Toshio Kokuryo

Nagoya University, Nagoya, Aichi, Japan

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Publications (47)163.43 Total impact

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    ABSTRACT: Special AT-rich sequence-binding protein 1 and 2 (SATB1/2) are nuclear matrix-associated proteins involved in chromatin remodeling and regulation of gene expression. SATB2 acts as a tumor suppressor in laryngeal squamous cell carcinoma and colon cancer, whereas SATB1 promotes the progression of numerous types of cancers. In this study, we examined the effects of SATB1 and SATB2 on the malignant characteristics of colorectal cancer cells. SATB1 and SATB2 expression were negatively correlated in colorectal cancer specimens. SATB1 expression was increased, whereas SATB2 expression was reduced, in colorectal cancer tissues compared to control tissues. Exogenous expression of SATB2 in colorectal cancer cells suppressed cell proliferation, colony formation and tumor proliferation in mice. c-Myc was reduced by SATB2 expression, and exogenous expression of c-Myc in SATB2-expressing cells restored proliferation, colony formation and in vivo tumor growth of colorectal cancer cells. We also showed that c-Myc reduction by SATB2 was mediated by the inactivation of ERK5. In contrast, SATB1 promoted c-Myc expression. The expression of SATB1 in colorectal cancer tissues was positively correlated with c-Myc expression, and SATB1 knockdown reduced c-Myc expression in colorectal cancer cells. Finally, we showed that SATB1 knockdown in colorectal cancer cells suppressed cell proliferation, colony formation and cell invasion. Our results reveal interesting features of how the structural homologs SATB1 and SATB2 exert opposing functions in colorectal tumorigenesis.
    Full-text · Article · Dec 2015 · Oncotarget
  • T Takagi · Y Yokoyama · T Kokuryo · J Yamaguchi · M Nagino
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    ABSTRACT: Background Surgical treatment for perihilar cholangiocarcinoma frequently involves hepatectomy and extrahepatic bile duct resection with a choledochojejunostomy (CJ). Cholangitis owing to bilioenteric anastomosis is a common complication. The impact of CJ or regurgitating cholangitis on the liver regeneration process after major hepatectomy is unknown. Methods Rats underwent 70 per cent hepatectomy (Hx group) or hepatectomy with CJ (Hx + CJ group). The intrahepatic inflammatory response, hepatic regeneration rate, and expression of regeneration-associated genes in the liver and blood were compared between these two groups. ResultsLevels of hepatobiliary markers in the blood were significantly higher 4 and 7 days after operation in the Hx + CJ group than the Hx group. Intrahepatic expression of inflammation-associated genes, such as interleukin 6 and tumour necrosis factor , was also significantly higher in the Hx + CJ group on days 4 and 7. A progressive periportal inflammatory response was identified in the Hx + CJ group by histological examination. The hepatic regeneration rate was significantly lower in the Hx + CJ group than in the Hx group on day 2 (mean(s.d.) 142(63) versus 214(26) per cent; P = 0013) and day 4 (324(53) versus 413(44) per cent; P = 0004). Gene expression levels of hepatic regeneration-promoting factors such as hepatocyte growth factor were significantly lower in the Hx + CJ group than the Hx group on day 1. Conclusion: CJ perturbs early liver regeneration after hepatectomy. An excessive inflammatory response in the liver and suppression of liver regeneration-associated factors may play a role.
