Hitoshi Nobumasa

Secreted CXCL1 Is a Potential Mediator and Marker of the Tumor Invasion of Bladder Cancer

Purpose: The purpose of this study was to identify proteins that are potentially involved in the tumor invasion of bladder cancer. Experimental Design: We searched for the candidate proteins by comparing the profiles of secreted proteins among the poorly invasive human bladder carcinoma cell line RT112 and the highly invasive cell line T24. The proteins isolated from cell culture supernatants were identified by shotgun proteomics. We found that CXCL1 is related to the tumor invasion of bladder cancer cells. We also evaluated whether the amount of the chemokine CXCL1 in the urine would be a potential marker for predicting the existence of invasive bladder tumors. Results: Higher amount of CXCL1 was secreted from highly invasive bladder carcinoma cell lines and this chemokine modulated the invasive ability of those cells in vitro. It was revealed that CXCL1 regulated the expression of matrix metalloproteinase-13 in vitro and higher expression of CXCL1 was associated with higher pathologic stages in bladder cancer in vivo. We also showed that urinary CXCL1 levels were significantly higher in patients with invasive bladder cancer (pT1-4) than those with noninvasive pTa tumors (P = 0.0028) and normal control (P < 0.0001). Finally, it was shown that CXCL1 was an independent factor for predicting the bladder cancer with invasive phenotype. Conclusions: Our results suggest that CXCL1 modulates the invasive abilities of bladder cancer cells and this chemokine may be a potential candidate of urinary biomarker for invasive bladder cancer and a possible therapeutic target for preventing tumor invasion.

High Tc Phase of Bi-Sr-Ca-Cu-O Superconductor

High Tc superconducting phase above 100 K in a Bi-Sr-Ca-Cu-O superconductor was found to appear in proportion to the time of the sintering just below the melting temperature. The changes of the magnetic susceptibility for this high Tc phase corresponded well to the behavior of 4.8° peak in X-ray diffraction pattern taken with Cu-Kα, which indicated that the high Tc phase contains triple Cu-O layers sandwiched by two Bi2O2 layers. The mechanism of the high Tc phase formation is proposed such that a disproportionation into triple and single Cu-O layers occurs from 80 K phase with double Cu-O layers to form the high Tc and a semiconducting phase, respectively.

Microarray-based identification of CUB-domain containing protein 1 as a potential prognostic marker in conventional renal cell carcinoma.

PurposeRenal cell carcinoma (RCC) is characterized by a variable and unpredictable clinical course. Thus, accurate prediction of the prognosis is important in clinical settings. We conducted microarray-based study to identify a novel prognostic marker in conventional RCC.Patients and methodsThe present study included the patients surgically treated at Kyoto University Hospital. Gene expression profiling of 39 samples was carried out to select candidate prognostic markers. Quantitative real-time PCR of 65 samples confirmed the microarray experiment results. Finally, we evaluated the significance of potential markers at their protein expression level by immunohistochemically analyzing 230 conventional RCC patients.ResultsUsing expression profiling analysis, we identified 14 candidate genes whose expression levels predicted unfavorable disease-specific survival. Next, we examined the expression levels of nine candidate genes by quantitative real-time PCR and selected CUB-domain containing protein 1 (CDCP1) for further immunohistochemical analysis. Positive staining for CDCP1 inversely correlated with disease-specific and recurrence-free survivals. In multivariate analysis including clinical/pathological factors, CDCP1 staining was a significant predictor of disease-specific and recurrence-free survivals.ConclusionsWe identified CDCP1 as a potential prognostic marker for conventional RCC. Further studies might be required to confirm the prognostic value of CDCP1 and to understand its function in RCC progression.

MicroRNA Markers for the Diagnosis of Pancreatic and Biliary-Tract Cancers

It is difficult to detect pancreatic cancer or biliary-tract cancer at an early stage using current diagnostic technology. Utilizing microRNA (miRNA) markers that are stably present in peripheral blood, we aimed to identify pancreatic and biliary-tract cancers in patients. With “3D-Gene”, a highly sensitive microarray, we examined comprehensive miRNA expression profiles in 571 serum samples obtained from healthy patients, patients with pancreatic, biliary-tract, or other digestive cancers, and patients with non-malignant abnormalities in the pancreas or biliary tract. The samples were randomly divided into training and test cohorts, and candidate miRNA markers were independently evaluated. We found 81 miRNAs for pancreatic cancer and 66 miRNAs for biliary-tract cancer that showed statistically different expression compared with healthy controls. Among those markers, 55 miRNAs were common in both the pancreatic and biliary-tract cancer samples. The previously reported miR-125a-3p was one of the common markers; however, it was also expressed in other types of digestive-tract cancers, suggesting that it is not specific to cancer types. In order to discriminate the pancreato-biliary cancers from all other clinical conditions including the healthy controls, non-malignant abnormalities, and other types of cancers, we developed a diagnostic index using expression profiles of the 10 most significant miRNAs. A combination of eight miRNAs (miR-6075, miR-4294, miR-6880-5p, miR-6799-5p, miR-125a-3p, miR-4530, miR-6836-3p, and miR-4476) achieved a sensitivity, specificity, accuracy and AUC of 80.3%, 97.6%, 91.6% and 0.953, respectively. In contrast, CA19-9 and CEA gave sensitivities of 65.6% and 40.0%, specificities of 92.9% and 88.6%, and accuracies of 82.1% and 71.8%, respectively, in the same test cohort. This diagnostic index identified 18/21 operable pancreatic cancers and 38/48 operable biliary-tract cancers in the entire cohort. Our results suggest that the assessment of these miRNA markers is clinically valuable to identify patients with pancreato-biliary cancers who could benefit from surgical intervention.

