PubMed Health⌕ Search

Biomedical subjects

Hiromitsu Saisho

Publications and source records attributed to Hiromitsu Saisho.

At least 73 records · Page 4Linked to original sources
PubMed2005

Methylation status of p14ARF, p15INK4b, and p16INK4a genes in human hepatocellular carcinoma.

BACKGROUND: The INK4 locus consisting of three genes involved in the regulation of cell cycle, p16INK4a, p15INK4b, and p14ARF is often disrupted in human neoplasms. METHODS: We analyzed the promoter methylation of each gene by methylation-specific PCR in hepatocellular carcinoma (HCC). RESULTS: The methylation of p16INK4a, p15INK4b, and p14ARF was found to occur in 27 (69.2%), seven (17.9%), and none out of 39 HCC tumors, respectively. Regarding corresponding nontumorous liver tissues, the promoter regions of p16INK4a, p15INK4b, and p14ARF were methylated in three (17.6%), three (17.6%), and none out of 17 samples, respectively. Analysis of mRNA expression revealed that loss of p16INK4a expression was frequently observed in HCC. In contrast, transcripts of p14ARF and p15INK4b were detected in 16 (88.9%) and 16 (88.9%) of 18 tumors, respectively. CONCLUSIONS: The frequent loss of transcription of p16INK4a with promoter methylation not only in the advanced but also in the early stages of HCC suggests that the epigenetic alteration of p16INK4a promoter is likely to be involved in hepatocarcinogenesis. Together with the result of RT-PCR analysis, the role of aberrant methylation of p14ARF or p15INK4a promoter in hepatocarcinogenesis is thought to be limited.

Adult↗
PubMed2005

Evaluation of clinical usefulness of second-generation HCV core antigen assay: comparison with COBAS AMPLICOR HCV MONITOR assay version 2.0.

BACKGROUND: Hepatitis C virus (HCV) is an important etiologic agent for chronic liver diseases. METHODS: The aim of this study was to evaluate the clinical usefulness of second-generation HCV core antigen assay by comparing the results of the assay with those of the COBAS AMPLICOR HCV MONITOR version 2.0 (COBAS v2.0). RESULTS: HCV core antigen was detectable by this assay in 142/149 (95.3%) of serotype 1 (3821+/-322 fmol/l; mean+/-SD), in 56/58 (96.6%) of serotype 2 (2589+/-449 fmol/l), and in 6/6 (100%) of serotypes 1+2 (1240+/-548 fmol/l). The HCV core antigen levels measured by this assay correlated well with the HCV RNA levels by COBAS v2.0 (r=0.848, P<0.0001). In relation to the outcome of interferon monotherapy, the pretreatment HCV core antigen levels of sustained and non-sustained virological responders were 659+/-189 and 4904+/-376 fmol/l in serotype 1, 1993+/-740 and 3145+/-519 fmol/l in serotype 2. The cutoff values with the best accuracy for HCV core Ag levels to discriminate between sustained and non-sustained virological response were 699 fmol/l for serotype 1 and 292 fmol/l for serotype 2, respectively, by receiver operating characteristic curve analysis. CONCLUSION: This new assay was considered to be useful in evaluating the HCV levels in patients with chronic hepatitis C.

Adolescent↗
PubMed2005

Fas polymorphisms influence susceptibility to autoimmune hepatitis.

BACKGROUND AND AIMS: Genetic factors associated with autoimmune hepatitis (AIH) and primary biliary cirrhosis (PBC), immune-mediated chronic inflammatory liver diseases of unknown etiology, remain to be elucidated. Polymorphisms of the gene encoding Fas have been linked to a variety of autoimmune diseases. We hypothesized that Fas gene polymorphisms might be genetic markers for AIH and PBC. METHODS: To determine the frequency and significance of Fas polymorphisms in patients with AIH and PBC, 74 Japanese AIH patients, 98 Japanese PBC patients, and 132 ethnically matched control subjects were investigated by the use of the Taqman assay. RESULTS: We found significant differences between AIH patients and controls in allele frequencies of Fas-670 (p=0.009), Fas IVS (intervening sequence) 2nt176 (p=0.018), Fas IVS3nt46 (p=0.031), and Fas IVS5nt82 (p=0.013) polymorphisms. Haplotype analysis revealed that one of the haplotypes, GATGC, was associated with increased AIH prevalence. On the other hand, we found no statistically significant differences between PBC patients and controls in allele frequencies of the Fas polymorphisms genotyped in this study. CONCLUSIONS: These results indicate a genetic link of Fas polymorphisms to the development of AIH. Further studies are needed to determine the genetic factors contributing to the development of AIH.

