Institute for Advanced Medical science

Nishinomiya, Japan

Institute for Advanced Medical science

Nishinomiya, Japan

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Nagaya H.,Institute for Advanced Medical science | Gotoh A.,Institute for Advanced Medical science | Kanno T.,Hyogo College of Medicine | Tsuchiya A.,Hyogo College of Medicine | Nishizaki T.,Hyogo College of Medicine
Pharmacology | Year: 2014

Naftopidil, an 1-adrenoceptor blocker, induced apoptosis of human malignant pleural mesothelioma NCI-H2052 cells. Naftopidil upregulated the expression of tumor necrosis factor- (TNF-) mRNA in these cells. Naftopidil, alternatively, increased FasL secretion from NCI-H2052 cells, without affecting the expression of FasL mRNA and protein, and activated caspase-3 and -8 in NCI-H2052 cells. Naftopidil drastically suppressed tumor growth in mice inoculated with these cells. The results of the present study indicate that naftopidil induces apoptosis of NCI-H2052 cells by upregulating the expression of TNF- and stimulating the secretion of FasL, a ligand for the death receptor Fas, both to activate caspase-8 and the effector caspase-3, leading to the suppression of NCI-H2052 cell proliferation in vivo. This raises the possibility that naftopidil could be developed as an effective drug for the treatment of malignant pleural mesothelioma. © 2014 S. Karger AG, Basel.

Eguchi R.,Institute for Advanced Medical science | Eguchi R.,Hyogo College of Medicine | Kubo S.,Hyogo College of Medicine | Takeda H.,Institute for Advanced Medical science | And 5 more authors.
Carcinogenesis | Year: 2012

Malignant mesothelioma is an aggressive tumor arising from mesothelial cells of serous membranes. Src family kinases (SFKs) have a pivotal role in cell adhesion, proliferation, survival and apoptosis. Here, we examined the effect of SFK inhibitors in NCI-H2052, ACC-MESO-4 and NCI-H28 cells, mesothelioma cell lines and Met5A, a human non-malignant mesothelial cell line. We found that PP2, a selective SFK inhibitor, inhibited SFK activity and induced apoptosis mediated by caspase-8 in NCI-H28 but not Met5A, NCI-H2052 and ACC-MESO-4 cells. Src, Yes, Fyn and Lyn protein, which are members of the SFK, were expressed in these cell lines, whereas NCI-H28 cells were deficient in Fyn protein. Small interfering RNA (siRNA) targeting Fyn facilitated PP2-induced apoptosis mediated by caspase-8 in NCI-H2052 and ACC-MESO-4 cells. PP2 reduced Lyn protein levels and suppressed SFK activity in all mesothelioma cell lines. Lyn siRNA induced caspase-8 activation and apoptosis in NCI-H28 cells but not in NCI-H2052 and ACC-MESO-4 cells. However, double RNA interference knockdown of Fyn and Lyn induced apoptosis accompanied by caspase-8 activation in NCI-H2052 and ACC-MESO-4 cells. Dasatinib, an inhibitor of multi-tyrosine kinases including SFK, also inhibited SFK activity and induced reduction of Lyn protein levels, caspase-8 activation and apoptosis in NCI-H28 cells but not in other cell lines. Present study suggests that SFK inhibitors induce caspase-8-dependent apoptosis caused by reduction of Lyn protein in Fyn-deficient mesothelioma cells. © The Author 2012. Published by Oxford University Press. All rights reserved.

