Shizuoka Cancer Center Research Institute

Shizuoka-shi, Japan

Shizuoka Cancer Center Research Institute

Shizuoka-shi, Japan
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Ohshima K.,Shizuoka Cancer Center Research Institute | Inoue K.,Shizuoka Cancer Center Research Institute | Fujiwara A.,Shizuoka Cancer Center Research Institute | Hatakeyama K.,Shizuoka Cancer Center Research Institute | And 8 more authors.
PLoS ONE | Year: 2010

Background: Exosomes play a major role in cell-to-cell communication, targeting cells to transfer exosomal molecules including proteins,mRNAs, and microRNAs (miRNAs) by an endocytosis-like pathway.miRNAs are small noncoding RNA molecules on average 22 nucleotides in length that regulate numerous biological processes including cancer pathogenesis and mediate gene downregulation by targetingmRNAs to induce RNA degradation and/or interferingwith translation. Recent reports imply that miRNAs can be stably detected in circulating plasma and serum since miRNAs are packaged by exosomes to be protected from RNA degradation. Thus, profiling exosomal miRNAs are in need to clarify intercellular signaling and discover a novel disease marker as well. Methodology/Principal Findings: Exosomes were isolated from cultured cancer cell lines and their quality was validated by analyses of transmission electron microscopy and western blotting. One of the cell lines tested, a metastatic gastric cancer cell line, AZ-P7a, showed the highest RNA yield in the released exosomes and distinctive shape in morphology. In addition, RNAs were isolated from cells and culture media, and profiles of these three miRNA fractions were obtained using microarray analysis. By comparing signal intensities of microarray data and the following validation using RT-PCR analysis, we found that let-7 miRNA family was abundant in both the intracellular and extracellular fractions from AZ-P7a cells, while low metastatic AZ-521, the parental cell line of AZ-P7a, as well as other cancer cell lines showed no such propensity. Conclusions/Significance: The enrichment of let-7 miRNA family in the extracellular fractions, particularly, in the exosomes from AZ-P7a cells may reflect their oncogenic characteristics including tumorigenesis and metastasis. Since let-7 miRNAs generally play a tumor-suppressive role as targeting oncogenes such as RAS and HMGA2, our results suggest that AZ-P7a cells release let-7 miRNAs via exosomes into the extracellular environment to maintain their oncogenesis. © 2010 Ohshima et al.


Hatakeyama K.,Shizuoka Cancer Center Research Institute | Ohshima K.,Shizuoka Cancer Center Research Institute | Fukuda Y.,Tokyo University of Agriculture and Technology | Ogura S.-I.,Tokyo Institute of Technology | And 3 more authors.
Proteomics | Year: 2011

Splicing variation enhances proteome diversity and modulates cancer-associated proteins. Thus, the identification of alternative splice forms is significant for discovery of new cancer-related biomarkers. However, relatively few screening approaches of alternative splicing via proteomics have been reported. In the present study, we describe a combined analysis with proteome and transcriptome to simultaneously identify cancer-related splicing variants and splicing variant-derived protein fragments that are differentially expressed in a highly metastatic gastric cancer cell line MKN45P versus its parental cell line MKN45. We found three potential alternative-spliced genes using MS-based shotgun method and two different microarray platforms. Among them, aldolase C, fructose-bisphosphate (ALDOC) was predicted to have novel alternative splice forms. We successfully identified and validated novel splice forms of ALDOC gene by RT-PCR and DNA sequencing analyses, the expression level of which were higher in MKN45P than in MKN45. Furthermore, the protein fragment derived from the validated splicing variant was identified using custom-built data set including sequences of ALDOC variants in MS/MS analysis. Our combined analysis will be a promising technique for screening of cancer-related splicing variants and their protein isoforms. © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.


