bluuebunny/crossref_metadata_2025 huggingface dataset:
Frequent Allelic Loss at 7p14‐15 Associated with Aggressive Histologic Types of Breast Cancer
Authors: Kurose Keisuke , Iida Aritoshi , Araki Tsutomu , Sakamoto Goi , Kasumi Fujio , Nakamura Yusuke , Emi Mitsuru
Published: August 2005

We examined 142 primary human breast cancers to determine their patterns of loss of heterozygosity (LOH) at 19 microsatellite markers over the entire length of chromosome 7. Allelic loss at one or more loci on the short arm of chromosome 7 was observed in 37 of the tumors (26%). We found a new target region of allelic loss on 7p between D7S1802 and D7S817 at 7p14‐15. LOH on 7p was found more frequently in tumors of the invasive solid tubular and scirrhous type (31 of 87; 36%) than in other less aggressive types (2 of 27; 7%) (P=0.0047). The results suggest that inactivation of putative tumor suppressor gene(s) located at 7p14‐15 may play a role in the development and/or progression of primary breast cancers, particularly those of the invasive solid tubular and scirrhous type. Allelic loss was also found in 56 of 142 tumors on the long arm, and a commonly deleted region was defined between D7S522 and D7S1801 at 7q31.
Purification of Pierisin, an Inducer of Apoptosis in Human Gastric Carcinoma Cells, from Cabbage Butterfly, Pieris rapae
Authors: Watanabe Masahiko , Kono Takuo , Koyama Kotaro , Sugimura Takashi , Wakabayashi Keiji
Published: August 2005

A substance strongly cytotoxic to human carcinoma cell line TMK‐1 has been found in pupae, larvae and adults of the cabbage butterfly, Pieris rapae, and named pierisin. Pierisin was purified from the pupae of P. rapae by ammonium sulfate precipitation followed by DEAE‐cellulose, Phenyl‐Sepharose and hydroxyapatite column chromatographies. The molecular weight of the purified pierisin, which was homogeneous on SDS‐polyacrylamide gel, was analyzed by mass spectrometry and found to be 98 kDa. Pierisin showed a strong cytotoxic effect, with an IC50 of 0.75 ng/ml for human gastric carcinoma TMK‐1 cells. The dying cells exhibited characteristic morphological features of apoptosis, such as cell shrinkage, chromatin condensation and nuclear fragmentation. Oligonucleosomal DNA fragmentation was also observed in DNA isolated from pierisin‐treated cells. Moreover, similar characteristic changes showing apoptotic cell death were observed in TMK‐1 cells treated with a crude extract of pupae of P. rapae. These results indicate that pierisin from the pupae of P. rapae induces apoptosis in human carcinoma cells.
Intracellular Signal‐transducing Elements Involved in Transendothelial Migration of Lymphoma Cells
Authors: Tsuzuki Shinobu , Toyama‐Sorimachi Noriko , Kitamura Fujiko , Tsuboi Hiroko , Ando Joji , Sakurai Takashi , Morii Narito , Narumiya Shuh , Miyasaka Masayuki
Published: August 2005

To investigate the molecular mechanisms underlying transendothelial migration of tumor cells, an essential process for their hematogenous dissemination, we developed an in vitro model system that allows the separate monitoring of cell adhesion and transmigration processes. This system uses a human pre‐B lymphoma cell line, Nalm‐6, and a cultured mouse endothelial cell line, KOP2.16. Nalm‐6 cells rapidly adhered to KOP2.16 and subsequently transmigrated underneath them. Using this model, we examined the effects on transendothelial migration, of various reagents which specifically interfere with the function of intracellular signal transduction molecules. Treatment of Nalm‐6 cells with wortmannin (WMN), herbimycin A, pertussis toxin, or C3 exoenzyme of Clostridium botulinum, which specifically inhibit P13 kinase and/or myosin light chain kinase, herbimycin‐sensitive tyrosine kinases, heterotrimeric G proteins, and the small G proteins rho/rac, respectively, reduced transmigration in a dose‐dependent manner. Pretreatment of KOP2.16 endothelial cells with WMN also reduced transmigration in a dose‐dependent manner. Binding of Nalm‐6 to KOP2.16 was not affected, even when Nalm‐6 or KOP2.16 cells were pretreated with these inhibitors, indicating that the reduction of transmigration was not due to a reduction of Nalm‐6 binding to KOP2.16. These results also indicate that the signal transduction pathway(s) involved in transmigration can be dissociated from that of adhesion. Our results support the notion that endothelial cells are not a passive barrier in lymphoma extravasation, but that they assist lymphoma cell extravasation.
Effect of Gelonin Immunoconjugate with Monoclonal Antibody MSN‐1 to Endometrial Adenocarcinoma on Antigen‐producing Tumor Cells in vivo
Authors: Kaneta Yoshibumi , Tsukazaki Katsumi , Kubushiro Kaneyuki , Aoki Rui , Sakayori Motoko , Ueda Masakazu , Nozawa Shiro
Published: August 2005

Missile therapy, which destroys cancer cells specifically, has been advocated as an effective modality for the treatment of carcinoma. We have developed an immunoconjugate consisting of the monoclonal antibody MSN‐1 (IgM), which reacts strongly with endometrial adenocarcinomas, combined with a plant hemitoxin named gelonin via a disulfide bond using N‐succinimidyl‐3‐(2‐pyridyldithio)propionate and 2‐iminothiolane, and examined its selective cytotoxicity in athymic mice. The reductions in resected weights of target tumor cells, at the local site of MSN‐1‐gelonin immunoconjugate treatment, were 96% with local administration and 75% with caudal vein administration, as compared with the untreated group. There was no weight loss in treated mice. Our results suggest that this MSN‐1‐gelonin immunoconjugate has potential clinical applications in the treatment of endometrial adenocarcinomas.
Effect of Gene Transfer of Tumor Necrosis Factor Receptors into Human Lung Carcinoma Cell Line
Authors: Ohara Hisao , Hasegawa Yoshinori , Kawabe Tsutomu , Ichiyama Satoshi , Hara Toru , Shimono Yohei , Saito Hidehiko , Shimokata Kaoru
Published: August 2005