Adolescent↗
PubMed2005

Evaluation of distribution patterns for copper and zinc in metallothionein and superoxide dismutase in chronic liver diseases and hepatocellular carcinoma using high-performance liquid chromatography (HPLC).

It has been reported that the copper (Cu) content of hepatocytes increases in chronic liver diseases and small hepatocellular carcinoma (HCC). In cells, Cu exists mainly as Cu-metallothionein (MT) or Cu, zinc (Zn)-superoxide dismutase (SOD). In this study, we investigated the biochemical state of Cu in the hepatocytes of patients with HCC using high-performance liquid chromatography (HPLC). The subjects of present study were 23 patients with HCC who underwent liver resection. The cancerous tissue and non-cancerous hepatic parenchyma with chronic disease were analyzed. In addition, as a normal control, hepatic tissue was collected at autopsy from 13 patients with no liver disease. Each sample was diluted with buffer, chilled, homogenized, and centrifuged. The supernatant was fractionated using HPLC. The metal contents of each fraction were measured using a desktop-type inductively coupled plasma (ICP) emission spectrochemical analyzer. HPLC analysis showed that MT existed mainly as Zn-MT in the normal hepatic tissue. The case of Cu,Zn-MT was significantly greater than Zn-MT in the non-cancerous, but diseased hepatic parenchyma than in the normal hepatic tissue (p<0.01). In comparison with non-cancerous hepatic parenchyma, the Cu-MT in the cancerous section was significantly greater than the Cu,Zn-MT (p<0.01). The Cu content for MT was significantly higher in small HCC (<40 mm) (p<0.01), and the absence of Cu or Zn in the MT fraction was significantly more frequent in the large HCC (>or=40 mm) (p<0.01). The Cu and Zn content for SOD in the samples showed no significant difference. Increase in the Cu content in the cancerous hepatic tissue were, thought to be reflecting changes in the distribution of Cu in the MT fraction of hepatic tissues.

Aged↗
PubMed2005

Assessment of portal vein invasion in pancreatic cancer by fusion 3-dimensional ultrasonography.

OBJECTIVE: The purpose of this study was to assess the usefulness of a newly developed imaging technique, fusion 3-dimensional ultrasonography (3DUS) in the diagnosis of portal vein (PV) invasion in patients with pancreatic cancer (PC). METHODS: Fourteen patients with proven PC were examined by fusion 3DUS presented as shaded volume-rendering and multiplanar reconstruction images. The surgical findings were obtained in all patients (12 with resection and 2 without). The findings were compared with those of 2-dimensional ultrasonography (2DUS), contrast-enhanced computed tomography (CT), dynamic CT (DCT), angiography, and surgical findings. Portal vein invasion was assessed by 3 independent radiologists for each modality, and objectivity of the assessment was examined by interobserver variability analysis (kappa value). RESULTS: On the basis of surgical findings, the accuracy rates of 2DUS, fusion 3DUS, DCT, and angiography were 78.6%, 92.9%, 85.3%, and 66.7%, respectively. The kappa values of 2DUS, fusion 3DUS, DCT, and angiography for PV invasion were 0.57, 0.90, 0.63, and 0.49, respectively, being most objective in fusion 3DUS. CONCLUSIONS: Fusion 3DUS is useful for diagnosis of PV invasion of PC.

Adult↗
PubMed2005

5-aza-2'-deoxycytidine sensitizes hepatoma and pancreatic cancer cell lines.

