研究者詳細 - 豊國　伸哉

論文 - 豊國　伸哉

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CD146 and insulin-like growth factor 2 mRNA-binding protein 3 predict prognosis of asbestos-induced rat mesothelioma. 査読有り国際誌

Okazaki Y, Nagai H, Chew SH, Li J, Funahashi S, Tsujimura T, Toyokuni S

Cancer science 104 巻 ( 8 ) 頁： 989 - 95 2013年8月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Cancer Science

Malignant mesothelioma (MM), which is associated with asbestos exposure, is one of the most deadly tumors in humans. Early MM is concealed in the serosal cavities and lacks specific clinical symptoms. For better treatment, early detection and prognostic markers are necessary. Recently, CD146 and insulin-like growth factor 2 mRNA-binding protein 3 (IMP3) were reported as possible positive markers of MM to distinguish from reactive mesothelia in humans. However, their application on MM of different species and its impact on survival remain to be elucidated. To disclose the utility of these molecules as early detection and prognostic markers of MM, we injected chrysotile or crocidolite intraperitoneally to rats, thus obtaining 26 peritoneal MM and establishing 11 cell lines. We immunostained CD146 and IMP3 using paraffin-embedded tissues and cell blocks and found CD146 and IMP3 expression in 58% (15/26) and 65% (17/26) of MM, respectively, but not in reactive mesothelia. There was no significant difference in both immunostainings for overexpression among the three histological subtypes of MM and the expression of CD146 and IMP3 was proportionally associated. Furthermore, the overexpression of CD146 and/or IMP3 was proportionally correlated with shortened survival. These results suggest that CD146 and IMP3 are useful diagnostic and prognostic markers of MM. © 2013 Japanese Cancer Association.

DOI： 10.1111/cas.12185

Lewis y antigen is expressed in oral squamous cell carcinoma cell lines and tissues, but disappears in the invasive regions leading to the enhanced malignant properties irrespective of sialyl-Lewis x. 査読有り国際誌

Hotta H, Hamamura K, Yamashita K, Shibuya H, Tokuda N, Hashimoto N, Furukawa K, Yamamoto N, Hattori H, Toyokuni S, Ueda M, Furukawa K

Glycoconjugate journal 30 巻 ( 6 ) 頁： 585 - 97 2013年8月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Glycoconjugate Journal

Expression and implication of carbohydrate antigens in squamous cell carcinomas (SCCs) in oral cavity was examined. In the cell lines, type 2H and Lewis y antigens were markedly expressed. In the tissues from SCC patients and benign disorders, type 2H was highly expressed in hyperplasia (96.4 %), displasia (92.9 %) and SCC (100 %). Lewis y was, in turn, expressed mainly in cancer tissues (91.3 %), suggesting that Lewis y is a cancer-associated antigen. Normal oral mucosa showed no expression of these blood group antigens. Surprisingly, Lewis y antigen disappeared in the invasion sites where Ki-67 was definitely stained. Over-expression of Lewis y with manipulation of a fucosyltransferase cDNA resulted in suppression of cell growth and invasion, and knockdown of Lewis y also brought about increased cell growth and invasion. In either situations, no changes in the expression of sialyl-Lewis x could be found. Lowered tumor growth and invasion into surrounding tissues were also shown in Lewis y-positive SCC grafts in nu/nu mice. All these results together with alternative staining between Lewis y and Ki-67 in cancer tissues and FUT1 transfectants suggested that loss of Lewis y is a crucial event for the late stage of SCCs. © 2012 Springer Science+Business Media New York.