The human lung adenocarcinoma cell line A549 is known to be resistant to tumor necrosis factor alpha (TNF‐α)‐mediated tumor cell lysis in spite of the expression of 55 kDa TNF receptor (TNF‐R55) mRNA and its cell surface protein. In this study, we investigated the mechanism of TNF‐α resistance and the role of two types of TNF receptors (TNF‐R55 and TNF‐R75 (75 kDa TNF receptor)). TNF‐R55 or TNF‐R75 cDNA was transfected into A549 cells. In addition, a C‐terminal deletion mutant of TNF‐R75 which lacks the intracellular domain of TNF‐R75 was also transfected into A549 cells. We assessed the TNF‐α‐mediated tumor cell lysis of these transfected clones, and found that the cytotoxic effect increased in transfected clones highly expressing TNF‐R55, but not in low‐expression clones. As for TNF‐R75, the cytotoxic effect of TNF‐α was observed in TNF‐R75‐transfected clones even when expression was low. Furthermore, the cytotoxic effect was also observed in clones transfected with the deletion mutant of TNF‐R75, as well as the complete TNF‐R75. These results indicate that a certain level of expression of TNF‐R55 is necessary for obtaining TNF‐α‐mediated tumor cell lysis in the absence of TNF‐R75. On the other hand, the expression of TNF‐R75 strongly induces TNF‐α‐mediated cytotoxicity through TNF‐R55 in the absence of an intracellular signal via TNF‐R75.
Trends of Lung Cancer Incidence by Histologic Type: A Population‐based Study in Osaka, Japan
Authors: Sobue Tomotaka , Ajiki Wakiko , Tsukuma Hideaki , Oshima Akira , Hanai Aya , Fujimoto Isaburo
Published: August 2005

We investigated trends of lung cancer incidence from 1974 to 1993 by histologic type, using data from the population‐based cancer registry in Osaka, Japan. Since the proportion of cases with histologic types identified was not sufficiently high, sex‐ and age‐specific incidence rates by histologic types were estimated assuming that the distribution of histologic types was the same across the same sex and age group regardless of reporting status. Cumulative risk from 0 to 74 years old for total lung cancer increased 1.3‐fold from the period 1974–77 to 1986–89 and then plateaued in the period 1990–93 for both males and females. When divided into histologic types, cumulative risk for incidence of squamous cell carcinoma was almost constant during the study period for both males and females. During the same period, adenocarcinoma increased up to 1.4‐fold for both males and females. This increase seemed to have reached a plateau recently for males, but not for females. Small cell carcinoma increased monotonously up to 1.6‐ to 1.7‐fold for both males and females. Large cell carcinoma showed over 2‐fold increase for both males and females; however, the estimates fluctuated due to the small number of cases. This study provides further evidence of a relative increase of adenocarcinoma compared to squamous cell carcinoma. Recent trends of tapering increase of lung cancer incidence should be confirmed by further observation.
Analysis of the c‐myc, K‐ras and p53 Genes in Methylcholanthrene induced Mouse Sarcomas
Authors: Watanabe Hiroshi , Shimokado Kiyoshi , Asahara Toshimasa , Dohi Kiyohiko , Niwa Ohtsura
Published: August 2005

We have examined 63 methylcholanthrene (MCA)‐induced mouse sarcomas for possible correlations of mutations involving the c‐myc, ras and p53 genes. The c‐myc gene was found to be amplified in 18 of these sarcomas (29%). Polymerase chain reaction‐single strand conformation polymorphism (PCR‐SSCP) analysis and subsequent direct sequencing identified 18 cases carrying K‐ras mutation at codons 12, 13 and 61 (29%). No mutation was detected in the H‐ras and N‐ras genes. Mutations of the p53 gene in exons 5 to 8 were found in 45 cases (71%). Comparison of these mutations revealed that out of 18 cases with c‐myc gene amplifications, 10 carried K‐ras mutations (56%) and 14 carried p53 mutations (78%). In contrast, among 45 cases of sarcomas without c‐myc gene amplification, 8 were found to have K‐ras mutations (18%). The same 45 cases were found to have 31 p53 mutations (69%). The present study suggests a strong correlation between c‐myc gene amplification and K‐ras gene mutation (P < 0.01). p53 gene mutation was frequently found among MCA‐induced mouse sarcomas, indicating the importance of this mutation in the etiology of these tumors. However, p53 mutations were present in sarcomas regardless of the state of c‐myc amplification and K‐ras mutation. Therefore, a defect in the p53 gene is independent of amplification of the c‐myc gene or point mutation of the K‐ras gene.
Natural Killer Activity in a Medium‐term Multi‐organ Bioassay for Carcinogenesis
Authors: Tozzi Spinardi Ana Lúcia , Kaneno Ramon , Rodrigues Maria Aparecida Marchesan , Salvadori Daisy Maria Fávero , Rocha Noeme Sousa , Barbisan Luís Fernando , Ribeiro Lúcia Regina , de Camargo João Lauro Viana
Published: August 2005

