bluuebunny/crossref_metadata_2025 huggingface dataset:
Lysis of Fresh Human Tumor Cells by Autologous Peripheral Blood Lymphocytes and Tumor‐infiltrating Lymphocytes Activated hy PSK
Authors: Kariya Yoshitaka , Okamoto Norihiko , Fujimoto Takuya , Inoue Naoya , Kihara Takeshi , Sugie Katsuji , Yagita Masato , Kanzaki Hideharu , Mori Takahide , Uchida Atsushi
Published: August 2005

The protein‐bound polysaccharide PSK was tested for the ability to induce in vitro autologous tumor killing (ATK) activity in human cancer patients. Peripheral blood lymphocytes (PBL) and tumor‐infiltrating lymphocytes (TIL) demonstrated various levels of cytotoxicity against autologous, freshly isolated tumor cells. When PBL and TIL were cultured overnight with PSK, ATK activity was induced in previously non‐reactive cases and augmented in previously reactive samples. The PSK effect was observed with PSK concentrations of 10–100 μg/ml that could be obtained in the blood of cancer patients who received standard oral administration of PSK. The manifestation of PSK‐induced ATK required active cell metabolism and RNA and protein syntheses, but not DNA synthesis of lymphocytes. PSK‐induced enhancement of ATK was not abrogated by monoclonal antibodies (mAb) directed against interferon (IFN)α or IFNγ‐. In addition, mAb that neutralized interleukin‐2 (IL‐2) or mAb reactive with α‐chain or β‐chain of IL‐2 receptors (IL‐2R) had no effect on PSK‐induced ATK activity. Supernatants from PSK‐stimulated lymphocyte cultures did not induce ATK. Cell fractiona‐tion experiments revealed that CD3‐CD16+ large granular lymphocytes (LGL) and/or CD3+CD16‐ T lymphocytes were responsible for both spontaneous and PSK‐induced ATK. PSK‐activated LGL, but not T lymphocytes expressed lysis of fresh allogeneic tumor cells. These results indicate that PSK activates PBL and TIL to exhibit ATK independently of IL‐2/IL‐2R systems.
Inhibition of the Action of a Stimulatory GDP/GTP Exchange Protein for smg p21 hy Acidic Membrane Phospholipids
Authors: Kawamura Shiro , Kaibuchi Kozo , Hiroyoshi Motoki , Fujioka Hiroyuki , Mizuno Takakazu , Takai Yoshimi
Published: August 2005

A stimulatory GDP/GTP exchange protein for smg p21 (smg p21 GDS) stimulated the dissociation of GDP from smg p21B. This reaction was inhibited by acidic membrane phospholipids such as phosphatidylinositol, phosphatidylinositol‐4‐monophosphate, phosphatidylinositol‐4‐,5‐bisphosphate, phosphatidic acid, and phosphatidylserine but not by phosphatidykholine or phosphatidylethanolamine. These acidic phospholipids inhibited the smg p21 GDS action in a manner competitive with both smg p21 GDS and smg p21B. smg p21 GDS has other actions to inhibit the binding of smg p21B to membranes and to induce the dissociation of prebound smg p21B from the membranes. The acidic phospholipids also inhibited these two actions of smg p21 GDS. smgp21B has a polybasic region and an isoprenoid moiety in its C‐terminal region which are necessary for its membrane‐binding activity and its sensitivity to the smg p21 GDS actions. Therefore, it is possible that acidic membrane phospholipids interact with this polybasic region and thereby inhibit the smg p21 GDS actions.
Changes in Carboxylesterase Isoenzymes of Rat Liver Microsomes during Hepatocarcinogenesis
Authors: Maki Takako , Hosokawa Masakiyo , Satoh Tetsuo , Sato Kiyomi
Published: August 2005

Among the three major Carboxylesterase isoenzymes, RH1, RL1 and RL2, present in microsomes from normal rat liver, RL2 shows hydrolyzing activity towards 12‐O‐tetradecanoylphorboI‐13‐acetate and l‐oleoyl‐2‐acetyl‐rac‐glycerol, both activators of protein kinase C. Since protein kinase C has been suggested to be involved in carcinogencsis and cell proliferation, alterations in hepatic micro‐somal Carboxylesterase isoenzymes including RL2 were studied during hepatocarcinogenesis induced by the Solt‐Farber model. Alteration of RL2 was determined by measuring acetanilide‐hydrolyzing activity, by quantifying the protein amount using the single radial immunodiffusion method, and by activity staining following electrophoresis of liver microsomes. The isoenzyme composition of hepatic microsomal Carboxylesterase was changed after partial hepatectomy, and marked decreases in RL2 activity and protein content were observed at 4 weeks, at the time of preneoplastic foci induction. Partial hepatectomy alone also resulted in decreased RL2 activity. These findings suggest that RL2 may be involved in regulation of protein kinase C activity by metabolizing its activators at an early stage of hepatocarcinogenesis in rats.
Stimulatory Effects of Retinoic Acid on Tumor Growth and Serum Insulin‐like Growth Factor‐1 in Rats Bearing Estrogen‐responsive Pituitary Tumor MtT/Se
Authors: Roy Bidyut , Ogundigie Peter O. , Fujimoto Nariaki , Ito Akihiro
Published: August 2005