    No preview · Article · Aug 2015 · British Journal of Surgery

  • No preview · Article · Aug 2015 · Cancer Research
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    ABSTRACT: The targeting protein for Xklp2 (TPX2) is a microtubule- and, cell cycle-associated protein who's overexpression has been reported in various malignancies. In this study, we verified the overexpression of TPX2 in both surgically resected specimens of pancreatic cancer and multiple pancreatic cancer cell lines. Subsequently, we found that TPX2 siRNA effectively suppressed the proliferation of pancreatic cancer cells in culture, and the direct injection of TPX2 siRNA into subcutaneously implanted pancreatic cancer cells in nude mice revealed antiproliferative effects. These results implied a therapeutic potential of TPX2 siRNA in pancreatic cancer. Among 56 angiogenesis-related factors examined using angiogenesis arrays, the average protein levels of insulin-like growth factor-binding protein-3 (IGFBP-3) were significantly higher in TPX2 siRNA-treated tumors than in the Control siRNA-treated tumors. Moreover, we demonstrated that CD34-positive microvessels were significantly reduced in tumors treated with TPX2 siRNA compared to tumors that treated with Control siRNA. The attenuated expression of CD34 in TPX2 siRNA-treated tumors coincided with the overexpression of IGFBP-3. These results indicated that TPX2 has an impact on tumor angiogenesis in pancreatic cancer. The results also implied that the antiangiogenic effect observed in TPX2 siRNA-treated pancreatic cancer cells may be partly explained by the upregulation of IGFBP-3. © 2015 The Authors. Cancer Medicine published by John Wiley & Sons Ltd.
    Preview · Article · Apr 2015 · Cancer Medicine

  • No preview · Conference Paper · Apr 2015
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    ABSTRACT: Special AT-rich sequence binding protein 2 (SATB2) is an evolutionarily conserved transcription factor that has multiple roles in neuronal development, osteoblast differentiation, and craniofacial patterning. SATB2 binds to the nuclear matrix attachment region (MAR) and regulates the expression of diverse sets of genes by altering chromatin structure. Recent studies have reported that the high expression of SATB2 is associated with favorable prognosis in colorectal and laryngeal cancer; however, it remains uncertain whether SATB2 has tumor-suppressive functions in cancer cells. In this study, we examined the effects of SATB2 expression on the malignant characteristics of colorectal cancer cells. The expression of SATB2 repressed the proliferation of cancer cells in vitro and in vivo and also suppressed their migration and invasion. Extracellular signal regulated kinase 5 (ERK5) is a MAP kinase that is associated with an aggressive phenotype in various types of cancer. SATB2 expression reduced the activity of ERK5, and constitutive activation of ERK5 restored the proliferation, anchorage-independent growth, migration, and invasion of SATB2-expressing cells. Our results show a novel regulatory mechanism of SATB2-mediated tumor suppression via ERK5 inactivation. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.
    Full-text · Article · Feb 2015 · FEBS Journal
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    ABSTRACT: Objectives This randomized clinical trial was designed to investigate whether inchinkoto has a hepatoprotective effect on postoperative outcome after major hepatectomy.Methods Sixty-one patients scheduled for major hepatectomy were randomly assigned to one of two groups in which preoperative inchinkoto was (inchinkoto group, n = 30) or was not (non-inchinkoto group, n = 31) administered. Inchinkoto was administered for at least 7 days before surgery. The primary endpoint was the incidence of post-hepatectomy liver damage. The expression of nuclear factor E2-related factor 2 (Nrf2) and other oxygen stress-related markers in the liver were also determined.ResultsThere was no significant difference in clinical characteristics between the inchinkoto and non-inchinkoto groups. Serum levels in liver function tests and incidences of post-hepatectomy liver failure did not differ significantly between the two groups. However, there was a significantly higher induction of antioxidant factors in the liver, such as Nrf2 protein and heme oxygenase-1 mRNA, after hepatectomy in the inchinkoto group than in the non-inchinkoto group.Conclusions The preoperative administration of inchinkoto did not have a significant impact on the overall outcome of major hepatectomy. However, inchinkoto induced the expression of Nrf2 during hepatectomy and may have exerted an antioxidative effect on the liver.