DOI： 10.1007/s10719-012-9458-2

Evaluation of two distinct methods to quantify the uptake of crocidolite fibers by mesothelial cells. 査読有り国際誌

Yamashita K, Nagai H, Kondo Y, Misawa N, Toyokuni S

Journal of clinical biochemistry and nutrition 53 巻 ( 1 ) 頁： 27 - 35 2013年7月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：一般社団法人　日本酸化ストレス学会

Exposure to asbestos fibers increases the risk of mesothelioma in humans. One hypothetical carcinogenic mechanism is that asbestos fibers may directly induce mutations in mesothelial cells. Although the uptake of asbestos fibers by mesothelial cells is recognized, methods for the quantification of the uptake have not been well established. In the present study, we evaluated two distinct methods, using crocidolite fibers and MeT5A mesothelial cells. One method is histological evaluation using the cell-block technique, which allows for the direct cross-sectional observation of cells and fibers. We found the bright field observation with ×1000 magnification (oil-immersion) of the sample with Kemechtrot staining was most suitable for this purpose. The other method is flow cytometric analysis, which permits the evaluation of a much larger number of cells. We observed that the side scatter (SSC) increased with the intracellular fibers, and that the "mean SSC ratio (treated/control)" was useful for quantification. We could collect the cells with abundant internalized crocidolite fibers by sorting. Results of the two methodologies were correlated well in the experiments. The quantities of internalized fibers increased with incubation time and loaded dosage, but they were inversely associated with cellular density in culture. © 2013 JCBN.

DOI： 10.3164/jcbn.12-104

Repeated short-term daily exercise ameliorates oxidative cerebral damage and the resultant motor dysfunction after transient ischemia in rats. 査読有り

Hamakawa M, Ishida A, Tamakoshi K, Shimada H, Nakashima H, Noguchi T, Toyokuni S, Ishida K

Journal of clinical biochemistry and nutrition 53 巻 ( 1 ) 頁： 8 - 14 2013年7月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：一般社団法人　日本酸化ストレス学会

Long-term exercise prior to brain ischemia enhances the activities of antioxidant enzymes and leads to a significant reduction in brain damage and neurological deficits in rats subjected to transient middle cerebral artery occlusion. However, it has not been established whether relatively short-term exercise generates similar results following middle cerebral artery occlusion. We aimed to determine whether short-term exercise could reduce oxidative damage and prevent sensori-motor dysfunction. Male Wistar rats were subjected to perform daily exercise on a treadmill for 30 min at a speed of 15 m/min for 3 weeks, followed by a 90-min middle cerebral artery occlusion. Animals were assessed after middle cerebral artery occlusion for neurological deficits and sensori-motor function. Brain tissues were processed to evaluate infarct volume and oxidative damage. Oxidative stress was assessed using immunohistochemistry for 4-hydroxy-2-nonenalmodified proteins and 8-hydroxy-2'-deoxyguanosine. Antioxidant enzymes were evaluated using immunohistochemistry for thioredoxin and activity assay for superoxide dismutase. Exercise for 3 weeks decreased the severity of paralysis and impairment in forelimb motor coordination. Furthermore, exercise had effect on superoxide dismutase and reduced the infarct volume and the number of cells immunopositive for 4-hydroxy-2-nonenal-modified proteins and 8-hydroxy-2'-deoxyguanosine. Our results suggest that pre-conditioning treadmill exercise for 3 weeks is useful for ameliorating ischemia-induced brain injury. © 2013 JCBN.

DOI： 10.3164/jcbn.12-72

Multispecificity of immunoglobulin M antibodies raised against advanced glycation end products: involvement of electronegative potential of antigens. 査読有り国際誌

Chikazawa M, Otaki N, Shibata T, Miyashita H, Kawai Y, Maruyama S, Toyokuni S, Kitaura Y, Matsuda T, Uchida K

The Journal of biological chemistry 288 巻 ( 19 ) 頁： 13204 - 14 2013年5月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Journal of Biological Chemistry

Background: Advanced glycation end products (AGEs) can act as neoantigens to trigger immune responses. Results: Natural IgM antibodies against AGEs recognize multiple molecules, including DNA and chemically modified proteins. Conclusion: There is a close relationship between the formation of AGEs and innate immune responses. Significance: Our findings highlight AGEs and related modified proteins as a source of multispecific natural antibodies. Copyright © 2013 by The American Society for Biochemistry and Molecular Biology, Inc.