Natural killer (NK) cell activity was evaluated after the initiation and promotion steps in a medium‐term multi‐organ bioassay for carcinogenesis. NK cell activity was assessed in vitro by Cr51 release assay at the 4th and 30th weeks of the experiment. Male Wistar rats were sequentially initiated with N‐diethylnitrosamine (DEN i.p.), N‐butyl‐N‐(4‐hydroxybutyl)nitrosamine (BBN drinking water), N‐methyl‐N‐nitrosourea (MNU i.p.), dihydroxy‐di‐N‐propylnitrosamine (DHPN drinking water) and N, N′‐dimethylhydrazine (DMH s.c.) at subcarcinogenic doses for 4 weeks (DMBDD initiation). One group was evaluated at the 4th week and the other was maintained without any further treatment until the 30th week. Two initiated groups were exposed through the diet to 2‐acetylaminofluorene (2‐AAF) or phenobarbital (PB), from the 6th until the 30th week. Five additional groups were studied to evaluate the effects of each initiator on NK activity. All groups submitted to initiation only, initiation plus promotion, or promotion only, developed significantly more preneoplastic lesions than the untreated control group. The main target organs for tumor development in the initiated animals were the liver and the colon, irrespective of treatment with 2‐AAF or PB. NK cell activity was not affected by exposure to genotoxic carcinogens after initiation, at the 4th week. Treatments only with PB or 2‐AAF did not change NK cell activity. However, decreased NK cell activity was registered in the group only initiated with DMBDD and in the group given DMBDD+2‐AAF. This late depression of NK cell activity at the 30th week could be related to the production of suppressing molecules by the tumor cells.
Enhancement of Cisplatin Sensitivity in High Mobility Group 2 cDNA‐transfected Human Lung Cancer Cells
Authors: Arioka Hitoshi , Nishio Kazuto , Ishida Tomoyuki , Fukumoto Hisaoh , Fukuoka Kazuya , Nomoto Taisuke , Kurokawa Hirokazu , Yokote Hideyuki , Abe Shosaku , Saijo Nagahiro
Published: August 2005

To elucidate the role of high mobility group 2 protein (HMG2) in cis‐diamminedichloroplatinum (II) (cisplatin, CDDP) sensitivity, we constructed a human HMG2‐transfected human non‐small cell lung cancer cell line, PC‐14/HMG2. The HMG2 mRNA expression level was approximately twice those of parental PC‐14 and mock‐transfected PC‐14/CMV. Gel mobility shift assay revealed a CDDP‐treated DNA‐protein complex in the nuclear extract of PC‐14/HMG2, which was not found in the extracts of PC‐14 and PC‐14/CMV. This complex formation was subject to competition by CDDP‐treated non‐specific salmon sperm DNA, indicating that ectopic HMG2 recognizes CDDP‐damaged DNA. PC‐14/HMG2 showed more than 3‐fold higher sensitivity to CDDP than PC‐14 and PC‐14/CMV. The intracellular platinum content of PC‐14/HMG2 after exposure to 300 μM CDDP was 1.1 and 1.5 times that of PC‐14 and PC‐14/CMV, respectively. Cellular glutathione levels were not different in these cell lines. Repair of DNA interstrand cross‐links determined by alkaline elution assay was decreased in PC‐14/HMG2. These results suggest that HMG2 may enhance the CDDP sensitivity of cells by inhibiting repair of the DNA lesion induced by CDDP.
Reduction of the Side Effects of an Antitumor Agent, KRN5500, by Incorporation of the Drug into Polymeric Micelles
Authors: Matsumura Yasuhiro , Yokoyama Masayuki , Kataoka Kazunori , Okano Teruo , Sakurai Yasuhisa , Kawaguchi Takanori , Kakizoe Tadao
Published: August 2005

For intravenous (i.v.) injection of a water‐insoluble antitumor drug, KRN5500, we have successfully incorporated KRN5500 into polymeric micelles. In the present study, in vitro and in vivo antitumor activity against several human tumor cell lines and toxicity in mice of polymeric micelles incorporating KRN5500 (KRN/m) were evaluated in comparison with those of the prototype KRN5500. KRN/m was found to express similar antitumor activity to KRN5500 in the in vitro and in vivo systems. However, the vascular damage and liver focal necrosis observed with KRN5500 i.v. injection were not seen when KRN/m was administered i.v. Therefore, we expect that KRN/m will be superior to KRN5500 for clinical use and that the methodology of polymeric micelle drug carrier systems can be applied to other water‐insoluble drugs.
Screening for Nicotine Dependence among Smoking‐related Cancer Patients
Authors: Mikami Ichiro , Akechi Tatsuo , Kugaya Akira , Okuyama Toru , Nakano Tomohito , Okamura Hitoshi , Yamawaki Shigeto , Uchitomi Yosuke
Published: August 2005

To identify lung and head‐and‐neck cancer patients who will have difficulty stopping smoking it is necessary to measure the severity of their nicotine dependence. In this study, we compiled a Japanese version of the Fagerström test for nicotine dependence (FTND) and examined its reliability and validity. One hundred and fifty‐one cancer patients participated in this study and took our Japanese version of the FTND. Socio‐demographic and medical data and information about smoking habits were obtained from a semi‐structured interview, and the patients' nicotine dependence was evaluated according to the Diagnostic and Statistical Manual of Mental Disorders, 3rd Ed., Rev. (DSM–III–R). The mean FTND scores±SD of the group with nicotine dependence and the group without nicotine dependence were 6.85±2.00 and 3.70±2.13 respectively, and the difference was significant (P < 0.001, Mann‐Whitney's U‐test). The test‐retest correlation was 0.75. Cronbach's a of the FTND was 0.66. The FTND score correlated significantly with the number of satisfied criteria of nicotine dependence (r=0.70; P <0.001, Pearson's correlation). By using a receiver‐operating‐characteristic curve, we determined a score of 5/6 as a suitable cut‐off point for nicotine dependence; this point gave high sensitivity and specificity (0.75 and 0.80, respectively). These results suggest that our Japanese version of FTND is a reliable and valid measure of nicotine dependence in patients with smoking‐related cancers.
Relationship between Multicentric Occurrence of Hepatocellular Carcinoma and Histology of Noncancerous Hepatic Tissue in Patients with Chronic Hepatitis C
Authors: Kubo Shoji , Yamamoto Takatsugu , Ikebe Takashi , Shuto Taichi , Hirohashi Kazuhiro , Tanaka Hiromu , Tsukamoto Tadashi , Wakasa Kenichi , Kinoshita Hiroaki
Published: August 2005