    No preview · Article · Feb 2015 · HPB
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    ABSTRACT: RNA splicing is a fundamental process for protein synthesis. Recent studies have reported that drugs that inhibit splicing have cytotoxic effects on various tumor cell lines. In this report, we demonstrate that depletion of SNW1, a component of the spliceosome, induces apoptosis in breast cancer cells. Proteomics and biochemical analyses revealed that SNW1 directly associates with other spliceosome components, including EFTUD2 (Snu114) and SNRNP200 (Brr2). The SKIP region of SNW1 interacted with the N-terminus of EFTUD2 as well as two independent regions in the C-terminus of SNRNP200. Similar to SNW1 depletion, knockdown of EFTUD2 increased the numbers of apoptotic cells. Furthermore, we demonstrate that exogenous expression of either the SKIP region of SNW1 or the N-terminus region of EFTUD2 significantly promoted cellular apoptosis. Our results suggest that the inhibition of SNW1 or its associating proteins may be a novel therapeutic strategy for cancer treatment. © 2014 The Authors. Cancer Medicine published by John Wiley & Sons Ltd.
    Full-text · Article · Feb 2015 · Cancer Medicine
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    ABSTRACT: Although an aggressive surgical approach to perihilar cholangiocarcinoma (PHC) has improved survival, a prognosis of advanced PHC remains unsatisfactory. The overexpression of mesenchymal-epithelial transition factor (MET) and recepteur d'origine nantais (RON) has been shown to be associated with poor prognosis in some types of cancer. One hundred sixty-nine patients who underwent histologically curative resection for PHC were subjected to immunohistochemical analysis for MET and RON. The association between a positive expression of MET or RON and clinicopathologic features as well as the patients' prognosis were analyzed. There were 27 patients (16 %) who had a positive expression for both MET and RON. Although clinicopathologic features in the either MET- or RON-negative group were not significantly different compared to the both MET- and RON-positive group, the prognosis tended to be worse in the patients with both MET and RON positivity. When the analysis was limited to patients with advanced-stage disease (stage III and IVa), a multivariate analysis revealed that both MET and RON positivity and lymph node metastasis were identified as independent poor prognostic factors. The overall survival rate for patients with both MET and RON positivity was worse than that with either MET or RON negativity in patients with advanced PHC. The poor prognosis in these patients was not associated with unfavorable clinicopathologic features. The examination of MET and RON expression in PHC may enable a tailored method for patient classification that could not otherwise be achieved using the conventional pathologic classification system.
    No preview · Article · Jan 2015 · Annals of Surgical Oncology

  • No preview · Article · Oct 2014 · Gan to kagaku ryoho. Cancer & chemotherapy
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    ABSTRACT: In summary, we showed that SATB2 has a tumor-suppressive function in colorectal cancer cells. The exogenous expression of SATB2 suppressed the aggressive phenotype of colorectal cancer cells. In addition, SATB2 induced the inactivation of ERK5, and the constitutive activation of ERK5 restored the malignant phenotype of SATB2-expressing cells. MAP kinase phosphatases like DUSP10 and PPM1A-2 were up regulated in SATB2 expressing cells. This effect may be responsible for the inactivation of MEK5/ERK5 pathway. Cell lines: Colorectal cancer (CRC) cell lines; HT29, HCT116 and DLD-1 were obtained from ATCC and cultured in the recommended media. Stable cell lines: were generated by retroviral infection using the pVPack-GP and pVPack-Ampho vectors with Lipofectamine 2000 (Invitrogen, Carlsbad, CA). Xenograft tumor assay: 6 nude mice (six weeks) were subcutaneously injected with 1x10 6 DLD-1 cells and subsequent tumor volume and final tumor weight were determined. Colony formation assay: 1x10 4 CRC cells in different groups were mixed with agar/DMEM and 10%FBS for 2 weeks and then colony number and size were determined. Immunoblotting: GFP, SATB2, p-ERK5, ERK5, c-Myc, FLAG and β-actin were identified by immunoblotting. Quantitative PCR analysis: RNA were extracted and cDNA were generated and then mRNAs of different genes were quantified using Real Time System TP800 (TAKARA). Cell proliferation assay: viable cells were counted in 96 well plate using cell count kit 8.