Hepatocellular carcinoma (HCC) and pancreatic cancer are at the forefront of chemotherapy-resistant tumors with poor prognosis. Even with innovative treatment regimens, response rates remain low and the duration of response is short. We examined whether the suppression of DNA methylation was capable of enhancing the sensitivity of hepatoma and pancreatic cancer cell lines to 5-fluorouracil (5-FU). 5-aza-2'-deoxycytidine (5-aza-dC) at 2 microM, a specific DNA methylation inhibitor, did not induce cell death in Huh-7 with or without HCV, HLE, HepG2 and MIA PaCa-2 cells. However, a combination of 5-aza-dC with 5-FU showed a reduction in cell viability and induction of apoptosis in these cell lines to a greater degree than with 5-FU only. These findings underline the fact that DNA methylation plays a key role in conferring chemoresistance to hepatoma and pancreatic cancer, and the combination of DNA methylation inhibitor with chemotherapy could be a novel and highly effective tool for future targeted therapy of chemoresistant tumors.

Antimetabolites, Antineoplastic↗
PubMed2005

[A pilot study of TS-1+CDDP therapy for highly advanced stage IV gastric cancer].

We performed a pilot study of combination chemotherapy with TS-1 and cisplatin for highly advanced gastric cancer. From June 2002, 12 patients with multiple liver metastases, carcinomatous lymphangitis or peritoneal dissemination, were enrolled in the study. TS-1 was administered at a daily dose on day 1-21 and an intermediate-dose of cisplatin (60 mg/m2) was administered on day 8. The combination was repeated in a 5-week cycle. The median administered cycles were three (one to eight). An objective response was obtained in 9 cases (75.0%) of primary sites and 6 cases of metastatic sites. No severe hematological toxicity occurred, and grade 3 stomatitis (in one case) and vomiting (in two cases) occurred as non-hematological toxicities. The improvement of clinical symptoms such as appetite loss and abdominal discomfort was obtained in 9 of 10 cases. The median survival time is 244 days. The TS-1/CDDP regimen had almost no survival benefits, but may induce relief of symptoms due to cancer and better quality of life.

Administration, Oral↗
PubMed2004

Hypermethylation of NAD(P)H: quinone oxidoreductase 1 (NQO1) gene in human hepatocellular carcinoma.

BACKGROUND/AIMS: NAD(P)H: quinone oxydoreductase 1 (NQO1) and glutathione S-transferase P1 (GSTP1) belong to phase II xenobiotic-metabolizing enzymes. GSTP1 inactivation via CpG island hypermethylation in hepatocellular carcinoma (HCC) was previously reported, but the involvement of NQO1 in HCC is not well known. In this study, we assessed the transcription and status of methylation of NQO1 gene in human hepatoma cells and primary human HCC tissues. METHODS: NQO1 transcription and DNA hypermethylation in hepatoma cells with or without 5-aza-deoxycytidine (5-Aza-CdR) treatment were investigated by reverse-transcription PCR (RT-PCR), sodium bisulfite sequencing and methylation-specific PCR (MSP). The methylation status of NQO1 and GSTP1, and NQO1 mRNA in 44 HCC cases was also analyzed by MSP and real-time PCR, respectively. RESULTS: NQO1 transcription was down-regulated and the CpG island DNA was hypermethylated in Hep3B and HuH6 cells. After treatment with 5-Aza-CdR, NQO1 transcription was restored and CpG island DNA was demethylated in these cells. MSP analysis revealed that NQO1 hypermethylation occurred in 50.0% of HCC. All of the tumors that exhibited lesser amounts of NQO1 mRNA than corresponding non-tumorous tissues showed NQO1 hypermethylation. CONCLUSIONS: NQO1 transcription might be inappropriately suppressed by promoter hypermethylation in a subset of HCC, as well as GSTP1 gene.

Acetylation↗
PubMed2004

Gene expression profiles in liver regeneration with oval cell induction.