MtT/Se is one of 4 cell lines derived from an estrogen‐dependent pituitary tumor, MtT/F84. The main difference between these tumor types is that MtT/F84 secretes both growth hormone (GH) and prolactin (PRL) whereas MtT/Se secretes only GH. MtT/Se grew slowly in ovariectomized (ovex) rats, but tumor growth was much faster in estrogen‐treated ovex rats. Effects of dietary retinoic acid (RA) on tumor growth, serum GH and insulin‐like growth factor‐1 (IGF‐1) levels were examined in ovex rats. Latency of tumor growth was shortened, and tumor take and weight were promoted by all‐trans RA both in the presence and absence of exogenous estrogen. Serum GH and IGF‐1 levels became increased in tumor‐hearing rats whereas PRL levels remained unchanged. Serum IGF‐1 levels exhibited a good correlation with tumor weights (r=0.84). Our results suggest a close relationship between increase of tumor weight and stimulation of serum IGF‐1 level by RA in tumor‐bearing rats.
Relation between Nucleolar Size and Growth Characteristics in Small Cell Lung Cancer Cell Lines
Authors: Matsumoto Takeo , Terasaki Takeo , Mukai Kiyoshi , Wada Makio , Okamoto Aikou , Yokota Jun , Yamaguchi Ken , Kato Kanefusa , Nagatsu Toshiharu , Shimosato Yukio
Published: August 2005

Relationships among cytological features, doubling time, S‐phase percentage, expression of myc‐family oncogenes, DNA ploidy and biochemical properties were studied in thirteen small cell lung cancer cell lines. Six cell lines that grew slowly (average doubling time 99 h) and had lower S‐phase percentages (average 32%) showed inconspicuous nncleoli (average area of 1.5 μm2), and the remaining seven cell lines that grew quickly (average doubling time 45 h) and had higher S‐phase percentages (average 44%) showed large and prominent nncleoli (average area of 6.1 μm2). DNA index value obtained from flow cytometric DNA histograms showed that all cell lines except for H‐69 cell line displayed aneuploidy. Ribbon‐like cell arrangements were observed in the 7 cell lines that grew quickly, and in 1 cell line that grew slowly. Biochemically, six slow‐growing cell lines and four fast‐growing cell lines showed high levels of aromatic L‐amino acid decarboxylase activity, while in the remaining three fast‐growing cell lines its level was low. A high level of c‐myc or N‐myc oncogene expression was observed in all 7 cell lines that grew quickly, but not in any of the 6 cell lines that grew slowly. It appears that small cell lung cancer cell lines that grow quickly can be expected to have large nucleoli and ribbon‐like cell arrangements and to express high levels of mycfamily oncogenes, and that nucleolar size is a good indicator for growth characteristics.
Nuclear p53 Immunoreaction Associated with Poor Prognosis of Breast Cancer
Authors: Iwaya Keiichi , Tsuda Hitoshi , Hiraide Hoshio , Tamaki Kuniyoshi , Tamakuma Syouetsu , Fukutomi Takashi , Mukai Kiyoshi , Hirohashi Setsuo
Published: August 2005

p53 protein has been frequently detected at high levels in the nuclei of human breast cancer cells. We analyzed inununohistochemically the association between nuclear localization of p53 protein and clinical and histological parameters of breast cancer patients. Surgically resected tissues of 73 primary breast cancers were processed by acetone fixation and paraffin embedding and examined using an anti‐p53 monoclonal antibody, Pabl801. p53 immunoreactivity was detected in the nuclei of cancer cells in 17 cases (23%). The nuclear p53 immunoreaction was closely associated with overexpression of c‐erbB‐2 protein (P<0.05), high histologic grade (P<0.01), advanced clinical stage (P<0.05), and negative estrogen receptor status (P< 0.01). When 31 cases which had been followed up for more than 50 months were examined, a positive nuclear p53 immunoreaction was found to he significantly associated with shorter overall survival of patients (P<0.01). These results suggest that inununohistochemical examination of nuclear p53 protein is clinically useful as an indicator of breast cancer aggressiveness.
Detection of Epstein‐Barr Virus DNA in a Japanese Case of Lymphoepithelioma‐like Thymic Carcinoma
Authors: Matsuno Yoshihiro , Mukai Kiyoshi , Uhara Hisashi , Akao Ichiro , Furuya Shuichiroh , Sato Yuichi , Hirohashi Setsuo , Shimosato Yukio
Published: August 2005