    Full-text · Conference Paper · Sep 2014
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    ABSTRACT: Hilar cholangiocarcinoma is clinically characterized by biliary obstruction in the porta hepatis. Because the boundary between the intrahepatic and extrahepatic bile duct is unclear, hilar cholangiocarcinoma can potentially arise from both ducts. Therefore, the definition of hilar cholangiocarcinoma remains under debate. In November 2013, the 6th edition of the General Rules for Clinical and Pathological Studies on Cancer of the Biliary Tract was released, following the American Joint Committee on Cancer (AJCC) or International Union Against Cancer (UICC) TNM system. In that edition, as an alternative to "hilar cholangiocarcinoma," the new term "perihilar cholangiocarcinoma" is defined as cholangiocarcinoma involving the perihilar bile duct, despite the presence or absence of a significant liver mass component. This definition clearly indicates that some intrahepatic as well as extrahepatic perihilar tumors are involved in the perihilar tumor category. From the clinical point of view, there is no need for a differential diagnosis between intrahepatic or extrahepatic tumors therefore, the new definition is readily applicable in multidisciplinary team management. Japanese clinicians were previously required to distinguish between the proper use of the AJCC/UICC and the Japanese staging systems, but now the current revision will allow the more convenient use of a single, globally standardized staging system in daily practice.
    No preview · Article · Jul 2014 · Nippon Geka Gakkai zasshi
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    ABSTRACT: The objective of this study was to elucidate the role of toll-like receptor 4 (TLR4) in liver injury induced by biliary obstruction and subsequent intraportal lipopolysaccharide (LPS) infusion in rats. Biliary obstruction often leads to the development of bacterial translocation. Rats were subjected to either a sham operation (Sham group) or bile duct ligation for 7 days (BDL group). Seven days after each operation, LPS (0.5 μg) was injected through the ileocecal vein. In other experiments, rats that had undergone BDL were pretreated, prior to LPS challenge, with internal biliary drainage (Drainage group); intravenous TAK-242, a TLR4 inhibitor (TAK group); or intravenous GdCl3, a Kupffer cell deactivator (GdCl3 group). The expression of the TLR4 protein as well as the number of Kupffer cells in the liver were significantly increased in the BDL group compared with the Sham group. These changes were normalized after biliary drainage. The expression of TLR4 co-localized with Kupffer cells, which was confirmed by double immunostaining. Serum levels of liver enzymes and proinflammatory cytokines after intraportal LPS injection were significantly higher in the BDL group than in the Sham group. However, pretreatment with TAK-242 or GdCl3 strongly attenuated these changes to levels similar to those seen with biliary drainage.These results imply that blocking TLR4 signaling effectively attenuates liver damage to the same level as that observed with biliary drainage in rats with BDL and subsequent intraportal LPS infusion. TAK-242 treatment may be used for patients who are susceptible to liver damage by biliary obstruction and endotoxemia.
    Preview · Article · Dec 2013 · AJP Gastrointestinal and Liver Physiology
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    ABSTRACT: In patients with luminal-type breast cancer (positive for ER and/or PgR), a complete consensus on the threshold indication for a combination of chemotherapy and endocrine therapy has not been achieved, especially for patients with HER2-negative luminal type (HNLT). Girdin, an actin-binding Akt substrate, plays a crucial role in the migration of cancer cells. This study examined the expression of Girdin in relation to clinicopathological features and other immunohistochemical markers (HER2, Ki-67), especially in patients with HNLT breast cancer. One hundred one breast cancer patients who underwent surgery were evaluated. Immunohistochemical staining was performed for Girdin and other biomarkers, such as ER, PgR, HER2, and Ki-67. Positive expression of Girdin was observed in 26 patients. The expression of Girdin was significantly associated with the incidence of lymph node metastases (p = 0.001). Among the other examined biomarkers, positive expression of Ki-67 also showed a significant association with the incidence of lymph node metastases (p = 0.04). In the HNLT breast cancer patients (n = 73), the 5-year recurrence-free survival rate was significantly lower (57 %) in patients with positive expression of both Girdin and Ki-67 than the rate in other patients (92 %) (p = 0.002). This study demonstrated that the expression of Girdin in invasive breast cancer is strongly associated with lymph node metastasis. The expression status of Girdin and Ki-67 can be a useful biomarker in stratifying patients with HNLT breast cancer into those with high risk of recurrence and the need for additional chemotherapy.