The liver has the unique ability to regenerate even in adulthood. While mature hepatocytes can proliferate by themselves, stem cells also play a critical role in liver regeneration and oval cells are considered to be the progeny of activated hepatic stem cells. We herein investigated the gene expression profiles in the conditions inducing oval cells, using microarray analysis. Two approaches were used to induce oval cells. In the first, animals were treated with a combination of an N-2-acetylaminofluorene (AAF)-containing diet and partial hepatectomy (PHx). In the second, animals were supplied with chow containing a 1:1 mixture of choline-deficient and normal diets, as well as 0.075% ethionine in drinking water. Using in-house cDNA microarrays consisting of 2304 cDNA clones from the mouse liver, 69 and 89 genes, respectively, were found to be up-regulated in these two models. Six genes, i.e., those for insulin-like growth factor binding protein-1, CYP4a14, carnitine octanoyltransferase, osteopontin, and two expressed sequence tags (ESTs) were up-regulated in these models but not in ordinary model with PHx alone. They might be specifically activated in the induction of oval cells, and help to clarify the nature of stem cell stimulation that occurs during liver regeneration.

2-Acetylaminofluorene↗
PubMed2004

Impaired lymphocyte development and function in Clast5/Stra13/DEC1-transgenic mice.

Clast5/Stra13/DEC1 is a member of the helix-loop-helix family of transcriptional repressors. We have previously shown that Clast5 is rapidly down-regulated upon B cell activation and its overexpression inhibits cell cycle progression in B lymphoma cells. In the present study, we show that Clast5 expression is developmentally regulated during B cell differentiation, being expressed at the progenitor B cells, down-regulated at the precursor B cells, elevated in immature and mature resting B lymphocytes, and down-regulated again in germinal center B cells. To investigate the function of Clast5 in regulating lymphocyte development, we have generated transgenic mice expressing Clast5 in B- and T-lineage cells (Clast5-Tg). Clast5-Tg mice grew and bred normally but their spleen and thymus cellularity was reduced compared with control littermates. The development of B cells in the bone marrow and T cells in the thymus was impaired, with the expansion of progenitor B and T cells most strongly affected. The frequency of IL-7-responsive cells in the bone marrow of Clast5-Tg mice was reduced by >80% and their proliferative response to IL-7 was also compromised. Mature B cells from Clast5-Tg mice were hyporesponsive to antigen receptor cross-linking and exhibited mild reduction in the proliferative response to CD40 ligation or lipopolysaccharide stimulation. Moreover, the development of germinal center B cells and antibody production against a T-dependent antigen were reduced in Clast5-Tg mice. These results reveal a critical role for Clast5/Stra13/DEC1 in negatively regulating lymphocyte development and function in vivo.

Animals↗
PubMed2004

Immunohistochemical metallothionein expression in hepatocellular carcinoma: relation to tumor progression and chemoresistance to platinum agents.

BACKGROUND: Metallothionein (MT), which is known to detoxify heavy metal ions, is considered to serve as a mechanism of resistance to platinum complex compounds. In the present study, MT expression in hepatocellular carcinoma (HCC) was immunohistologically investigated to clarify its relationship to clinical background factors and responsiveness to anticancer drugs. METHODS: Specimens from 117 patients with HCC were immunohistologically studied, using a monoclonal anti-MT antibody. the percentage of MT-positive HCC (MT ratio) cells was determined, to evaluate the extent of staining with anti-MT antibody. Staining with an MT ratio of more than 50% was categorized as diffusely positive; an MT ratio of 5% to less than 50% was focally positive; and an MT ratio of less than 5% was negative. Twenty-two patients received repeated arterial infusion chemotherapy with carboplatin (CBDCA), a platinum-containing compound, and the MT expression was analyzed in relation to their chemotherapeutic response. RESULTS: The ratio of MT-positive cells in HCC decreased with the degree of histological differentiation and also decreased with higher tumor stage. In patients treated with CBDCA, the ratio of MT-positive cells in responders was significantly lower than that in non-responders. CONCLUSIONS: MT expression decreases with the degree of histological differentiation and decreases with increasing tumor stage in HCC. In addition, MT expression may lower the antitumor effect of CBDCA.