Epstein‐Barr virus DNA was detected in a case of lymphoepithelioma‐like thymic carcinoma. A homogeneous terminal structure of the viral DNA was demonstrated in this case, indicating the presence of the viral genome in clonally expanded tumor cells. Since all of 26 other thymic epithelial tumors (eight non‐invasive, 13 invasive thymomas and four non‐lymphoepithelioma‐like thymic carcinomas) in Japanese were negative by polymerase chain reaction, it is suggested that lymphoepithelioma‐like thymic carcinoma may represent a unique pathological entity distinct from Epstein‐Barr virus‐negative thymic epithelial tumors, which are in the majority in Japan.
Production of Multiple Growth Factors by a Newly Established Human Thyroid Carcinoma Cell Line
Authors: Tohyama Kaoru , Yoshida Yataro , Ohashi Kensaku , Sano Emiko , Kobayashi Hisataka , Endo Keigo , Naruto Masanobu , Nakamura Toru
Published: August 2005

A multiple growth factor‐producing tumor cell line (NIM‐1) was newly established from a patient with thyroid cancer and remarkable neutrophilia. NIM‐1 cells also caused severe neutrophilia in nude mice bearing tumors. NIM‐1‐conditioned medium (NIM‐1CM) contained activities that supported not only granulocyte, macrophage and eosinophil colony formation of human bone marrow cells but also the growth of colony‐stimulating factor (CSF)‐dependent cell lines, NFS60‐KX and TF‐1. Northern blot hybridization analysis revealed the constitutive expression of granulocyte‐CSF (G‐CSF), granulocyte/macrophage‐CSF (GM‐CSF) and interleukin(IL)‐6 mRNAs in NIM‐1 cells. Enzyme‐linked immunosorbent assays (ELISA) using NIM‐1CM also confirmed the production of IL‐la and a small amount of IL‐1β besides G‐CSF, GM‐CSF and IL‐6 in NIM‐1 cells. In addition, unexpected production of IL‐11 in NIM‐1 cells was detected by northern blot hybridization analysis and by bioassay using an IL‐11‐dependent cell line. Therefore, NIM‐1 cell line is shown to produce multiple cytokines including potentially megakaryopoietic growth factors such as GM‐CSF, IL‐6 and IL‐11.
Somatic Mutation during Metastasis of a Mouse Fibrosarcoma Line Detected by DNA Fingerprint Analysis
Authors: Takada Toshinori , Suzuki Satoshi , Sugawara Yoshiko , Kominami Ryo , Arakawa Masaaki , Niwa Ohtsura , Yokoro Kenjiro
Published: August 2005

Metastatic nodules were examined by DNA fingerprint analysis. The probes used, Pc‐1 and Pc‐2, detect mutations as shifts in bands of the minisatellite loci which are dispersed among chromosomes. Four clonal lines of a fibrosarcoma from an F1 mouse (C57BL/Ka × C3H/He) were selected for various metastatic potentials upon inoculation into syngeneic mice. These four lines exhibited many extra bands resulting from recombination and/or DNA slippage, indicating accumulation of mutations during the successive passages in mice. One of the four, a 505 cell line which had been passaged extensively in vitro and consisted of a heterogenous population, was inoculated into thirteen syngeneic mice, and gave rise to six lung metastatic nodules in two mice. All the nodules showed band‐patterns distinct from one another, although nodules within a given mouse tended to show similar patterns. When a genetically tagged 505‐05‐01 clone was analyzed, three of nine metastatic nodules obtained also revealed new bands. These results strongly suggest that somatic mutations occur at a high frequency during metastasis, providing direct evidence of genetic instability of the tumor cells.
Analysis of Cytosol CA15‐3, Carcinoembryonic Antigen, Estrogen and Progesterone Receptors in Breast Cancer Tissues
Authors: Miya Toshimichi , Watanabe Toru , Adachi Isamu , Itabashi Masayuki , Fukutomi Takashi , Yamaguchi Ken , Abe Kaoru , Arakawa Masaaki
Published: August 2005

