationship between p53 protein expression and the metastatic status was also investigated by immunohistochemical staining. A group of 20 early CRCs with negative lymph nodes having a similar distribution of macroscopic appearance were used as controls. Among the 19 node-positive tumors, LOH at 8p21-22 and 18q21 was detected in 11 cases (57.9%) and 17 cases (89.4%), respectively. Allelic losses within these 2 regions in node-positive tumors were significantly more frequent than that in node-negative ones (p < 0.01). No significant correlation was found between LOH at 1p34-36, 14q32 or 22q12-13 and lymph node metastasis. p53 protein expression was not significantly associated with lymph node metastasis. Our results suggest that putative tumor suppressor genes, which may be involved in the metastatic process of CRC, are located on chromosomes 8p21-22 and 18q21. Allelic losses in these regions are possible risk factors for lymph node metastasis of early CRC.

Adult↗

PubMed1998

Cancer inhibition by green tea.

Green tea is now an acknowledged cancer preventive in Japan. This paper discusses several important features of (-)-epigallocatechin gallate (EGCG), the main constituent of green tea and tea polyphenols. EGCG and other tea polyphenols inhibited growth of human lung cancer cell line, PC-9 cells with G2/M arrest. 3H-EGCG administered by p.o. intubation into mouse stomach revealed that small amounts of 3H-activity were found in various organs where EGCG and green tea extract had previously demonstrated their anticarcinogenic effects, such as skin, stomach, duodenum, colon, liver, lung and pancreas. Cancer onset of patients who had consumed over 10 cups of green tea per day was 8.7 years later among females and 3.0 years later among males, compared with patients who had consumed under three cups per day. The mechanisms of action of EGCG were briefly discussed with regard to inhibition of tumor necrosis factor-alpha (TNF-alpha) release.

3T3 Cells↗

PubMed1998

Hippocampal synaptic plasticity in mice overexpressing an embryonic subunit of the NMDA receptor.

The effects of changing NMDA receptor subunit composition on synaptic plasticity in the hippocampus were analyzed by creating transgenic mice overexpressing NR2D, a predominantly embryonic NMDA receptor subunit. NMDA-evoked currents in the transgenic mice had smaller amplitudes and slower kinetics. The transgenics also displayed age-dependent deficits in synaptic plasticity in area CA1 of the hippocampus. Long-term depression was selectively impaired in juvenile mice when NR2D overexpression was moderate. In mature mice, overexpression of NR2D was associated with a reduction of both NR2B and Ca2+-independent activity of Ca2+- and calmodulin-dependent protein kinase II. These biochemical changes were correlated with a marked impairment of NMDA-dependent long-term potentiation, but spatial behavior was normal in these mice. These results show that the developmental regulation of NMDA receptor subunit composition alters the frequency at which modification of synaptic responses occur after afferent stimulation.

2-Amino-5-phosphonovalerate↗

PubMed1998

Genomic structure of the human Smad3 gene and its infrequent alterations in colorectal cancers.

The Smad3 gene is a member of the Smad family, vertebrate homologues of Drosophila Mad, and its gene product is a cytoplasmic element in the TGF-beta signaling pathway. Smad2 and Smad4/DPC4, other members of the Smad family, are possibly tumor suppressor genes because alterations of these genes occurred in various carcinomas. We determined the genomic structure of human Smad3 which consists of nine exons. Then we examined whether or not Smad3 gene mutations exist in sporadic and hereditary non-polyposis colorectal cancers and found no mutations in the entire coding region in 50 cancers. Loss of heterozygosity of Smad3 was observed in two of the 17 (11.8%) informative cases using a polymorphism found in intron 2. These findings suggest that the Smad3 gene may not play an important role in the tumorigenesis of colorectal cancers.

Base Sequence↗

PubMed1998

Molecular pathogenesis of interstitial pneumonitis with TNF-alpha transgenic mice.

Tumour necrosis factor alpha (TNF-alpha) transgenic mice, which overexpress TNF-alpha in the lungs, develop interstitial pneumonitis resembling idiopathic pulmonary fibrosis (IPF) in humans. Transgenic mice were used to study molecular pathogenesis of interstitial pneumonitis with regard to sequential histological changes and cytokine network induced by TNF-alpha. The authors divided the histological process of interstitial pneumonitis into three stages: early stage with lymphocytic infiltration in alveolar septa, middle stage with recruitment of macrophages, and late stage with hyperplasia of epithelial cells and mild fibrosis. As for cytokine network, prolonged overexpression of TNF-alpha along with increasing interleukin 6 (IL-6) were associated with the progression of interstitial pneumonitis. Increasing IL-1 was found only in the early stage, the beginning of lymphocyte proliferation. The mRNA level of an anti-inflammatory cytokine, IL-10, was constantly enhanced in the lungs of transgenic mice. However, transforming growth factor beta 1 (TGF-beta 1) protein decreased, which is closely associated with prolonged TNF-alpha synthesis, resulting in development of chronic inflammation and less severe fibrosis in the lungs of this animal model, analogous to inflammatory stage of human IPF. TNF-alpha transgenic mice enabled the analysis of the sequential process of interstitial pneumonitis as a model of IPF pathogenesis in humans, the results of which will give rise to new therapeutic measures for human IPF.