Yamamoto H.,Institute for Advanced Medical science | Okuzaki D.,Chip Development Center for Infectious Diseases | Yamanishi K.,Hyogo College of Medicine | Xu Y.,Institute for Advanced Medical science | And 13 more authors.
International Journal of Molecular Medicine | Year: 2013

Spontaneously hypertensive rats (SHR) and stroke-prone SHR (SHRSP) are frequently used as model rats not only in studies of essential hypertension and stroke, but also in studies of attention deficit hyperactivity disorder (ADHD). Normotensive Wistar-Kyoto rats (WKY) are normally used as controls in these studies. In this study, using these rats, we aimed to identify the genes causing hypertension and stroke, as well as the genes involved in ADHD. Since adrenal gland products can directly influence cardiovascular, endocrine and sympathetic nervous system functions, gene expression profiles in the adrenal glands of the 3 rat strains were examined using genome-wide microarray technology when the rats were 3 and 6 weeks of age, a period in which the rats are considered to be in a pre-hypertensive state. Gene expression profiles were compared between SHR and WKY and between SHRSP and SHR. A total of 353 genes showing more than a 4-fold increase or less than a 4-fold decrease in expression were isolated and candidate genes were selected as significantly enriched genes. SHR-specific genes isolated when the rats were 3 weeks of age contained 12 enriched genes related to transcriptional regulatory activity and those isolated when the rats were 6 weeks of age contained 6 enriched genes related to the regulation of blood pressure. SHRSP-specific genes isolated when the rats were 3 weeks of age contained 4 enriched genes related to the regulation of blood pressure and those isolated when the rats were 6 weeks of age contained 4 enriched genes related to the response to steroid hormone stimulus. Ingenuity pathway analysis of enriched SHR-specific genes revealed that 2 transcriptional regulators, cAMP responsive element modulator (Crem) and Fos-like antigen 1 (Fosl1), interact with blood pressure-regulating genes, such as neurotensin (Nts), apelin (Apln) and epoxide hydrolase 2, cytoplasmic (Ephx2). Similar analyses of SHRSP-specific genes revealed that angiotensinogen (Agt), one of the blood pressure-regulating genes, plays pivotal roles among SHRSP-specific genes. Moreover, genes associated with ADHD, such as low density lipoprotein receptor (Ldlr) and Crem, are discussed.

Li W.,Institute for Advanced Medical science | Li W.,Hyogo College of Medicine | Kubo S.,Institute for Advanced Medical science | Okuda A.,Institute for Advanced Medical science | And 7 more authors.
Journal of Immunotherapy | Year: 2010

Zoledronate (Zol) has recently been shown to expand γδ T cells that play important roles in host defenses against infection and tumors. In this study, we examined effects of interleukin-18 (IL-18) on expansion of γδ T cells in human peripheral blood mononuclear cells (PBMCs) stimulated by Zol and IL-2. The expansion of γδ T cells stimulated by Zol and IL-2 was strongly promoted by exogenous IL-18, and to the contrary, inhibited by neutralizing anti-IL-18 receptor antibody. The γδ T cells that expanded in the presence of Zol, IL-2, and IL-18 exhibited the phenotype of effector memory cells characterized by CD44 (+), CD27 (-), and CD45RA (-). In addition, they expressed NKG2D, perforin, CD94, CD25, and CD122, and 15% to 40% of them were positive for CD56. Incubation of γδ T cells in the presence with IL-18 produced GM-CSF, IFN-γ, and TNF-α at much higher levels than those incubated without IL-18. They showed strong cytotoxicity against tumor cells including mesothelioma cells and inhibited growth of xenograft of mesothelioma in mice. These observations indicate that IL-18 can efficiently promote expansion of γδ T cells with potent antitumor activity. Copyright © 2010 by Lippincott Williams & Wilkins.

PubMed | Institute for Advanced Medical science
Type: Journal Article | Journal: Journal of immunology (Baltimore, Md. : 1950) | Year: 2011