Cytosol levels of carcinoembryonic antigen (CEA), CA15‐3, estrogen receptor (ER) and progesterone receptor (PgR) of 194‐ primary breast cancer tissues were measured. ER and PgR determined by enzyme immunoassay methods correlated inversely with Page's grades of histological atypia and mitotic rate. Cytosol CA15‐3 correlated inversely with the grades of tubular and nuclear atypia and positively with the mitotic rate. CA15‐3 correlated positively with ER and PgR. Cytosol CEA showed no correlation with the pathological grade or hormone receptor status. These results indicate that hormone receptor‐positive breast cancers tend to have more differentiated morphology and slower growth rate than those without those receptors and that the cytosol CA15‐3 level may reflect some of the intrinsic malignant potency, particularly the growth rate, of breast cancer. Cytosol CA15‐3 as well as the hormone receptor status may have prognostic value for patients with breast cancer.
A Comparison between Argon‐dye and Excimer‐dye Laser for Photodynamic Effect in Transplanted Mouse Tumor
Authors: Okunaka Tetsuya , Kato Harubumi , Konaka Chimori , Sakai Harumasa , Kawabe Hirofumi , Aizawa Katsuo
Published: August 2005

Photodynamic therapy (PDT) utilizing a hematoporphyrin derivative (HpD) as a sensitizer has become a viable option for the local treatment of neoplastic disease. The argon‐dye laser system is commonly used as a light source in this treatment modality. The excimer‐dye laser, on the other hand, delivers high‐energy red light in a pulsatile fashion. In this investigation, we treated BALB/c mice bearing mouse kidney sarcoma cell tumors with PDT using HpD at the dose of 5 mg/kg body weight as a photosensitizer and either a standard argon‐dye laser or the pulsatile excimer‐dye laser as the light source. At equal light energy doses (50 J/cm2), necrotic changes at depths averaging 4 mm from the tumor surface were obtained with the argon‐dye laser (200 mW power output) while tumor necrosis at depths exceeding 15 mm from the tumor surface was obtained using the excimer‐dye laser (6 mJ/pulse, 5 Hz). To determine the best conditions for photoirradiation with the excimer‐dye laser, tumor‐bearing mice were treated with different total light doses (10, 30 and 50 J/cm2), dose rates (1, 3 and 6 mJ/cm2), and frequencies (5,15 and 50 Hz) of light exposure. Our results indicate that the optimal effects obtained with the excimer‐dye laser are related to the total light dose used and the dose rate, but not to the frequency of light exposure.
Enhanced Tumor Localization of Monoclonal Antibody by Treatment with Kininase II Inhibitor and Angiotensin II
Authors: Noguchi Akinori , Takahashi Toshio , Yamaguchi Toshiharu , Kitamura Kazuya , Noguchi Akira , Tsurumi Hiroshi , Takashina Ken‐ichiroh , Maeda Hiroshi
Published: August 2005

The effect of kininase II inhibitor, enalapril, on the delivery of monoclonal antibody A7 to the targeted tumor was investigated using athymic mice bearing human colon cancer, SVV1116. Enalapril alone, which enhances tumor vascular permeability through the kinin‐generating cascade, did not increase the uptake of 125 I‐labeled A7 (125I‐A7) in SW1116 due to the systemic hypotension induced by its inhibitory effect on angiotensin converting enzyme. However, with combined angiotensin II (AT‐II) and enalapril treatment, a 2‐fold increase in the accumulation of 125I‐A7 was seen when compared to A7 alone. This marked increase was presumably due to increased tumor vascular permeability induced by enalapril combined with the absence of hypotension due to the actions of AT‐II. This approach might be useful in radioimmunodetection and immunotargeting chemotherapy using monoclonal antibody.
Shock Wave and THP‐Adriamycin for Treatment of Rabbit's Bladder Cancer
Authors: Hoshi Senji , Orikasa Seiichi , Kuwahara Masa‐aki , Suzuki Ken‐ichi , Shirai Syuuichi , Yoshikawa Kazuyuki , Nose Masato
Published: August 2005

Focused high‐energy shock waves (6,000 to 10,000 shots) were targeted under ultrasound guidance onto implanted urinary bladder cancer in rabbits to elucidate its effect. Although only focal necrosis of the tumor was seen following 6,000 to 10,000 shots daily for 3 days or following chemotherapy (THP‐adriamycin) alone, almost total tumor necrosis was observed following a combined shock‐wave therapy for one day and THP‐adriamycin administration, demonstrating an additive and/or synergetic effect on rabbit urinary bladder cancer.
Strain Dependency of Cell‐type Specificity and Onset of Lymphoma Development in Eμ‐myc Transgenic Mice
Authors: Akagi Kiwamu , Miyazaki Jun‐ichi , Yamamura Ken‐ichi
Published: August 2005