DOI： 10.1074/jbc.M113.452177

DOI： 10.1074/jbc.m113.452177

CANCER AS A FERROTOXIC DISEASE: WHAT WE HAVE LEARNED FROM ANIMAL STUDIES TOWARD ITS PREVENTION 国際誌

Toyokuni, S; Jiang, L; Okazaki, Y; Akatsuka, S

AMERICAN JOURNAL OF HEMATOLOGY 88 巻 ( 5 ) 頁： E168 - E169 2013年5月

Met is the most frequently amplified gene in endometriosis-associated ovarian clear cell adenocarcinoma and correlates with worsened prognosis. 査読有り国際誌

Yamashita Y, Akatsuka S, Shinjo K, Yatabe Y, Kobayashi H, Seko H, Kajiyama H, Kikkawa F, Takahashi T, Toyokuni S

PloS one 8 巻 ( 3 ) 頁： e57724 2013年3月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：PLoS ONE

Clear cell adenocarcinoma of the ovary (OCC) is a chemo-resistant tumor with a relatively poor prognosis and is frequently associated with endometriosis. Although it is assumed that oxidative stress plays some role in the malignant transformation of this tumor, the characteristic molecular events leading to carcinogenesis remain unknown. In this study, an array-based comparative genomic hybridization (CGH) analysis revealed Met gene amplification in 4/13 OCC primary tumors and 2/8 OCC cell lines. Amplification of the AKT2 gene, which is a downstream component of the Met/PI3K signaling pathway, was also observed in 5/21 samples by array-based CGH analysis. In one patient, both the Met and AKT2 genes were amplified. These findings were confirmed using fluorescence in situ hybridization, real-time quantitative PCR, immunoblotting, and immunohistochemistry. In total, 73 OCC cases were evaluated using real-time quantitative PCR; 37.0% demonstrated Met gene amplification (>4 copies), and 8.2% had AKT2 amplification. Furthermore, stage 1 and 2 patients with Met gene amplification had significantly worse survival than patients without Met gene amplification (p<0.05). Met knockdown by shRNA resulted in reduced viability of OCC cells with Met amplification due to increased apoptosis and cellular senescence, suggesting that the Met signaling pathway plays an important role in OCC carcinogenesis. Thus, we believe that targeted inhibition of the Met pathway may be a promising treatment for OCC. © 2013 Yamashita et al.

DOI： 10.1371/journal.pone.0057724

FROM NAGOYA TO THE WORLD 国際誌

Toyokuni, S

NAGOYA JOURNAL OF MEDICAL SCIENCE 75 巻 ( 1-2 ) 頁： 1 - 2 2013年2月

Protective effect of cardioplegia with poly (ADP-ribose) polymerase-1 inhibitor against myocardial ischemia-reperfusion injury: in vitro study of isolated rat heart model. 査読有り国際誌

Yamazaki K, Tanaka S, Sakata R, Miwa S, Oriyanhan W, Takaba K, Minakata K, Marui A, Ikeda T, Toyokuni S, Komeda M, Ueda K

Journal of enzyme inhibition and medicinal chemistry 28 巻 ( 1 ) 頁： 143 - 7 2013年2月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Journal of Enzyme Inhibition and Medicinal Chemistry

Poly(ADP-ribose) polymerase (PARP)-1 inhibitor has been suggested to attenuate the ischemia-reperfusion injury. We investigated the protective effect of the cardioplegia with a PARP-1 inhibitor, 4-hydoxyquinazoline (4-HQ), against myocardial ischemia-reperfusion injury. Isolated rat hearts were perfused on a Langendorff apparatus and cardioplegically arrested for 90 min by perfusion with St. Thomas' Hospital solution (ST-solution). In the Group ST (n = 8), the hearts were arrested with the ST-solution alone. The Group HQ (n = 8) were treated with the ST-solution containing 4-HQ (10 μM) for cardioplegia. During reperfusion, the Group HQ showed significantly greater functional recovery of +dp/dtmax (p = 0.005) and lower enzymatic leakage (p < 0.01). NAD+ levels were also preserved higher in the Group HQ (p < 0.01). Immunohistochemical study revealed lesser extents of oxidative stress and apoptosis, in the Group HQ. Thus, addition of 4-HQ in the cardioplegia may provide a new intervention for myocardial protection against ischemia-reperfusion injury by decreasing NAD+ consumption and suppressing oxidative stress. © 2013 Informa UK, Ltd.