The relationship between multicentric occurrence of hepatocellular carcinoma (HCC) and the histology of noncancerous hepatic tissue was investigated in 252 patients infected with hepatitis C virus (HCV) and surgically treated for HCC. One type of multicentric HCC had at least one tumor consisting of well‐differentiated HCC, together with moderately or poorly differentiated HCC located in a separate region. The other type had an area of well‐differentiated component around HCC with less differentiation in all occurrences. Noncancerous hepatic tissues were assessed using a histologic activity index score. Serum alanine aminotransferase (ALT) activity, the concentration of type 4 collagen, the grading score (severity of active hepatitis), and the staging score (degree of fibrosis) were significantly higher in patients with multicentric HCCs than in those without them. Platelet count was significantly lower in patients with multicentric HCCs. The prevalence of multicentric HCCs increased as the grading score and staging score increased. On univariate analysis, a low platelet count and high grading and staging scores were risk factors for multicentric HCCs. A high ALT activity and a high concentration of type 4 collagen tended to be risk factors. On multivariate analysis, high grading score and high staging score were independent risk factors. These findings indicate that active hepatitis and extensive fibrosis are responsible for the development of multicentric HCCs. Measurement of platelet count, ALT activity, and the concentration of type 4 collagen, and histologic assessment of noncancerous hepatic tissue provide information useful for estimation of the potential for multicentric carcinogenesis.
Genetic Alterations in Ulcerative Colitis‐associated Neoplasia Focusing on APC, K‐ras Gene and Microsatellite Instability
Authors: Umetani Naoyuki , Sasaki Shin , Watanabe Toshiaki , Shinozaki Masaru , Matsuda Keiji , Ishigami Hironori , Ueda Eiji , Muto Tetsuichiro
Published: August 2005

The status of genetic alterations in ulcerative colitis (UC)‐associated neoplasia (UCAN) was investigated focusing on microsatellite instability (MSI) which is seen in a certain fraction of colorectal carcinomas, and adenomatous polyposis coli (APC) gene and K‐ras gene, in which mutations occur in the early stage of sporadic colorectal tumorigenesis. Thirty‐one UCAN from 15 UC patients who had undergone colorectal resection at our institution were investigated. There were 8 lesions of invasive carcinoma, 15 high‐grade dysplasia (HGD) and 8 low‐grade dysplasia (LGD). DNA was extracted from each neoplastic lesion and corresponding non‐neoplastic tissue by a microdissection method. MSI status at 9 microsatellite loci, loss of heterozygosity (LOH) at the APC locus, and K‐ras codon 12 point mutation were examined. As for MSI, 4/31 (13%) UCAN (carcinoma: 1/8 (13%), HGD: 2/15 (13%), LGD: 1/8 (13%)) were MSI‐high (3 or more unstable loci) and 12/31 (39%) UCAN (carcinoma: 3/8 (38%), HGD: 6/15 (40%), LGD: 3/8 (38%)) were MSI‐low (1 or 2 unstable loci). LOH at the APC locus was not found in 9 UCAN from 6 informative (heterozygous) cases. The K‐ras mutation rate of UCAN was 3/31 (9.7%) (carcinoma: 2/8 (25%), HGD: 1/15 (7%) and LGD: 0/8). MSI is relatively common in UCAN and is present at the early stage of tumorigenesis of UCAN, while the involvement of genetic alterations of the APC gene and K‐ras gene is small. MSI may be one of the mechanisms of the increased neoplastic risk in UC, and UCAN may develop through a different carcinogenic pathway from sporadic carcinomas.
Analysis of a Chronic Myelogenous Leukemia Patient Vaccinated with Leukemic Dendritic Cells Following Autologous Peripheral Blood Stem Cell Transplantation
Authors: Fujii Shin‐ichiro , Shimizu Kanako , Fujimoto Koji , Kiyokawa Tetsuyuki , Shimomura Taizo , Taniguchi Osamu , Kinoshita Moritoshi , Kawano Fumio
Published: August 2005

Dendritic cells (DCs) are believed to be the most potent antigen‐presenting cells and may be important in the induction of anti‐leukemia specific T cell responses. In this preliminary clinical study, a patient with chronic phase chronic myelogenous leukemia (CML) was vaccinated with autologous leukemic DCs following autologous peripheral blood stem cell transplantation (PBSCT). In an in vitro study, leukemic DCs were generated using granulocyte‐macrophage colony‐stimulating factor (GM‐CSF), tumor necrosis factor‐α, and interleukin‐4 from granulocyte colony‐stimulating factor (G‐CSF)‐mobilized PBSC fraction of this patient, and were found to be Phl+, and to possess the morphologic and phenotypic characteristics of mature DCs. These cells could also elicit antigen‐specific immune responses, including a vigorous cytotoxicity specific to CML cells. In the clinical experiment, we obtained evidence that infused leukemic DCs could induce T cell clones expressing the same T cell receptor usage as a cytotoxic T cell line, suggesting that the immune repertoire includes tumor‐reactive T cells. These cytotoxic T lymphocytes are activated in vivo. The vaccination of leukemic DC caused a decrease in the number of Phl+ cells in the peripheral blood and bone marrow. These results indicate that the activity is an immunologically mediated phenomenon and vaccination therapy with leukemic DC following autologous PBSCT may be effective in treating CML.
Effectiveness of a Simply Designed Tumor Vaccine in Prevention of Malignant Melanoma Development
Authors: Novakovic Srdjan , Ihan Alojz , Jezereks Barbara
Published: August 2005

We investigated the efficacy of a simple syngeneic tumor vaccine to induce specific antitumor immunity in female C57B1/6 mice. Tumor vaccine was prepared by mixing irradiated B‐16 melanoma tumor cells with the pleiotropic biological response modifier—maleic anhydride divinyl ether (MVE‐2). Experimental animals were pretreated with the vaccine in order to prevent the development of intraperitoneal (i.p.) B‐16 melanoma tumors after inoculation of viable tumor cells. More than 40% of prevaccinated animals challenged i.p. with 5×105 viable tumor cells were completely protected from tumor development and remained tumor‐free 100 days after tumor cell inoculation. The percentage of tumor‐free animals (survivors) rose to as much as 90% when the application of tumor vaccine was repeated two weeks after the first vaccination (i.e. one week after the inoculation of viable tumor cells). The induced antitumor response depended predominantly upon macro‐phage function, since vaccinated animals which were depleted of peritoneal macrophages died within the same time range as animals in the control group. Also, tumor‐type specificity of the vaccine was confirmed by the fact that the animals vaccinated with B‐16 melanoma vaccine were not protected from the development of another type of tumor. In conclusion, comparison of the experimental data with the data from the literature suggests that our simple tumor vaccine may be as effective as genetically engineered tumor vaccines. At the same time, this kind of vaccine is easier to control and thus safer to apply in humans when compared to genetically engineered vaccines.
Enhanced Sonodynamic Antitumor Effect of Ultrasound in the Presence of Nonsteroidal Anti‐inflammatory Drugs
Authors: Sakusabe Noboru , Okada Kyoji , Sato Kozo , Kamada Shinko , Yoshida Yasuo , Suzuki Toshio
Published: August 2005