c‐myc is a nuclear proto‐oncogene that, when activated, induces malignancies in a variety of tissues. Most murine plasmacytomas and human Burkitt's lymphomas have been shown to carry a chromosomal translocation involving c‐myc and immunoglobulin genes. To study genetic or epigenetic factors that affect mye‐induced lymphoid cell tumors, we previously introduced the Eμ‐mycΔ A gene lacking its own promoter and first exon into two inbred strains of mice, C57BL/6 and C3H/HeJ. We observed three characteristic features in our transgenic mice. First, T cell lymphoma predominated in the C3H background. Second, both pre‐B and B cell lymphoma developed at equal frequency in C57BL/6 transgenic mice. Third, the average age of onset is earlier than that reported by other investigators. To test whether these characteristics are due either to the lack of the promoter region and first exon of the c‐myc gene in the construct or to the genetic background of the mice, we introduced Eμ‐myc gene containing the complete c‐myc gene into fertilized eggs of C57BL/6 and C3H/HeJ mice. The cell‐type specificity, differentiation‐stage specificity and the average age at onset of lymphoma development were not affected by the transgene construct.
Successful Graft of HTLV‐I‐transformed Human T‐Cells (MT‐2) in Severe Combined Immunodeficiency Mice Treated with Anti‐asialo GM‐1 Antibody
Authors: Ishihara Shiro , Tachibana Nobuyoshi , Okayama Akihiko , Murai Koichi , Tsuda Kazunori , Mueller Nancy
Published: August 2005

To develop an experimental model of adult T‐cell leukemia/lymphoma in small animals, severe combined immunodeficiency (SCID) mice treated with anti‐asialo GM‐1 antibody were inoculated with MT‐2 cells, a cell line transformed by the human T‐cell leukemia virus (HTLV–I). Three mice injected with 4 × 107 cells subcutaneously or intramuscularly developed tumors at or near inoculation sites. Immunofluorescent antibody (IFA) staining for HTLV–I structural protein, p19, revealed the specific antigen in the cytoplasm of most cells from tumors and the DNA signals of HTLV–I proviral DNA were also positive in cellular DNA by polymerase chain reaction assay with HTLV–I tax gene primers, SK43/SK44. The MT–2 cells did not invade in mouse organs.
Growth Inhibition by Anchorage‐deficiency Is Associated with Increased Level but Reduced Phosphorylation of Mutant p53
Authors: Takahashi Kazuhide , Suzuki Katsuo , Uehara Yoshimasa , Ono Tetsuo
Published: August 2005

Human breast carcinoma MCF‐7 cells seeded on type I collagen‐coated dishes were provided with an anchor via the collagen receptor, integrin, and grew as actively as those in plastic tissue culture dishes. In contrast, cells seeded on a layer of soft agar became anchorage‐deficient and their growth was significantly inhibited, although the cell viability and the cell cycle distribution were unaffected. Immunoprecipitation analysis revealed that mutant p53 was phosphorylated at tyrosine in the anchorage‐provided cells. In contrast, the p53 in the anchorage‐deficient cells was present in 2‐fold greater amount, but was phosphorylated to a lesser extent. Addition of a potent protein‐tyrosine kinase inhibitor, herbimycin A, to the anchorage‐provided cells caused an elevated level of p53, and inhibitions of cell proliferation and p53 phosphorylation, without interfering with the cell adhesion to the substratum. These results demonstrated that the growth inhibition by anchorage‐deficiency or by herbimycin A is associated with an elevated p53 level and reduced p53 phosphorylation at tyrosine.
Soluble Interleukin‐6 Receptor Is Released from Receptor‐bearing Cell Linesin vitro
Authors: Nakajima Toshiharu , Yamamoto Shinji , Cheng Ming , Yasukawa Kiyoshi , Hirano Toshio , Kishimoto Tadamitsu , Tokunaga Tohru , Honda Mitsuo
Published: August 2005

Soluble interleukin‐6 receptor (sIL‐6R) was found to be spontaneously released from human myeloma cell line U266 cells into culture supernatant, and was quantitatively measured with a fluorescence sandwich enzyme‐linked immunosorbent assay employing antibodies specific to IL‐6R. The supernatant IL‐6R was generated only from IL‐6R‐positive cell lines; myeloma cell lines RPMI8226 and RPMI1788, and myelomonocytic cell lines U937, THP‐1, and HL‐60. In contrast, it was not released from the IL‐6R‐negative cells; T cell line MoIt‐4 and Burkitt lympboma cell line Raji. SDS‐PAGE analysis of the soluble IL‐6R from U266 cells suggested a molecular weight of approximately 50‐55 kDa, 25–30 kDa smaller than the mature cell surface receptor. These results suggest that the generation of soluble IL‐6R may be a maker of myeloma cells and myelomonocytic cells.
Autoradiographic Analysis of Radiolabeled Anti‐carcinoembryonic Antigen Monoclonal Antibody CEA102 in Colorectal Cancer Using Computed Radiography
Authors: Satoh Tsuyoshi , Watanabe Tadashi , Tadokoro Masanori , Sakamoto Junichi , Murayama Hiroki , Itoh Katsuki , Sakuma Sadayuki , Takagi Hiroshi
Published: August 2005