DOI： 10.3109/14756366.2011.642373

FROM NAGOYA TO THE WORLD 国際誌

Toyokuni, S

NAGOYA JOURNAL OF MEDICAL SCIENCE 75 巻 ( 1-2 ) 頁： 1 - 2 2013年2月

Deferasirox induces mesenchymal-epithelial transition in crocidolite-induced mesothelila carcinogenesis in rats. 査読有り

Nagai H, Okazaki Y, Chew SH, Misawa N, Yasui H and Toyokuni S.

Cancer Prev Res 6 巻 ( 11 ) 頁： 1222-1230 2013年

Genotoxicity and carcinogenicity risk of carbon nanotubes. 査読有り

Toyokuni S.

Adv Drug Deliver Rev 2013年

Repeated short-term daily exercise ameliorates oxidative cerebral damage and the resultant motor dysfunction after transient ischemia in rats. 査読有り

Hamakawa M, Ishida A, Tamakoshi K, Shimada H, Nakashima H, Noguchi T, Toyokuni S and Ishida K.

J Clin Biochem Nutr 53 巻頁： 8-14 2013年

CD146 and IMP3 predict prognosis of asbestos-induced rat mesothelioma. 査読有り

Okazaki Y, Nagai H, Chew SH, Jiang L, Funahashi S, Tsujimura T and Toyokuni S.

Cnacer Sci 104 巻頁： 989-995 2013年

Multi-specificity of IgM antibodies raised against advanced glycation end products: Involvement of electronegative potential of antigens. 査読有り

Chikazawa M, Otaki N, Shibata T, Miyashita H, Kawai Y, Maruyama S, Toyokuni S, Kitaura Y, Matsuda T, Uchida K.

J Biol Chem 288 巻頁： 13204-13214 2013年

Met is the most frequently amplified gene in endometriosis-associated ovarian clear cell adenocarcinoma and correlates with worsened prognosis. 査読有り

Yamashita Y, Akatsuka S, Shinjo K, Yatabe Y, Kobayashi H, Seko H, Kajiyama H, Kikkawa F, Takahashi T and Toyokuni S.

PLoS One 8 巻 ( 3 ) 頁： e57724 2013年

Evaluation of two distinct methods to quantify the uptake of crocidolite fibers by mesothelial cells. 査読有り

Yamashita K, Nagai H, Kondo Y, Misawa N and Toyokuni S.

J Clin Biochem Nutr 53 巻頁： 27-35 2013年

Metamorphosis of mesothelial cells with active horizontal motility in tissue culture. 査読有り

Nagai H, Chew SH, Okazaki Y, Funahashi S, Namba T, Kato T, Enomoto A, Jiang L, Akatsuka S and Toyokuni S.

Sci Rep 3 巻頁： 1144 2013年

Protective effect of cardioplegia with poly(ADP-ribose)polymerase-1 inhibitor aganst myocardial ischemia-reperfusion injury: in vitro study if isolated rat heart model. 査読有り

Yamazaki K, Tanaka S, Sakata R, Miwa S, Oriyanhan W, Takaba K, Minakata K, Marui A, Ikeda T, Toyokuni S, Komeda M and Ueda K.

J Enzyme Inhibition Medicinal Chem 28 巻頁： 143-147 2013年

Hotta H, Hamamura K, Yamashita K, Shibuya H, Tokuda N, Hashimoto N, Furukawa K, Yamamoto N, Hattori H, Toyokuni S, Ueda M, Furukawa K.

Glycoconj J 30 巻頁： 585-597 2013年

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