Animals↗

PubMed1998

A clinicopathological investigation on superficial early invasive carcinomas of the colon and rectum.

Due to recent advances in endoscopic surgical techniques, it has now become possible to perform endoscopic resection of most early invasive carcinomas of the colon and rectum (EIC) even if the lesions have invaded the submucosa. In the present study, we investigated the microscopic characteristics of superficial EIC compared with protruding-type EIC, focusing particular attention on histological type, the presence or absence of vascular invasion, the extent of submucosal invasion, and other adverse prognostic factors, to establish appropriate treatment strategies. Our findings led us to conclude that: (1) most cases of EIC can be cured by endoscopic resection if their gross aspects are classified as type IIc, superficial depressed, or type IIa, superficial elevated; (2) colorectal resection with lymph node dissection should be performed first for type IIa + IIc EIC because these lesions are apt to be associated with a large number of adverse prognostic factors; (3) subsequent colorectal resection should be performed after initial endoscopic treatment of EIC if there are adverse prognostic indicators of metastasis in the endoscopically resected specimen, such as moderately or poorly differentiated adenocarcinoma, lymphatic invasion, venous invasion, or extensive submucosal invasion.

Adenocarcinoma↗

PubMed1998

Pathological changes in the formation of Helicobacter pylori-induced gastric lesions in Mongolian gerbils.

We examined pathological changes in the formation of Helicobacter pylori-induced gastric lesions in Mongorian gerbils. H. pylori (NCTC11637) was orally administered once to the animals and was detected in the gastric mucosa of all gerbils given the bacteria. The number of viable H. pylori increased during the initial two weeks and thereafter reached a plateau level. The initial pathological changes were found at one week, ie, edema/congestion and a white viscous substance only in the antrum. At two weeks, superficial damage appeared in the antrum, although inflammatory cell infiltration had not occurred. Gastritis with lymphoid follicles was observed in the antrum and fundus from three weeks. At four weeks, mucosal lesions were detected as a few hemorrhagic spots in the fundus adjacent to the antrum. In the control animals, however, no pathological changes were observed even at four weeks. In the gastric mucosa infected with H. pylori, myeloperoxidase activity was negligible at two weeks, but was extremely elevated at four weeks. Similarly, neutrophil chemotactic activity was only slightly increased at two weeks, but was markedly elevated at four weeks. These results indicate that H. pylori infection induces initial pathological changes only in the antrum, but mucosal lesions occur in the fundus adjacent to the antrum. Furthermore, it is demonstrated that the initial superficial damage is generated by factors other than chemokines and neutrophil-associated factors, although mucosal inflammation may contribute to the subsequent formation of lesions and ulcers.

Animals↗

PubMed1998

Interleukin-1beta inhibits growth factor-stimulated restoration of wounded rat gastric epithelial cell monolayers.

We examined the effect of interleukin-1beta (IL-1beta) on spontaneous and enhanced restoration (cell migration and proliferation) using an in vitro wound model comprising a confluent monolayer of rat gastric epithelial RGM1 cells. Repair of an artificial wound in a cell monolayer was found to be time- and concentration-dependent when the cells were incubated with epidermal growth factor (EGF) or transforming growth factor (TGF) -alpha alone for up to 24 hr. The growth factors also stimulated DNA synthesis significantly for 24 hr in a concentration-related manner. IL-1beta had no effect on wound restoration in the absence of the growth factors. However, it markedly inhibited the restoration enhanced by EGF and TGF-alpha, the inhibition being about 60% and 70%, respectively. In addition, IL-1beta significantly reduced the DNA synthesis stimulated by the growth factors. The EGF- and TGF-alpha-enhanced restoration was reduced by about 30% by mitomycin C, which potently inhibited the stimulated DNA synthesis. Mitomycin C had no effect on the spontaneous restoration. Even when treated with mitomycin C, the inhibitory effect of IL-1beta on the enhanced wound repair was still observed; however, the extent of the inhibition was decreased. These results indicate that IL-1beta inhibits the migration as well as the proliferation of gastric epithelial cells enhanced by EGF and TGF-alpha, resulting in a failure of wound healing.

Animals↗

PubMed1998

Effects of cytokines, without and with Helicobacter pylori components, on mucus secretion by cultured gastric epithelial cells.