T cells are considered to be innate lymphocytes that play an important role in host defense against tumors and infections. We recently reported that IL-18 markedly amplified T cell responses to zoledronate (ZOL)/IL-2. In an extension of this finding, we analyzed the mechanism underlying the IL-18-mediated expansion of T cells. After incubation of PBMCs with ZOL/IL-2/IL-18, the majority of the cells expressed TCR, and the rest mostly exhibited CD56(bright)CD11c(+) under the conditions used in this study. CD56(bright)CD11c(+) cells were derived from a culture of CD56(int)CD11c(+) cells and CD14(+) cells in the presence of IL-2 and IL-18 without the addition of ZOL. They expressed IL-18Rs, HLA-DR, CD25, CD80, CD83, CD86, and CD11a/CD18. In addition, they produced IFN-, TNF-, but not IL-12, when treated with IL-2/IL-18, and they exerted cytotoxicity against K562 cells, thus exhibiting characteristics of both NK cells and dendritic cells. Incubation of purified T cells with CD56(bright)CD11c(+) cells in the presence of ZOL/IL-2/IL-18 resulted in the formation of massive cell clusters and led to the marked expansion of T cells. However, both conventional CD56(-/int)CD11c(high) dendritic cells induced by GM-CSF/IL-4 and CD56(+)CD11c(-) NK cells failed to support the expansion of T cells. These results strongly suggest that CD56(bright)CD11c(+) cells play a key role in the IL-18-mediated proliferation of T cells.

PubMed | Hyogo College of Medicine, Institute for Advanced Medical science and Institute of Biomedical Research and Innovation
Type: Journal Article | Journal: Stem cells (Dayton, Ohio) | Year: 2015

Brain vascular pericytes (PCs) are a key component of the blood-brain barrier (BBB)/neurovascular unit, along with neural and endothelial cells. Besides their crucial role in maintaining the BBB, increasing evidence shows that PCs have multipotential stem cell activity. However, their multipotency has not been considered in the pathological brain, such as after an ischemic stroke. Here, we examined whether brain vascular PCs following ischemia (iPCs) have multipotential stem cell activity and differentiate into neural and vascular lineage cells to reconstruct the BBB/neurovascular unit. Using PCs extracted from ischemic regions (iPCs) from mouse brains and human brain PCs cultured under oxygen/glucose deprivation, we show that PCs developed stemness presumably through reprogramming. The iPCs revealed a complex phenotype of angioblasts, in addition to their original mesenchymal properties, and multidifferentiated into cells from both a neural and vascular lineage. These data indicate that under ischemic/hypoxic conditions, PCs can acquire multipotential stem cell activity and can differentiate into major components of the BBB/neurovascular unit. Thus, these findings support the novel concept that iPCs can contribute to both neurogenesis and vasculogenesis at the site of brain injuries.

PubMed | Institute for Advanced Medical science
Type: Journal Article | Journal: Experimental and therapeutic medicine | Year: 2012

CUB-domain-containing protein 1 (CDCP1)/CD318 is a single transmembrane molecule highly expressed in colorectal cancer and leukemia. It has also been shown to be expressed in hematopoietic progenitor cells. In this study, we analyzed the expression of CD318 on cord blood hematopoietic stem and progenitor cells. Cord blood mononuclear cells were depleted of mature blood cell linage (Lin)-positive cells and then Lin-negative cells were sorted by flow cytometry based on the expression of CD34 and CD318. Analysis of sorted cells by colony-forming assay showed that CD34(+)CD318(+) cells produced more mixed colony forming units and erythroid burst forming unit-derived colonies than CD34(+)CD318(-) cells. These colonies were also produced by CD34(-)CD318(+) and CD34(-)CD318(-) cells, but were generally fewer in number. When sorted cells were cultured on a monolayer of human mesenchymal stem cells, CD34(+)CD318(+) cells proliferated more abundantly than CD34(+)CD318(-) cells, while CD34(-)CD318(+) and CD34(-)CD318(-) cells failed to proliferate. Transplantation of CD34(+)CD318(+) cells into non-obese diabetic/severe combined immunodeficient disease (NOD/SCID) mice resulted in efficient reconstitution of human cells, indicating that CD34(+)CD318(+) cells possess strong SCID-repopulating cell activity. These findings suggest that the co-expression of CD34 and CD318 identifies the immature character of hematopoietic stem cells.

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