The antitumor effects of non‐steroidal anti‐inflammatory drugs, tenoxicam and piroxicam, against sarcoma 180 cells cultured in 7‐week‐old male mice were examined in vitro under ultrasonic irradiation. The survival rate of tumor cells when tenoxicam or piroxicam was added to sarcoma 180 suspension under ultrasonic irradiation was significantly lower than that when ultrasound alone was applied. Furthermore, when L‐histidine, a scavenger of singlet oxygen and hydroxyl radical, or D‐mannitol, a scavenger of hydroxyl radical, was used concurrently, the survival rate of tumor cells was significantly higher with L‐histidine. From the above findings, it is surmised that tenoxicam and piroxicam increase the antitumor effects of ultrasound by increasing the production of singlet oxygen and other active oxygen species.
A Selective Cyclooxygenase‐2 Inhibitor Suppresses Tumor Growth in Nude Mouse Xenografted with Human Head and Neck Squamous Carcinoma Cells
Authors: Nishimura Goshi , Yanoma Syunsuke , Mizuno Hiromi , Kawakami Koji , Tsukuda Mamoru
Published: August 2005

The anti‐tumor effect of a selective cyclooxygenase (COX)‐2 inhibitor, JTE‐522, was examined with the human head and neck squamous cell carcinoma cell line KB. KB cells do not produce prostaglandin (PG)‐E2. In vitro, JTE‐522 induced an increase of G1 phase‐arrested cells, suppression of platelet‐derived growth factor (PDGF) production and inhibition of telomerase activity. No cytotoxic effect was detected. In vivo, the growth of the tumor xenografted into nude mice was significantly suppressed by JTE‐522. Suppression of angiogenesis at the periphery of the tumor, increase of G1‐arrested cells and suppression of telomerase activity were observed, together with an increase of apoptotic cell death in the tumor. Immunological enhancement did not play a role. We concluded that the anti‐tumor effect of JTE‐522 was caused by anti‐angiogenesis action, cell cycle arrest and inhibition of telomerase activity of the tumor cells. These combined effects might induce apoptosis.
Preoperative Clinical Radioimmunodetection of Pancreatic Cancer by 111In‐labeled Chimeric Monoclonal Antibody Nd2
Authors: Sawada Tetsuji , Nishihara Tamahiro , Yamamoto Atsushi , Teraoka Hitoshi , Yamashita Yoshito , Okamura Terue , Ochi Hironobu , Ho Jenny J. L. , Kim Young‐Sik , Hirakawa Kosei
Published: August 2005

The present study was carried out with the purpose of evaluating the clinical usefulness of radioimmunodetection (RAID) with 111In‐labeled murine/human chimeric monoclonal antibody, Nd2 (c‐Nd2) in patients with pancreatic cancer. Nineteen patients suspected to have pancreatic cancer were administered intravenously 74 MBq/2 mg 111In‐labeled c‐Nd2 in 100 ml of saline containing 2% albumin over 30 min. A scintigram was obtained on the 3rd day after infusion by using single photon emission computed tomography (SPECT) imaging. Of the 14 patients finally diagnosed as having pancreatic cancer on the basis of surgical specimens or progress of disease, specific focal uptake at the site of the tumor was detected in 12 (true positive cases), representing a sensitivity of 85.7% (12/14), and liver metastasis was found in one case with metastasis. Of the 5 patients diagnosed with tumor‐forming pancreatitis (TFP), 4 patients demonstrated true negative imaging, but one patient whose tumor demonstrated interesting findings in histology and immunostaining, showed false positive imaging. Of patients investigated for human anti‐chimeric antibody (HACA) response, none showed HACA response, and no allergic reaction was seen in any of the patients administered c‐Nd2. These results suggest that RAID with 111In‐labeled c‐Nd2 is useful for differential preoperative diagnosis between invasive pancreatic cancer and TFP.
Mixed Phenotype Lymphomas in Thymectomized (SL/KhxAKR/Ms)F1 Mice
Authors: Lu Ling‐Min , Hiai Hiroshi
Published: August 2005

Lymphomagenesis in mice is determined both by genetic and epigenetic mechanisms. The inbred strain SL/Kh mice selectively develop pre‐B lymphomas and AKR/Ms, T‐lymphomas. In crosses between SL/Kh and AKR/Ms, an AKR‐derived dominant gene Tlsm1 (Thymic lymphoma susceptible mouse‐1) determines the type of lymphoma to be a T‐lymphoma. As an approach to the role of Tlsm1, we studied the effect of thymectomy at 1 week of age in (SL/KhxAKR/Ms)Fl hybrids. In intact F1 mice, the predominant type of lymphoma was of T‐lineage, whereas in thymectomized mice, it was an unusual mixed‐phenotype lymphoma. They were basically CD5+ B‐lymphomas with a rearranged immunoglobulin gene, but carried NK1 and Mac1 on the cell surface and large lysosomal granules in the cytoplasm. Histologically, the lymphoma consisted of large lymphoblastoid cells and infiltrated the spleen, lymph node and liver. Electron microscopy and histochemistry revealed numerous cytoplasmic granules containing acid phosphatase and lysozyme. These morphological features are suggestive of large granular lymphocytes. They expressed interleukin‐4, perforin, and interferon‐γ. On transplantation, these lymphoma cells grew equally well in intact and thymectomized F1 recipients.
Somatic Mutations in RET Exons 12 and 15 in Sporadic Medullary Thyroid Carcinomas: Different Spectrum of Mutations in Sporadic Type from Hereditary Type
Authors: Uchino Shinya , Noguchi Shiro , Yamashita Hiroto , Sato Mari , Adachi Mitsuo , Yamashita Hiroyuki , Watanabe Shin , Ohshima Akira , Mitsuyama Shoshu , Iwashita Toshihide , Takahashi Masahide
Published: August 2005