Anti‐carcinoembryonic antigen monoclonal antibody (MAb) CEA102 was produced by immunization with purified CEA and the specific accumulation of radiolabeled CEA102 in colorectal cancers was investigated by autoradiography of surgical specimens using Fuji Computed Radiography (FCR). Five patients with colorectal cancer were injected intravenously with 131I‐labeled intact CEA102 or its F(ab′)2. Primary tumor and liver metastases were successfully detected by external scanning with a gamma camera in 4 cases. Autoradiographic study of the surgical specimens using FCR showed predominant localization of 131I‐labeled CEA102 in primary tumors and liver metastases in all cases. Even a small liver metastasis (0.5 cm) was clearly visualized in the autoradiogram by FCR. The pixel distribution curves of the density of the respective tissues in the autoradiograms by FCR showed the heterogeneity of the distribution of administered radiolabeled MAb in individual tumors, but the density of the tumors was higher than that of the normal tissues. In the quantitative distribution analysis of CEA102, the uptake of the primary tumor (mean 1.10%ID/kg) was ten‐fold greater than that of the normal colon mucosa (mean G.10%ID/kg). These results revealed that the application of MAb has great potential in radioimmunodetection as well as in antibody‐directed therapy.
Metabolism of Tegafur in Rat Liver Observed by in vivo 19F Magnetic Resonance Spectroscopy and Chromatography
Authors: Harada Masafumi , Nishitani Hiromu , Koga Keiko , Miura Iwao , Umeno Yukihiko
Published: August 2005

Metabolism of tegafur in the rat liver was observed by in‐vivo19F magnetic resonance spectroscopy (MRS). After MRS observation, tegafur and q‐fluorouracil (S‐FU) in the liver were determined by a shromatographic method for comparison with the results of 19F‐MRS. Rats were divided into 3 groups: 1) CCl4‐induced liver injury group, 2) uracil combined group, 3) control group. Catabolism to fluoro‐β‐alanine was suppressed in both the liver injury group and the uracil combined group. Low peaks of 5‐FU and fluoronucleotides could be found only in the uracil combined group. The result of 19F MRS observation of each group was in agreement with the result of determination of tegafur and 5‐FU by chromatography. This showed that substances which could be observed by 19F‐MRS were in proportion to all intracelluar fluoro‐containing substances. 19F‐MRS can provide direct information on the metabolism of fluoropyimidines non‐invasivey and it might be a useful aid in choosing suitable shemotherapy for patients.
Effects of Antitumor Agents on Subcutaneous Implants and Hepatic Metastases of Colon Carcinoma 26 in Mice
Authors: Iigo Masaaki , Nishikata Ken‐ichi , Nakajima Yoko , Araki Eiji
Published: August 2005

We investigated the responses of experimentally produced hepatic metastases of colon carcinoma 26 tumor and subcutaneously (SC) implanted colon carcinoma 26 tumor in mice to 17 clinically used and one under‐development antitumor agents using same dose regimen. In intravenous administrations on days 7 and 14, there were no significant differences in their responses to most of the tested agents. However, there were big differences in the responses to some of the agents. Nimustine more effectively prolonged the lifespan of SC implanted tumor‐bearing mice than of mice bearing hepatic metastases. Mitomycin C was, however, considerably more effective on hepatic metastases than on SC implanted tumor. ME2303, a new fluorinated anthracycline derivative, showed a similar effect to doxorubicin on both tumors. However, administrations of ME2303 on days 7,11 and 15 showed more marked antitumor effect only on hepatic metastases than administrations on days 7 and 14. Doxorubicin was less active against both tumors for administrations on days 7, 11 and 15 than for those on days 7 and 14. These results suggest the importance of the site of tumor growth for the action of some drugs. ME2303 may be active against hepatic metastases if it is administered by multiple injections.
Cell‐killing Activity and Kinetic Analysis of a Novel Antitumor Compound NC‐190, a Benzo[a]phenazine Derivative
Authors: Nakaike Shiro , Yamagishi Takehiro , Nanaumi Kayoko , Otomo Susumu , Tsukagoshi Shigeru
Published: August 2005