    No preview · Article · Oct 2013 · Breast Cancer
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    ABSTRACT: The International Study Group of Liver Surgery (ISGLS) has defined bile leakage as a drain fluid-to-serum total bilirubin concentration (TBC) ratio (the bilirubin ratio) ≥3.0. The aim of the present study was to determine the clinical significance of this definition, and to outline characteristics of bile leakage in complex hepatectomy. The TBCs of the serum and drain fluid were measured on postoperative days (POD) 1, 3, and 7 in 241 patients who had undergone hepatobiliary resection. The validation of the bilirubin ratio and predictors of bile leakage were retrospectively assessed. Grade A, B, or C bile leakage was found in 23 (9.5 %), 66 (27.4 %), and 0 patients, respectively. The median duration of drainage was 27 days in grade B bile leakage. The sensitivity and specificity of the bilirubin ratio for detecting grade B bile leakage were 59 and 87 %, respectively. The area under the receiver operating characteristics curve of the drain fluid TBC on POD 3 had the highest predictive value: 68 % sensitivity and 76 % specificity for a drain fluid TBC of 3.7 mg/dL. The multivariate analysis demonstrated that operative time, left trisectionectomy, bilirubin ratio, and TBC of the drain fluid on POD 3 were independent predictors of grade B bile leakage. In complex hepatectomy, bile leakage develops most frequently after left trisectionectomy and often results in a refractory clinical course. The ISGLS biochemical definition is valid, and a combination of bilirubin ratio and drain fluid TBC may enhance risk prediction for grade B bile leakage.
    No preview · Article · Oct 2013 · World Journal of Surgery
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    ABSTRACT: Inchinkoto (ICKT) is one of the most commonly used herbal medicines and is a hepatoprotective agent. Among the numerous chemical compounds included in ICKT, geniposide is the most abundant component. After oral intake, geniposide is converted into the active metabolite genipin by intestinal bacteria and absorbed in the portal circulation. The biological properties of ICKT and its major active ingredient genipin have been studied in numerous experiments using cells and animals. ICKT or genipin administration exerts a choleretic effect through upregulation of multidrug resistance-associated protein 2 in hepatocytes. ICKT also exerts antiapoptoic activity by inhibiting the transforming growth factor beta 1- or tumor necrosis factor alpha-dependent signaling pathway. The excessive inflammatory response induced by various forms of hepatic stress is also attenuated by ICKT preadministration. Proinflammatory cytokine-induced upregulation of inducible nitric oxide synthase is strongly attenuated by ICKT in both in vivo and in vitro experiments. Moreover, ICKT enhances antioxidant enzymes in the liver under oxidative stress. These experimental results clearly indicate the effects of ICKT on hepatic stress. To date, however, clinical data on the benefits of ICKT for liver disease are very rare. To extend the clinical applications of ICKT in humans, it is crucial to design and perform a rigorous clinical trial. In this review article, recent evidence relating to the hepatoprotective effects of ICKT in the field of basic and clinical science is summarized and discussed.
    No preview · Article · Sep 2013 · Nippon Geka Gakkai zasshi
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    ABSTRACT: Background: The aim of this study was to determine the intrahepatic kinetics of different types of nitric oxide (NO) synthase, such as endothelial nitric oxide synthase (eNOS) and inducible NOS (iNOS), during repeated ischemia/reperfusion (I/R). Methods: Three different protocols of hepatic I/R in rats were designed as follows: 60 min of ischemia and 30 min of reperfusion (I/R 60/30); 5 min of ischemia and 5 min of reperfusion (I/R 5/5); and repeating 15 min of ischemia and 5 min of reperfusion for four cycles (I/R 15/5 × 4). Intrahepatic NO levels were measured using a selective NO sensor. Changes in hepatic microcirculation during I/R 5/5 were investigated using intravital microscopy. Hepatic expression of eNOS, phospho-eNOS, and iNOS were evaluated during repeated I/R by Western blot, reverse transcription polymerase chain reaction, and immunohistochemistry. Results: During I/R 60/30, intrahepatic NO levels gradually increased and then reached a plateau approximately 15 min after starting ischemia. During I/R 5/5, the sinusoids after 5 min reperfusion were dilated compared with the sinusoids before ischemia. The expression of phospho-eNOS during I/R 15/5 × 4 markedly increased during the first ischemia, and then the levels attenuated during the subsequent repeating I/R cycles; however, the expression of iNOS gradually increased, as observed by Western blot, reverse transcription polymerase chain reaction, and immunohistochemical analysis. An impact of NO production by phospho-eNOS activation during the superacute phase of I/R was also confirmed using pharmacologic NOS inhibitors. Conclusion: Our results firstly demonstrated an altered activation of the phospho-eNOS system and iNOS over the course of repeated hepatic I/R.