Nagaoka T.,Shizuoka Cancer Center Research Institute | Nagaoka T.,Waseda University | Nakamura A.,Waseda University | Okutani H.,Waseda University | And 2 more authors.
Skin Research and Technology | Year: 2012

Background: Early detection and proper excision of the primary lesions of malignant melanoma (MM) are crucial for reducing melanoma-related deaths. To support the early detection of melanoma, automated melanoma screening systems have been extensively studied and developed. In this article, we present a hyperspectral melanoma screening system and propose a possible melanoma discrimination index derived from the characteristics of the pigment molecules in the skin, both of which have been derived from hyperspectral data (HSD). Methods: The index expresses the disordered nature of each lesion including variegation in color based on variation in spectral information obtained from each lesion. Performance of the index in discriminating melanomas from other pigmented skin lesions has been studied in five cases of melanoma (41 HSD sets), one case of Spitz nevus (13 HSD sets), 10 cases of seborrheic keratosis (78 HSD sets), three cases of basal cell carcinoma (16 HSD sets), and nine cases of melanocytic nevus (21 HSD sets), obtained from patients and volunteers, all of whom were Japanese. Results: Performance of the index, which reflects the disordered nature of a lesion, discriminates melanomas with a sensitivity of 90%, a specificity of 84%, and an area under the receiver operating characteristic curve of 0.93, on resubstitution. Conclusion: An objective melanoma discrimination index at a molecular pigmentary level, derived from HSD, has been proposed, and its performance evaluated. This index was highly successful in discriminating MM from non-melanoma, although the statistical population was small. © 2011 John Wiley & Sons A/S.


Serizawa M.,Shizuoka Cancer Center Research Institute | Takahashi T.,Shizuoka Cancer Center Hospital | Yamamoto N.,Shizuoka Cancer Center Hospital | Yamamoto N.,Wakayama Medical University | Koh Y.,Shizuoka Cancer Center Research Institute
Anticancer Research | Year: 2013

Background/Aim: Mechanisms of resistance to epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitors (TKIs) in non-small cell lung cancer (NSCLC) are not fully-understood. In this study we aimed to elucidate remaining unknown mechanisms using erlotinib-resistant NSCLC cells. Materials and Methods: We performed array comparative genomic hybridization (aCGH) to identify genomic aberrations associated with EGFR-TKI resistance in erlotinib-resistant PC-9ER cells. Real-time polymerase chain reaction (PCR) and immunoblot analyses were performed to confirm the results of aCGH. Results: Among the five regions with copy number gain detected in PC-9ER cells, we focused on 22q11.2-q12.1 including v-crk avian sarcoma virus CT10 oncogene homolog-like (CRKL), the overexpression of which seemed to be associated with EGFR-TKI resistance. Blockade of downstream phosphatidylinositol 3-kinase (PI3K)/v-akt murine thymoma viral oncogene homolog (AKT) signaling using NVP-BEZ235 suppressed the proliferation of PC-9ER cells, implying the involvement of acquired CRKL amplification in EGFR-TKI resistance. Conclusion: Acquired CRKL amplification was identified as contributing to EGFR-TKI resistance; this cell line model can be utilized to study this resistance mechanism.


Yamamoto N.,Shizuoka Cancer Center | Muraakmi H.,Shizuoka Cancer Center | Nishina T.,National Shikoku Cancer Center | Hirashima T.,Osaka Prefectural Medical Center for Respiratory and Allergic Diseases | And 8 more authors.
Annals of Oncology | Year: 2013

Background: Tivantinib (formerly ARQ 197) is a selective inhibitor of c-Met mainly metabolized by CYP2C19. CYP2C19 is known for genetic polymorphisms, and ~20% of Asians are poor metabolizers (PMs), while others are extensive metabolizers (EMs). In this study, we examined the safety, pharmacokinetics (PK), and preliminary efficacy of tivantinib as a single agent to determine recommended phase II doses (RPIIDs). Patients and methods: Forty-seven patients (EMs, 33; PMs, 14) with solid tumors were orally treated with tivantinib, from 70 to 360 mg bid in a 3 + 3 dose-escalation scheme. EMs and PMs were separately enrolled at the doses >120 mg bid. Results: Tivantinib was well tolerated up to 360 mg bid for EMs and 240 mg bid for PMs. Neutropenia, leukopenia, anemia, fatigue, and anorexia were the frequent adverse events related to tivantinib and were commonly observed in both EMs and PMs. PMs had 1.9-fold higher AUC0-12 compared with EMs at 240 mg bid. Regardless of CYP2C19 phenotype, Gr.4 neutropenia occurred in patients with relatively high exposure to tivantinib. A confirmed partial response was achieved in two non-small-cell lung cancer (NSCLC) patients. Conclusion: Two different settings of RPIIDs, 360 mg bid for EMs and 240 mg bid for PMs, were determined. © The Author 2013. Published by Oxford University Press on behalf of the European Society for Medical Oncology. All rights reserved.