Cytokines are suspected to play a crucial role in the pathogenesis of Helicobacter pylori-associated gastric diseases. Hence, considerable attention has been paid to the actions of cytokines on gastric cells. We examined the effects of cytokines on mucus secretion by gastric epithelial cells, without or with H. pylori components. Mucus secretion by cultured gastric epithelial cells was assessed as secretion of [3H]glucosamine-prelabeled high-molecular-weight glycoproteins. Interleukin (IL)-1beta and IL-6 significantly stimulated mucus secretion, but other cytokines such as IL-7, IL-8, IL-10, interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha had no effect. H. pylori lysate caused a decrease in both basal and stimulated secretion of mucus. In addition, IFN-gamma significantly potentiated the lysate-induced reduction of basal and stimulated secretion. Cell viability was not affected by any of treatments. These results indicate that IL-1beta and IL-6 stimulate mucus secretion, while IFN-gamma potentiates H. pylori-decreased secretion by gastric epithelial cells.

Animals↗

PubMed1998

Tumourigenicity of MTG8, a leukaemia-related gene, in concert with v-Ha-ras gene in BALB/3T3 cells.

The MTG8 (ETO) gene has been identified as the translocation partner of AML1 (PEBP2alphaB or CBFalpha2) gene in the AML1/MTG8 (ETO) fused gene caused by t(8;21) translocation in human acute myeloid leukaemia, M2 type. Although AML1/MTG8 chimaeric protein is known to inhibit the functioning of AML1 protein, the precise function of MTG8 gene itself is not known yet. We studied the significance of MTG8 gene in the oncogenicity of AML1/MTG8 fused gene, by introducing full-length MTG8 cDNA into both BALB/3T3 cells containing v-Ha-ras gene (Bhas 42 cells) and BALB/3T3 cells without v-Ha-ras gene. Irrespective of the overexpression of MTG8 gene in both groups of cells, Bhas-MTG8 clones which contained v-Ha-ras gene and expressed the MTG8 gene at a level more than twice that of parental Bhas 42 cells induced cell transformation, whereas BALB-MTG8 clones without v-Ha-ras gene did not. Furthermore, injection of the transformed Bhas-MTG8 clones into the subcutaneous tissue of nude mice induced tumours, whereas that of BALB-MTG8 clones did not. These results suggest that MTG8 gene product, in cooperation with viral Ras protein, resulted in tumour formation. We provide the first evidence that MTG8 gene by itself has a carcinogenic property within the AML1/MTG8 (ETO) fused gene.

3T3 Cells↗

PubMed1998

Survival and synaptogenesis of hippocampal neurons without NMDA receptor function in culture.

Physiological and morphological properties of cultured hippocampal neurons were measured to investigate whether NMDA receptors play a role in survival and differentiation. Neurons dissociated from mouse embryos with different NMDAR1 genotypes were grown in culture. Electrophysiological analysis verified the absence of NMDA receptor-mediated currents in neurons taken from homozygous mutant (NR1-/-) embryos. The number of surviving hippocampal neurons was 2.5-fold higher in cultures from the NR1-/- embryos compared with wild type (NR1 +/+) and heterozygous (NR1+/-) controls. Despite the lack of NMDA receptor function, NR1-/- neurons formed synapsin I-positive presynaptic boutons associated with MAP2ab-positive dendrites in culture. Confocal microscopic analysis of Dil labelled neurons confirmed the presence of dendritic spines on NR1-/- neurons with 80% of the density found in NR1 +/+ neurons. These results suggest that the NMDA receptor has little effect on general features of neuronal differentiation. In contrast, there is clear effect on neuronal survival. This finding establishes neuron number in standard culture conditions as a measure of NMDA receptor activity.

Animals↗

PubMed1998

Wide distribution of [3H](-)-epigallocatechin gallate, a cancer preventive tea polyphenol, in mouse tissue.

The increasing recognition of green tea and tea polyphenols as cancer preventives has created a need for a study of their bioavailability. For this purpose, we synthesized [3H] (-)-epigallocatechin gallate ([3H]EGCG) with a specific activity of 48.1 GBq/mmol and directly administered the solution into the stomachs of CD-1 female or male mice. Radioactivity in the digestive tract, various organs, blood, urine and feces was measured with an oxidizer at various times after administration and significant radioactivity was found in the previously reported target organs of EGCG and green tea extract (digestive tract, liver, lung, pancreas, mammary gland and skin), as well as other organs (brain, kidney, uterus and ovary and testes) in both sexes. Incorporation of radioactivity in the cells was confirmed by microautoradiography. Within 24 h, 6.6 (females) and 6.4% (males) of total administered radioactivity was excreted in the urine and 37.7 and 33.1% in feces. HPLC analysis of urine from both sexes revealed that 0.03-0.59% of administered [3H]EGCG, along with at least five metabolites, was excreted. In addition, we found that a second, equal administration to female mice after a 6 h interval enhanced tissue levels of radioactivity in blood, brain, liver, pancreas, bladder and bone 4-6 times above those after a single administration. These results suggest that frequent consumption of green tea enables the body to maintain a high level of tea polyphenols and this paper is the first pharmacological evidence of a wide distribution of [3H]EGCG in mouse organs, indicating a similar wide range of target organs for cancer prevention in humans.