A novel antitumor compound, N‐β‐dimethylaminoethyl 9‐carboxy‐5‐hydroxy‐10‐methoxybenzo[a]‐phenazine‐6‐carboxamide sodium salt (NC‐190), was evaluated for antitumor activity in vitro against cultured tumor cell lines, and the kinetics of cell killing was elucidated. NC‐190 strongly inhibited the growth of all of 3 murine tumor cell lines, 7 human tumor cell lines and 2 normal cell lines. With continuous exposure, the 50% inhibition concentrations were in the range of 0.005–0.06 μg/ml, except for KATO‐III (2.15 μ g/ml). By colony‐forming assay, concentrations of NC‐190 giving 90% cell kill (IC90) at various exposure times were obtained with HeLa S3 cells. The plot of IC90exposure time on a log‐log scale was linear for NC‐190 with a slope of ‐1, which is typical for cell cycle phase‐nonspecific agents. A 2 h treatment with NC‐190 induced a rapid reduction in cell viability at doses of more than 3 μ g/ml. At the dose where colony formation was completely inhibited, cell viability was persistently reduced to below 20% during the cell culture period. NC‐190 cauced a dose‐ and time‐dependent reduction in DNA synthesis. The inhibitions of RNA and protein synthesis were less than that of DNA synthesis. Spectroscopic studies of NC‐190 mixed with calf thymus DNA demonstrated that NC‐190 was capable of interacting with DNA. However, DNA thermal denaturation studies suggested that intercalation of NC‐190 was weak in comparison with those of classical intercalating drugs.
Low Frequency of Rearrangements of the ret and trk Proto‐oncogenes in Japanese Thyroid Papillary Carcinomas
Authors: Wajjwalku Worawidh , Nakamura Shigeo , Hasegawa Yasuhisa , Miyazaki Koichi , Satoh Yasuyuki , Funahashi Hiroomi , Matsuyama Mutsushi , Takahashi Masahide
Published: August 2005

We investigated the frequency of rearrangements of the ret and trk proto‐oncogenes in Japanese thyroid tumors. DNAs from 38 thyroid papillary carcinomas and 14 follicular adenomas were analyzed by Southern blotting. Rearrangements of the ret and trk proto‐oncogenes were detected in one and two papillary carcinomas, respectively, but not in follicular adenomas. Analysis by a reverse transcriptase‐polymerase chain reaction method showed that the ret rearrangement‐positive tumor contained the PTC/refTPCchimeric transcript, which was reported to be found specifically in thyroid tumors and adenomatous goiter. We also found that rearranged mRNA of the trk proto‐oncogene was expressed at high levels in one of two trk rearrangement‐positive tumors. Our results indicated that the frequency of rearrangements of these proto‐oncogenes in Japanese papillary carcinomas was much lower than that in Italian patients.
CD7 Expression in Malignant Pleural Mesothelioma
Authors: Hibi Shigeyoshi , Kataoka Yoshiko , Fujita Naohisa , Okabe Hidetoshi , Yoshikawa Kazuhiro , Seto Masao , Ueda Ryuzo , Imashuku Shinsaku
Published: August 2005

CD7 antigen was found to be expressed on malignant mesothelioma arising from the right pleura in a 15‐year‐old girl not only by immunostaining using monoclonal antibodies, but also by Northern blot analysis. The level of expression in this tumor was comparable to those in T‐cell lines, Jurkat and CCRF‐CEM. Cytogenetic analysis of the tumor showed hypodiploidy (n = 43). CD7 has been regarded as one of the hematopoietic cell markers selectively expressed on the majority of T cells and multipotential stem cells. To our knowledge, this is the first report of a non‐hematopoietic tumor expressing CD7.
Potentiation by Tumor Necrosis Factor of Mitoxantrone Cytotoxicity to Human Ovarian Cancer Cell Lines
Authors: Russo Patrizia , Parodi Silvio , Billi Giovanna , Oliva Cristina , Venturing Marco , Noviello Elvira , Conte PierFranco
Published: August 2005

The cytotoxic activity of human recombinant tumor necrosis factor (rHuTNF) (from 0.01 to 10000 U/ml) was assayed on six human ovarian cancer cell lines and one human cervical carcinoma cell line using a crystal violet assay. rHuTNF was cytotoxic to four cell lines (A2780, A2774, SW626, PAD, while 3 cell lines (IGROV1, SKOV3, Mel80) were marginally sensitive to its activity. However, under the same experimental conditions rHuTNF markedly enhanced the cytotoxicity of mitoxantrone, a chemotherapeutic drug targeted at DNA topoisomerase II, in six cell lines. The potentiation of mitoxantrone cytotoxicity was not caused by increased drug accumulation after rHuTNF treatment. No significant increase in cytotoxicity to Me180 cell line was seen when rHuTNF was added to mitoxantrone.
Promoting Effects of Potassium Dibasic Phosphate on Early‐stage Renal Carcinogenesis in Unilaterally Nephrectomized Rats Treated with N‐Ethyl‐N‐hydroxyethylnitrosamine
Authors: Hiasa Yoshio , Konishi Noboru , Nakaoka Shingo , Nakamura Toshimitsu , Nishii Kiyoji , Ohshima Masato
Published: August 2005