Germline mutations in the RET proto‐oncogene are responsible for multiple endocrine neoplasia type 2 (MEN 2A and 2B) and familial medullary thyroid carcinoma (FMTC). Point mutations or in‐frame deletions of exons 10, 11, 13, 14 and 16 are associated with sporadic medullary thyroid carcinoma (MTC). To understand further the role of the RET gene in sporadic MTC, we examined mutations in exons 12 and 15 of RET in patients with sporadic MTC. DNAs were extracted from 39 formalin‐fixed tumor tissues and corresponding normal thyroid tissues or peripheral blood leukocytes. DNA sequencing was used to identify mutations in exons 12 and 15 of RET. In this study, one novel somatic mutation was found in exon 12 and five novel mutations or deletions were found in exon 15. Of the patients with mutations, one had an in‐frame 12‐bp deletion (nt. 2625‐2636), one had point mutations in both codons 884 and 908, and the remaining three had point mutations in codons 748, 876 and 901, respectively. Together with our previous identification of somatic mutations in exons 10, 11, 13, 14 and 16, somatic alterations were found in 10 out of 39 (25.6%) sporadic MTCs. There was no association of RET gene mutations with tumor recurrence or prognosis. These results suggest that mutations occur frequently in the RET coding region in addition to the previously reported mutation hot spots, and there is a different spectrum of mutations between sporadic and hereditary MTC.
Clonal Analysis of Adenosquamous Carcinoma of the Lung
Authors: Niho Seiji , Yokose Tomoyuki , Kodama Tetsuro , Nishiwaki Yutaka , Mukai Kiyoshi
Published: August 2005

Adenosquamous carcinoma of the lung is a subset of pulmonary carcinomas, and comprises less than 4% of lung carcinomas. Its histogenesis remains unclear. The clonality of adenosquamous carcinoma from four female patients was analyzed to determine whether the clonality between the squamous cell and adenocarcinomatous components coincides. Each lesion was precisely microdissected from methanol‐fixed sections. Adjacent normal lung tissue was collected as a normal control. DNA was extracted for clonal analysis based on an X‐chromosome‐linked polymorphic marker, the human androgen receptor gene (HUMARA). HUMARA was found to be amplified with or without previous digestion by the methylation‐sensitive restriction endonuclease HpaII. All four cases were informative. Squamous cell and adenocarcinomatous components showed identical monoclonal patterns in all four patients. In one case, only the squamous cell carcinomatous component showed loss of heterozygosity of the HUMARA locus. The results suggest that the squamous cell and adenocarcinomatous components originate from the same cell.
TAC‐101 (4‐[3,5‐bis(trimethylsilyl)benzamido]benzoic acid) Inhibits Spontaneous Mediastinal Lymph Node Metastasis Produced by Orthotopic Implantation of Lewis Lung Carcinoma
Authors: Murakami Koji , Yamaura Takeshi , Suda Kazuhito , Ohie Shinji , Shibata Jiro , Toko Toshiyuki , Yamada Yuji , Saiki Ikuo
Published: August 2005

The anti‐tumor and anti‐metastatic effects of 4‐[3,5‐bis(trimethylsilyl)benzamido]benzoic acid (TAC‐101) were investigated using our established lung cancer model. Orthotopic implantation of Lewis lung carcinoma (LLC) cells into the lung parenchyma produced a solitary tumor nodule in the lung followed by mediastinal lymph node metastasis. Daily oral administration of TAC‐101 at doses ranging from 4 to 16 mg/kg resulted in a significant inhibition of lymphatic metastasis (inhibition rate=57 to 76%), while only the dose of 16 mg/kg significantly inhibited tumor growth at the implanted sites (inhibition rate=46%). Combined treatment with cis‐diamminedichloroplatinum (CDDP) and TAC‐101 (8 mg/kg, p.o., daily) enhanced the anti‐tumor effect of CDDP (7 mg/ kg, i.v., bolus) against both the growth of implanted tumor and lymphatic metastasis. In addition, this combined treatment significantly prolonged the survival time of LLC tumor‐bearing mice as compared to treatment with each agent alone. The anti‐activating protein‐1 (AP‐1) activity of TAC‐101 caused inhibition of LLC cell invasion through the repression of expression of urokinase‐type plasminogen activator and its receptor. The anti‐invasive activity of TAC‐101 may be involved in its in vivo anti‐metastatic activity. These findings suggest that TAC‐101 is a novel anti‐cancer agent that may improve the therapeutic modalities for lung cancer patients with metastatic disease.
Epstein‐Barr Virus‐encoded Latent Membrane Protein 1 Co‐expresses with Epidermal Growth Factor Receptor in Nasopharyngeal Carcinoma
Authors: Sheen Tzung‐Shiahn , Huang Yu‐Tzu , Chang Yih‐Leong , Ko Jenq‐Yuh , Wu Chung‐Shun , Yu Yu‐Chen , Tsai Ching‐Hwa , Hsu Mow‐Ming
Published: August 2005