Observation of the High-Tc Phase and Determination of the Pb Position in a Bi-Pb-Sr-Ca-Cu Oxide Superconductor

The high-Tc phase of a Bi-Pb-Sr-Ca-Cu oxide superconductor was observed directly by high-resolution transmission electron microscopy (HREM), and the atomic positions of Pb in the crystal were determined by high-resolution analytical electron microscopy (HRAEM) with a high spatial resolution 17 A using a probe 5 to 10 A in diameter. The HREM observation revealed that the crystal consisted solely of a triple Cu-O layered structure with c/2=18 A and without any intergrowth, and that the crystal structure was modulated along the b-axis. The HRAEM indicated that the Pb atoms were located in the Bi-O layers with an atomic ratio of Pb/Bi~0.1.

Formation of a 100 K Superconducting Bi(Pb)-Sr-Ca-Cu-O Film by a Spray Pyrolysis

A 100 K superconducting Bi(Pb)-Sr-Ca-Cu-O film was formed on a MgO(100) single crystal by a spray pyrolysis method. Fifteen hours heating of the as-sprayed film at 845°C in air was enough to give the superconducting film with a Tc zero higher than 100 K. An X-ray diffraction pattern showed that this film mainly consisted of the high-Tc phase with the orientation of the c-axis perpendicular to the surface.

Formation of as-deposited Y-Ba-Cu-O superconducting film by a high temperature spray pyrolysis method

An as-deposited Y-Ba-Cu-O superconducting film of which thickness was 7 µm was prepared by a spray pyrolysis method without post-annealing procedure. This film showed Tconset at 93 K and Tcend at 25 K. Its surface was smoother than that of a film heat treated at 950°C after the spraying process.

High Pressure Oxygen Treatment and the Substitution of Sr for Ba on (Nd1/3Ba2/3)2(Ce1/3Nd2/3)2Cu3Oy Superconductor

Substitution of Sr for Ba in the (Nd1/3Ba2/3)2(Ce1/3Nd2/3)2Cu3Oy superconductor has been performed together with high pressure oxygen treatment, and the relationship among Sr content, lattice parameter, hole concentration and superconducting properties has been clarified. It has been found that there is a strong relationship between superconducting transition temperature (Tc) and c-axis length. The highest Tconset and Tczero of the (Nd1/3(Ba3/4Sr1/4)2/3)2(Ce1/3Nd2/3)2Cu3Oy treated under 1300 atm oxygen pressure at 600°C for 24 hours are 59.3 K and 41.4 K, respectively. These transition temperatures are about 10 K higher than those of unsubstituted one.

A possible mechanism for hepatotoxicity induced by BIRB‐796, an orally active p38 mitogen‐activated protein kinase inhibitor

BIRB‐796, a selective inhibitor of p38 mitogen‐activated protein kinase, has entered clinical trials for the treatment of autoimmune diseases. Levels of alanine transaminase, a biomarker of hepatic toxicity in clinical pathology, were found to be increased in Crohns disease patients treated with BIRB‐796. The purpose of the present study was to clarify the molecular mechanism(s) of this hepatotoxicity. A toxicogenomic analysis using a highly sensitive DNA chip, 3D‐Gene™ Mouse Oligo chip 24k, indicated that BIRB‐796 treatment activated the nuclear factor (erythroid‐derived 2)‐like 2 signaling pathway, which plays a key role in the response to oxidative stress. A reactive intermediate of BIRB‐796 was detected by the glutathione‐trapping method using mouse and human liver microsomes. The production of this reactive metabolite in the liver may be one of the causes of BIRB‐796s hepatotoxicity. Copyright

An Ultrasensitive New DNA Microarray Chip Provides Gene Expression Profiles for Preoperative Esophageal Cancer Biopsies without RNA Amplification

Objectives: Gene expression profiling using pretreatment biopsies has been limited due to their small sample sizes. This study evaluated the usefulness of an ultrasensitive new DNA microarray chip, which has a unique array structure, for the clinical diagnosis of esophageal cancer using preoperative biopsies. Methods: Paired cancer and normal esophageal epithelial tissues from 56 patients who underwent esophagectomy and from 48 patients who underwent preoperative endoscopy were studied. Among 2 feature gene sets selected by a reference DNA chip discriminating malignant status of samples, 20 feature genes were selected for the development of the new DNA chip. The new DNA chip was hybridized with 0.1 µg of total RNA per slide without RNA amplification. Results: Twenty feature genes, including RRM-2 and XRCC-3, for the new DNA chip could discriminate cancer from noncancer at a 95.2% rate of accuracy in 42 biopsies (sensitivity 95.7%, specificity 94.7%). A receiver operating characteristic (ROC) curve analysis showed that the area under ROC curve for the prediction was 0.966. Conclusions: The gene expression profiles from the preoperative biopsies could diagnose esophageal cancer accurately, using the ultrasensitive DNA chip without RNA amplification. This new DNA chip technology might contribute further to the development of customized therapeutic strategies for various cancer patients.