Kaira K.,Shizuoka Cancer Center | Serizawa M.,Shizuoka Cancer Center Research Institute | Koh Y.,Shizuoka Cancer Center Research Institute | Takahashi T.,Shizuoka Cancer Center | And 7 more authors.
Lung Cancer | Year: 2014

Background: The aim of this study is to investigate the underlying biologic mechanisms of 2-[18F]-fluoro-2-deoxy-d-glucose (18F-FDG) uptake on positron emission tomography (PET) in non-small cell lung cancer (NSCLC). Methods: One-hundred forty patients with NSCLC who underwent 18F-FDG PET were included in the study. Tumor sections were stained by immunohistochemistry for glucose transporter 1 (GLUT1), GLUT3, hypoxia-inducible factor-1 alpha (HIF-1α), hexokinase I, vascular endothelial growth factor (VEGF), microvessels (CD34), epidermal growth factor receptor (EGFR), and molecules relevant to PI3K/Akt/mTOR signaling pathway (PTEN, p-Akt, p-mTOR and p-S6). We also conducted in vitro studies of 18F-FDG uptake and mTOR inhibition in NSCLC cells. Results: High 18F-FDG uptake was significantly associated with poor prognosis in NSCLC patients. 18F-FDG uptake was significantly correlated with GLUT1, hexokinase I, HIF-1α, VEGF, CD34, p-Akt, p-mTOR and EGFR. PTEN expression showed inverse correlation with 18F-FDG uptake. In in vitro study, 18F-FDG uptake was markedly decreased by the inhibition of GLUT1 and GLUT1 upregulation by the induction of HIF-1α increased the 18F-FDG uptake. Inhibition of both mTOR complex1 (mTORC1) and mTORC2 suppressed cell growth, but activity of mTORC1 regulated the 18F-FDG uptake. NCI-H1650 cells with PTEN loss showed the highest 18F-FDG uptake and the least sensitivity to mTOR inhibitors. Conclusion: The amount of 18F-FDG accumulation is associated with molecules relevant to glucose metabolism, hypoxia, angiogenesis and mTOR signaling pathway. Especially, PTEN status may affect not only 18F-FDG uptake but also effect of mTOR inhibitors on the growth of NSCLC. © 2013 Elsevier Ireland Ltd.


Urakami K.,Shizuoka Cancer Center Research Institute | Zangiacomi V.,Shizuoka Cancer Center Research Institute | Yamaguchi K.,Shizuoka Cancer Center Research Institute | Kusuhara M.,Shizuoka Cancer Center Research Institute
Biomedical Research | Year: 2010

We used capillary electrophoresis time-of-flight mass spectrometry (CE-TOFMS) to characterize the metabolic profiles of the seed, pulp, stem and leaf of Illicium anisatum. CE-TOFMS detected more than 1000 polar metabolites within 40 min, of which 58 were annotated and quantified. Seed had higher levels of glycolytic metabolites than pulp, stem and leaf, while leaf had higher levels of TCA cycle and nucleoside metabolites. Among amino acid metabolites, levels of Gin, Glu, and Asp were higher in almost every organ. Levels of shikimic acid, the source compound for Tamiflu®, were high in all organs, ranging from 6.84% to 28.82%. These results indicate that CE-TOFMS-based metabolomics offers an efficient, convenient method for comprehensive metabolite profiling, and may be a powerful tool for the screening of drug discovery.