Latent membrane protein 1 (LMP‐1) is the only Epstein‐Barr virus (EBV)‐encoded oncogenic protein that has been detected in nasopharyngeal carcinoma (NPC), a cancer that is closely associated with EBV. Previous in‐vitro studies have demonstrated that LMP‐1 can upregulate epidermal growth factor receptor (EGFR) in epithelial cells. It was not established whether this cellular effect exists in NPC. To assess the association between LMP‐1 and EGFR in NPC tissues, 60 NPC specimens were examined by immunohistochemistry using anti‐LMP‐1 antibody (CS 1–4) and anti‐EGFR antibodies (EGFR 1, EGFR 1005). The results revealed that 41 (68.3%) specimens were immunopositive for LMP‐1 and 44 (73.3%) specimens over‐expressed EGFR. Morphologically, the expressions of LMP‐1 and EGFR were homogeneously distributed in the tumor nests. In addition, the correlation between LMP‐1 and EGFR was statistically significant (P<0.001, χ2 test, d.f.=1). To elucidate further the correlation between LMP‐1 and EGFR in vivo and in situ, an indirect dual immunofluorescence assay was conducted, using secondary antibodies conjugated with fluorescein isothiocyanate (FITC) or indocarbocyanine (Cy3). The results disclosed an intimate co‐expression of LMP‐1 and EGFR. In summary, the data indicate that over‐expression of EGFR is a common phenomenon in NPC, and that EGFR is co‐expressed with LMP‐1 in NPC. Thus, EBV may play a role in the tumorigenesis of NPC through the effects of LMP‐1 and EGFR.
Correlation of Clinicopathologic Features of Resected Hepatocellular Carcinoma with Hepatitis C Virus Genotype
Authors: Murase Junya , Kubo Shoji , Nishiguchi Shuhei , Hirohashi Kazuhiro , Shuto Taichi , Ikebe Takashi , Kinoshita Hiroaki
Published: August 2005

Clinicopathologic findings in patients with hepatocellular carcinoma complicating hepatitis C virus and outcomes after liver resection were compared between different viral genotypes. One hundred and forty‐seven patients with both anti‐hepatitis C virus antibody and hepatitis C virus RNA in their sera underwent curative resection for hepatocellular carcinoma in our department between 1991 and 1997. Of these patients, 115 were infected with hepatitis C virus genotype 1b (group 1), and 32 were infected with 2a or 2b (group 2). Clinicopathologic findings and outcomes after operation were compared between the two groups. Alanine aminotransferase activity was significantly higher in group 2 than in group 1. Genotypes did not differ concomitantly with histopathologic features of the carcinoma or adjacent hepatic tissue. Although the tumor‐free survival rate did not differ significantly between the two groups, recurrence was not detected during the period beyond 3 years following operation in group 2, while recurrences arose during that period in 16 group 1 patients, most of whom continued to manifest active hepatitis. In 7 of these 16 patients, the recurrent tumors were histologically multicentric in origin. The cumulative survival rate was significantly lower in group 1 than 2. Multivariate analysis indicated that genotype 1b was an independent risk factor for short survival. Patients infected with genotype 1b may have a relatively high risk of ongoing hepatocarcinogenesis and more aggressive progression of associated liver dysfunction, resulting in a poorer outcome than with other genotypes.
Infrequent BCL10 Mutations in B‐Cell Non‐Hodgkin's Lymphomas
Authors: Takahashi Hidenobu , Hosokawa Yoshitaka , Suzuki Ritsuro , Morishima Yasuo , Nakamura Shigeo , Seto Masao
Published: August 2005

The BCL10 gene was recently isolated from the breakpoint region of t(1;14)(p22;q32) in mucosa‐associated lymphoid tissue (MALT) lymphomas. Somatic mutations of BCL10 were found in not only t(1;14)‐bearing MALT lymphomas, but also a wide range of other tumors. To clarify the actual frequency and spectrum of BCL10 mutations in primary B‐cell non‐Hodgkin's lymphomas (NHL), we examined a total of 139 NHL cases comprising 25 with MALT lymphomas, 54 with follicular B‐cell lymphomas (FCL), and 60 with diffuse large B‐cell lymphomas (DLBL). Polymerase chain reaction single‐strand conformation polymorphism (PCR‐SSCP) and sequencing analyses led to the identification of four nucleotide changes in FCL and one in DLBL. In contrast, no BCL10 mutations were found in our series of MALT lymphomas. While screening for mutations, we also found three polymorphic sequence variants at codons 5 and 213 and in intron 1 of the BCL10 gene. Our results strongly suggest that somatic mutations of BCL10, if they occur at all, are rare in B‐cell NHLs and do not commonly contribute to their molecular pathogenesis.
G1–checkpoint Function Including a Cyclin‐dependent Kinase 2 Regulatory Pathway as Potential Determinant of 7–Hydroxystaurosporine (UCN‐01)‐induced Apoptosis and G1–phase Accumulation
Authors: Akiyama Tadakazu , Sugiyama Kazuyo , Shimizu Makiko , Tamaoki Tatsuya , Akinaga Shiro
Published: August 2005

7–Hydroxystaurosporine (UCN‐01), which was originally identified as a protein kinase C selective inhibitor, is currently in clinical trials as an anti‐cancer drug. We previously showed that UCN‐01 induced preferential G1–phase accumulation in tumor cells and this effect was associated with the retinoblastoma (Rb) protein and its regulatory factors, such as cyclin‐dependent kinase 2 (CDK2) and CDK inhibitors p21Cip1/WAF1and p27kipl. We demonstrate here that G1–phase accumulation was induced by UCN‐01 in Rb‐proficient cell lines (WiDr and HCT116 human colon carcinomas and WI‐38 human lung fibroblast), and it was accompanied by dephosphorylation of Rb. In addition, UCN‐01–induced G1–phase accumulation was also demonstrated in a Rb‐defective cell line (Saos‐2 human osteosarcoma), but not in a simian virus 40 (SV40)‐transformed cell line (WI‐38 VA13). Apoptosis was induced by UCN‐01 in the two Rb‐deficient cell lines, but not in the other Rb‐proficient cell lines. These observations suggest that G1–checkpoint function might be important for cell survival during UCN‐01 treatment. In addition, there may be a UCN‐01–responsive factor in the G1–checkpoint machinery other than Rb which is targeted by SV40. Further studies revealed a correlation between UCN‐01–induced G1–phase accumulation and reduction of cellular CDK2 kinase activity. This reduction was strictly dependent on down‐regulation of the Thr160–phosphor‐ylated form of CDK2 protein, and coincided in part with up‐regulation of p27Kip1, but it was independent of the level of the p21Cip1/WAF1protein. These results suggest that G1–checkpoint function, including a CDK2–regulatory pathway, may be a significant determinant of the sensitivity of tumor cells to UCN‐01.
Evaluation of Antivascular and Antimitotic Effects of Tubulin Binding Agents in Solid Tumor Therapy
Authors: Nihei Yukio , Suzuki Manabu , Okano Akira , Tsuji Takashi , Akiyama Yukio , Tsuruo Takashi , Saito Sachiko , Hori Katsuyoshi , Sato Yasufumi
Published: August 2005