Antineoplastic Agents↗
PubMed2004

A case of severe acute hepatitis of unknown etiology treated with the Chinese herbal medicine Inchinko-to.

A prolonged severe hepatitis of unknown etiology was treated with Inchinko-to, a Chinese herbal medicine, and this case is herein described. Inchinko-to was given with ursodeoxycholic acid (UDCA) and glycyrrhizin. The improvement in the patient's liver function seemed to be accelerated after the treatment, especially after stopping the administration of kanamycin sulfate which might possibly inhibit the conversion of geniposide, one of the constituents of Inchinko-to, to an active ingredient through the suppression of the bacterial growth in intestinal flora, suggesting the usefulness of Inchinko-to for treatment of severe hepatitis.

Journal Article↗
PubMed2004

Identification of genes up-regulated by histone deacetylase inhibition with cDNA microarray and exploration of epigenetic alterations on hepatoma cells.

BACKGROUND/AIMS: Epigenetics is the key factor in the regulation of gene expression. We conducted cDNA microarray analysis to screen for genes induced by histone deacetylase (HDAC) inhibition and examined epigenetic alterations. METHODS: Microarray analysis was performed in six hepatoma cell lines and primary hepatocytes treated with trichostatin A (TSA). mRNA expression of several genes was examined by reverse transcription-polymerase chain reaction in TSA-treated cells and hepatoma samples. Acetylated histones and methylation status in 5'CpG islands was assessed by chromatin immunoprecipitation (ChIP) assay and bisulfite genomic sequencing, respectively. RESULTS: Fifty-seven genes showed greater than 2-fold change after TSA treatment in multiple cell lines. Among them, four genes including p21(WAF1) exhibited substantial induction (greater than 5-fold changes). Decreased mRNA levels of these genes in hepatoma tissues were observed in more than half of patients. ChIP assay, in general, demonstrated a good correlation between mRNA expression and histone acetylation, but only a limited correlation with the methylated DNA in the promoter region. CONCLUSIONS: We identified 57 up-regulated genes by TSA treatment in hepatoma cells and some of them appeared to be cancer-related genes in hepatomas. The alterations in acetylated histones are likely closely associated with gene expression.

Acetylation↗
PubMed2004

Metal contents of liver parenchyma after percutaneous ethanol injection or radiofrequency ablation in patients with hepatocellular carcinoma before and after trientine hydrochloride therapy.

We administered trientine hydrochloride, a drug used in the treatment of Wilson's disease, to patients with hepatocellular carcinoma after radical treatment with percutaneous ethanol injection or radiofrequency ablation, and examined its effect on the reduction of liver-tissue copper content. We enrolled 24 patients with 3 or fewer primary lesions of Child class A or B hepatocellular carcinoma with diameters of 3 cm or less who had undergone radical treatment with percutaneous ethanol injection or radiofrequency ablation. Trientine hydrochloride was orally administered in a single daily dose of 250 mg to 12 patients before a meal (at fasting, group 1) or at a total daily dosage of 750 mg, divided into 3 doses, to 12 patients (group 2). This study was a randomized between-groups comparative study of 12 weeks' duration. We used the particle-induced x-ray-emission method to determine liver-tissue mineral content. Urine copper and serum mineral levels were also measured, and transaminase levels were examined. Liver-tissue copper content decreased significantly, to 160.1 microg/g dry weight, after treatment, compared with the pretreatment level of 306.8 microg/g dry weight (P <.05). We detected no significant difference in iron or zinc content before and after treatment. The copper content was significantly reduced after treatment in both groups (P <.05). The urine copper level was significantly increased after 1 week of treatment but decreased thereafter. Serum copper levels were significantly reduced after treatment (P <.01). We detected no significant difference in transaminase level before and after treatment. Iron-deficiency anemia in 1 patient after 12 weeks' treatment was the only adverse reaction, and it was improved by the administration of an iron product. We noted no other overt adverse reactions. In patients with hepatocellular carcinoma, trientine hydrochloride therapy may significantly reduce copper content in liver tissue.

Aged↗