Animals↗

PubMed1998

BAZF, a novel Bcl6 homolog, functions as a transcriptional repressor.

The BCL6 gene, which has been identified from the chromosomal translocation breakpoint in B-cell lymphomas, functions as a sequence-specific transcriptional repressor. We cloned a novel Bcl6-homologous gene, BAZF (encoding Bcl6-associated zinc finger protein). The predicted amino acid sequence of BAZF indicated that the BTB/POZ domain and the five repeats of the Krüppel-like zinc finger motif are located in the NH2-terminal region and the COOH-terminal region, respectively. BAZF associated with Bcl6 at the BTB/POZ domain and localized in the nucleus. Since zinc finger motifs of BAZF were 94% identical to those of Bcl6 at the amino acid level, BAZF bound specifically to the DNA-binding sequence of Bcl6 and functioned as a transcriptional repressor. The repressor activity was associated with both the BTB/POZ domain and the middle portion of BAZF. The 17-amino-acid sequence in the middle portion was completely conserved between BAZF and Bcl6, and the conserved region was critical for the repressor activity. Expression of BAZF mRNA, like that of Bcl6 mRNA, was induced in activated lymphocytes as an immediate-early gene. Therefore, the biochemical character of BAZF is similar to that of Bcl6 although the tissue expression pattern of BAZF differs from that of Bcl6. This is apparently the first report of a gene family whose members encode zinc finger proteins with the BTB/POZ domain.

Amino Acid Sequence↗

PubMed1998

Localization of cyclooxygenase-2 and regulation of its mRNA expression in gastric ulcers in rats.

It has been reported that cyclooxygenase-2 (COX-2) may play a crucial role in gastric ulcer healing. We examined the localization of COX-2 and the regulation of COX-2 mRNA expression in acetic acid ulcers in rats. PGE2 production was elevated in ulcerated tissue but not in intact tissue. COX-2 mRNA expression was induced in only the ulcerated tissue, and COX-2 protein was found in fibroblasts, monocytes/macrophages, and granulocytes. A selective COX-2 inhibitor inhibited increased PGE2 production by the ulcerated tissue. Interleukin-1beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha), and transforming growth factor-beta1 (TGF-beta1) mRNAs were also expressed only in the ulcerated tissue. In a culture of isolated ulcer base, blockade of IL-1beta and TNF-alpha reduced COX-2 mRNA expression and PGE2 production. In contrast, COX-2 mRNA expression and PGE2 production were promoted by prevention of TGF-beta1 action. These results indicate that COX-2 protein is highly localized in the base of gastric ulcers in rats and that COX-2 mRNA expression might be regulated positively by IL-1beta and TNF-alpha and negatively by TGF-beta1.

Acetic Acid↗

PubMed1998

Mechanism by which orally administered leminoprazole stimulates mucus synthesis in rats.

We examined the mechanism by which leminoprazole (an acid pump inhibitor) stimulates mucus synthesis in rats. Leminoprazole and omeprazole were administered to rats, and then the gastric fragments were isolated from the fundus of their stomachs. Mucus synthesis was assessed by [3H]leucine and [3H]glucosamine incorporation by the fragments into high-molecular-weight materials. After oral administration of leminoprazole, mucus synthesis by the gastric fragments was dose-dependently stimulated, while acid secretion was inhibited. Omeprazole had no effect on mucus synthesis, although it potently inhibited acid secretion. Treatment with NG-nitro-L-arginine methyl ester (L-NAME), but not indomethacin, caused a reduction of leminoprazole-stimulated synthesis of gastric mucus in a dose-dependent manner. The inhibitory action of L-NAME was restored by the concomitant administration of L-arginine, but not D-arginine. Intragastric administration of leminoprazole to pylorus-ligated rats also stimulated mucus synthesis without inhibiting acid secretion. In contrast, in the case of intraperitoneal administration, leminoprazole exerted a potent antisecretory effect, but failed to promote mucus synthesis. Exposure of leminoprazole to the luminal surface of the gastric mucosa did not cause any damage. We conclude that orally administered leminoprazole, via direct effect toward the gastric mucosa, stimulates mucus synthesis in rats probably through nitric oxide mediation. The stimulatory effect of leminoprazole on mucus synthesis is independent of its antisecretory effect.

16,16-Dimethylprostaglandin E2↗