Tubulin binding agents (TBAs) reduce tumor perfusion and inhibit mitosis of tumor cells in solid tumors, but it is not clear which effects contribute to the suppression of solid tumor growth. We evaluated the antivascular and antimitotic effects of several TBAs, combretastatin A‐4 (CS A‐4) phosphate, AC‐7700, a novel CS A‐4 derivative, colchicine, E7010, and vinblastine, on subcutaneous (s.c.) murine colon26 adenocarcinoma (c26). Tolerable doses of vinblastine and E7010 strongly inhibited tumor growth and induced mitotic arrest of tumor cells without affecting tumor perfusion. Colchicine had no effect on tumor growth and perfusion. When the injected dose was increased to the lethal range, however, these drugs markedly reduced tumor perfusion and caused necrosis of tumor tissue. Within the tolerable dose range, AC‐7700 both strongly suppressed tumor growth and reduced tumor perfusion, and CS A‐4 phosphate also exhibited a moderate antivascular effect. To evaluate the contribution of antivascular activity of TBAs to tumor growth suppression, excluding their direct cytotoxic effect on tumor cells, we established c26/acr, which is resistant to TBAs in vitro. Although E7010 showed a reduced suppressive effect on s.c. c26/acr tumor growth as compared with its effect on wild‐type c26, AC‐7700 remained potent against both cell lines. These results indicate that TBAs exert antivascular and antimitotic effects on solid tumors with marked differently effective dose ranges from agent to agent, and that the antivascular effect of TBAs inhibits solid tumor growth independently of the direct cytotoxic effect on tumor cells.
Urinary Bladder Lesions after the Chernobyl Accident: Immunohistochemical Assessment of p53, Proliferating Cell Nuclear Antigen, Cyclin D1 and p21WAF1/Cip1
Authors: Romanenko Alina , Lee Chyi Chia R. , Yamamoto Shinji , Hori Taka‐aki , Wanibuchi Hideki , Zaparin Wadim , Vinnichenko Wladimir , Vozianov Alexander , Fukushima Shoji
Published: August 2005

During the 11‐year period subsequent to the Chernobyl accident, the incidence of urinary bladder cancer in Ukraine has increased from 26.2 to 36.1 per 100,000 population. Cesium‐137 (137Cs) accounts for 80–90% of the incorporated radioactivity in this population, which has been exposed to long‐term, low‐dose ionizing radiation, and 80% of the more labile pool of cesium is excreted via the urine. The present study was performed to evaluate the histopathological features and the immunohistochemical status of p53, p21WAF1/Cip1, cyclin D1 and PCNA (proliferating cell nuclear antigen) in urinary bladder mucos a of 55 males (49‐92 years old) with benign prostatic hyperplasia who underwent surgery in Kiev, Ukraine, in 1995 and 1996. Group I (28 patients) inhabiting radiocontaminated areas of the country, group II (17 patients) from Kiev city with less radiocontamination and a control group III (10 patients) living in so‐called “clean” areas of Ukraine were compared. In groups I and II, an increase in multiple areas of moderate or severe dysplasia or carcinoma in situ was seen in 42 (93%) of 45 cases. In addi tion, two small transitional cell carcinomas were found in one patient in each of groups I and II. Nuclear accumulation of p53, PCNA, cyclin D1, and to a lesser extent p21WAF1/Cip1, was significantly increased in both groups I and II as compared with the control group III, indicating possible transformation events or enhancement of repair activities, that may precede the defect in the regulatory pathway itself, at least in the G1 phase of the cell cycle. Our results suggest that early malignant transformation is taking place in the bladder urothelium of people in the radiocontaminated areas of Ukraine and that this could possibly lead sometime in the future to an increased incidence of urinary bladder cancer.
Inhibition by β‐Carotene of Upper Respiratory Tumorigenesis in Hamsters Receiving Diethylnitrosamine Followed by Cigarette Smoke Exposure
Authors: Furukawa Fumio , Nishikawa Akiyoshi , Kasahara Ken‐ichiro , Lee In‐Seon , Wakabayashi Keiji , Takahashi Michihito , Hirose Masao
Published: August 2005

In recent intervention studies, β‐carotene failed to reduce or even increased the incidence of lung cancers in smokers. In the present investigation, the modifying effects of β‐Carotene at various doses on the development of upper respiratory tract tumors were investigated in Syrian hamsters treated with diethylnitrosamine (DEN) and cigarette smoke. A total of 120 male 5‐week‐old hamsters were divided into 4 groups, each consisting of 30 animals. After a sing le subcutaneous (s.c.) injection of 100 mg/kg DEN, hamsters in groups 1–4 were respectively administered diets supplemented with β‐carotene at doses of 0.5%, 0.05%, 0.005% or 0% during experimental weeks 1 to 13, and simultaneously exposed to cigarette smoke. The duration of cigarette smoke exposure was 9 min twice a day, 5 days a week. Because of a marked reduction of body weight in group 1, the highest dose of β‐carotene was changed to 0.25% after 10 days. In all groups, epithelial hyperplasias and/or papillomas were induced in the larynx and trachea. However, the incidence and multiplicity of papillomas in group 1 were significantly (P<0.05) lower than the group 4 values. Moreover, the β‐carotene treatments significantly (P<0.05 or 0.01) reduced both the incidence and multiplicity of hyperplasias in a dose‐dependent manner. The levels of retinol and β‐carotene in the serum, and the retinol level in the liver, were also elevated with dose dependence. Our results thus indicate that β‐carotene inhibits tumorigenesis, even at the high dose of 0.25%, under the present experimental conditions.