Maruyama K.,Shizuoka Cancer Center Research Institute | Selmani Z.,Shizuoka Cancer Center Research Institute | Ishii H.,Shizuoka Cancer Center Research Institute | Yamaguchi K.,Shizuoka Cancer Center Research Institute
International Immunopharmacology | Year: 2011

Classical cancer immunotherapy utilizes the immune response against microbial components, and a sequence of immune responses produce antitumor effects. The identification of mammalian Toll-like receptors (TLRs), receptors for microbial components, has shed light on antigen recognition by the innate immune system and provided a molecular basis for our understanding of the relationship between innate immunity and antitumor activity. However, accumulating evidence has revealed another important role of TLRs in maintaining tissue homeostasis and has also shown that tumor cells utilize this function to create favorable conditions for growth and survival, suggesting that TLR signaling acts as a double-edged sword in cancer therapy. In this review, innate immunity-based cancer therapy will be discussed with special reference to TLR-targeting drugs. © 2010 Elsevier B.V.


Kitamura Y.,Shizuoka Cancer Center Research Institute
Japan Journal of Nursing Science | Year: 2010

Aim: In order to support patients' decision-making regarding cancer treatments, it is important to clarify which criteria that cancer patients use to set priorities in their treatment choices. Using the analytic hierarchy process (AHP), a mathematical decision-making method, this article investigates the criteria and the priorities of patients with gynecological cancer. Methods: In the AHP, multiple and hierarchical criteria in the decision-making process were organized by a repeated pairwise judgment of the participants so as to serialize the alternatives along with the rational order of the priorities. For the alternatives "to receive treatment" and "to not receive treatment," the following five criteria were set: "anxiety about relapse and metastasis", "distress about side-effects", "advice of family", "advice of medical staff", and "economic burden". The participants determined a pairwise priority scale, as well as a priority scale between the alternatives for every criterion. The logical consistency of their answers was checked by a consistency index (CI). The participants were 31 patients with ovarian or endometrial cancer who were being followed up after undergoing surgery and adjuvant chemotherapy. Results: Of the participants who answered the questionnaire, 17 satisfied the logical consistency. Of the five criteria for the treatment choices, "anxiety about relapse and metastasis" and "advice of medical staff" were found to be the important factors for treatment choice; however, the weight attached to the priority criteria differed much among the patients. Conclusion: The AHP made it possible to support patients' decision-making in order to clarify their priority criteria and to quantitatively present their decision-making process. © 2010 The Author. Journal compilation © 2010 Japan Academy of Nursing Science.


Vincent Z.,Shizuoka Cancer Center Research Institute | Urakami K.,Shizuoka Cancer Center Research Institute | Maruyama K.,Shizuoka Cancer Center Research Institute | Yamaguchi K.,Shizuoka Cancer Center Research Institute | Kusuhara M.,Shizuoka Cancer Center Research Institute
Genes and Cancer | Year: 2014

During the past decade, cancer stem-like cells (CSCs) have drawn substantial interest in cancer research since they have been described as major targets to improve treatment of tumors and to prevent recurrence and metastasis. In this paper, we report on the search for CSCs within the Colo205 human adenocarcinoma cell line. We describe that CD133 (prominin) was the only reliable marker for the isolation and characterization of CSCs within a Colo205 cell population. CD133-positive cells displayed many CSC characteristics, such as tumorsphere formation ability, expression of early-stage development markers, high invasiveness, raised tumor initiation potential and resistance to cisplatin chemotherapy treatment. In vitro analyses also highlighted a specific metabolomic profile of CD133-positive cells and we concluded that the chemotherapy resistance of CSCs could be related to the quiescence of such cells associated with their reduced metabolism. Furthermore, in vivo metabolome analyses suggested that a high level of circulating glutathione molecules could also promote treatment resistance. From the perspective of metabolomics, we also discuss the controversial use of serum-free in vitro cultures for CSC enrichment prior to further phenotype characterization. © 2014, Impact Journals LLC. All rights reserved.

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