    No preview · Article · Feb 2013 · Journal of Surgical Research
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    ABSTRACT: Objective: We determined whether there is a protective effect of branched-chain amino acid (BCAA) on hepatic ischemia/reperfusion (IR)-induced acute liver injury. Methods: Wister rats were divided into four groups: simple laparotomy with vehicle; simple laparotomy with BCAA (1 g/kg of body weight orally); IR (30 min clamp) with vehicle; and IR with BCAA. Serum liver function tests and the gene expression of adhesion molecules (ICAM and VCAM) and vasoconstrictor-related genes (endothelin-1) in the liver were examined. In the in vivo study, portal venous pressure, leukocyte adhesion, and hepatic microcirculation were evaluated. Furthermore, Kupffer cells were isolated and cultured with various concentrations of BCAA in the presence or absence of lipopolysaccharide (LPS). Results: Increased levels of liver function tests following IR were significantly attenuated by BCAA treatment. The increased expression of adhesion molecules and endothelin-1 were also significantly attenuated by BCAA treatment. Moreover, increased portal venous pressure, enhanced leukocyte adhesion, and deteriorated hepatic microcirculation following IR were all improved by BCAA treatment. In the experiment using isolated Kupffer cells, the expression of interleukin-6, interleukin-1β, and endothelin-1 in response to LPS stimulation was attenuated by BCAA in a dose-dependent fashion. Conclusions: These results indicate that perioperative oral administration of BCAA has excellent therapeutic potential to reduce IR-induced liver injury. These beneficial effects may result from the direct attenuation of Kupffer cell activation under stressful conditions.
    No preview · Article · Dec 2012 · AJP Gastrointestinal and Liver Physiology
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    ABSTRACT: Objective: To investigate the expression of calcitonin gene-related peptide (CGRP) and its role in the liver regeneration process after 70% hepatectomy (Hx). Materials and methods: Wistar rats were divided into eight groups based on time after Hx. Remnant liver samples were collected serially 0 h, 1 h, 6 h, 12 h, 1 d, 2 d, 7 d, and 14 d after Hx (n = 6 for each time point). The expression level of the calcitonin/CGRP gene in the remnant liver was measured. Western bolts and immunohistochemistry were performed to determine the levels of CGRP in the regenerating liver. Furthermore, CGRP8-37 (a CGRP receptor antagonist) was used to examine the role of CGRP during liver regeneration. Results: A marked upregulation of the calcitonin/CGRP gene was observed immediately after Hx, and the protein levels of CGRP in the liver, which were measured by western blot and immunohistochemistry, also rapidly increased after Hx. The liver regeneration rate was significantly attenuated by an administration of CGRP8-37 2 d after Hx. The mitotic index was evaluated by histologic examination 1 and 2 d after Hx and was also significantly lower in the CGRP8-37 group. In addition, CGRP8-37 treatment inhibited the phosphorylation of extracellular-signal regulated kinase 1/2. The levels of early response genes, such as c-fos, c-jun, and c-myc, were also downregulated by CGRP8-37. Conclusion: The calcitonin/CGRP gene may have an important role in the early phase of liver regeneration after Hx.
    No preview · Article · Dec 2012 · Journal of Surgical Research

  • No preview · Article · Oct 2012 · Gan to kagaku ryoho. Cancer & chemotherapy