The effects of potassium dibasic phosphate (PDP), potassium aluminum sulfate (PAS) and copper sulfate (CS) on early‐stage renal carcinogenesis were investigated in unilaterally nephrectomized male Wistar rats after N‐ethyl‐N‐hydroxyethylnitrosamine (EHEN) administration. After feeding 1,000 ppm EHEN, or basal diet for 2 weeks and removal of the left kidney at week 3, male Wistar rats were divided into 8 groups of 20 rats each. These groups received the following dietary treatments: 50,000 ppm PDP, 50,000 ppm PAS, 5,000 ppm CS or basal diet, respectively, for 18 weeks from weeks 3 to 20. The average numbers of adenomatous hyperplasias counted as preneoplastic lesions in the EHEN with 50,000 ppm PDP group were significantly higher than in the EHEN alone group or the EHEN followed by 50,000 ppm PAS or 5,000 ppm CS group. The treatment with 50,000 ppm PDP induced renal calcification and promoted the development of preneoplastic lesions in unilaterally nephrectomized rats treated with EHEN, but that with 50,000 ppm PAS or 5,000 ppm CS did not.
Efficacy and Specificity of a Monoclonal Antibody‐Drug Conjugate in Chemotherapy by Intratumoral Injection
Authors: Kitamura Kazuya , Takahashi Toshio , Miyagaki Takuya , Yamaoka Nobuki , Tsurumi Hiroshi , Ohtsuji Eigo , Kamiguchi Michiko , Noguchi Akinori , Yamaguchi Toshiharu
Published: August 2005

The murine monoclonal antibody (Mab) A7 conjugated to neocarzinostatin (A7‐NCS) was injected intratumorally (IT) into tumor bearing nude mice. Its pharmacokinetics and tumoricidal effects were compared in the high, moderate and low antigen expressing xenograft for SW1116, WiDr and KB tumor‐bearing nude mice, respectively. When injected IT into nude mice, [125I]A7‐NCS was retained in the tumors according to the degree of antigen expression; it was also disseminated into the blood inverse proportion to the antigen expression. Addition of an excess amount of Mab A7 reduced [125I]‐A7‐NCS accumulation in SW1116 xenograft and elevated the [125I]A7‐NCS concentration in the circulation. Complete tumor reduction was found in all 5 mice with SW1116 tumor, and 2 of 5 mice with WiDr tumor. However, only incomplete tumor suppression was observed in mice with the KB tumor. The significant tumor reduction in SW1116 bearing nude mice was attenuated when excess of Mab A7 was simultaneously administered with A7‐NCS‐ These findings indicate that A7‐NCS was localized in the target tumors and exerted its tumoricidal effects depending on the degree of antigen‐antibody interaction when administered IT. Thus, A7‐NCS can be used successfully in vivo for local therapy, auguring new and promising applications for local cancer therapy.
Antitumor Effect of PSK at a Distant Site: Tumor‐specific Immunity and Combination with Other Chemotherapeutic Agents
Authors: Ebina Takusaburo , Murata Kazuko
Published: August 2005

The antitumor effect of PSK, a Coriolus preparation, was analyzed with the double grafted tumor system in which BALB/c mice received intradermal inoculations of syngeneic Meth‐A fibrosarcoma in the right (primary tumor, 105 cells) and left (distant tumor, 2 × 105 cells) flanks. Intratumoral administration of PSK significantly inhibited the growth of not only the right but also the left tumor. PSK also inhibited the growth of a methylcholanthrene‐induced fibrosarcoma BAMC‐1, and a methylurethane‐induced adenocarcinoma Colon 26 in the double grafted tumor system of syngeneic BALB/c mice. However, when the left distant tumor was different from the right Meth‐A tumor, the intratumoral administration of PSK in the right tumor was unable to inhibit the growth of the left’ BAMC‐1 or RL♂ ‐1 tumor. The PSK‐induced immunity, therefore, is tumor‐specific and T lymphocytes may play an important role in antitumor memory function. The enhancement of concomitant immunity by PSK treatment was completely impaired by previous intravenous administration of an alkylating agent, cyclophosphamide (CY). The enhancement of sinecomitant immunity by PSK treatment was also impaired by previous CY intravenous administration. The antitumor effect of PSK was suppressed by previous intravenous administration of another alkylating agent, ACNU. It is possible that alkylating agents suppress the function of effector T cells and granulocytes which are very important for the antitumor immune cascade reaction due to PSK treatment. On the other hand, the antitumor effect of PSK was enhanced by previous intravenous administration of an anti‐metabolite, 5‐fluorouraeil.
Simultaneous Expression of T‐Cell and Myeloid Cell Phenotypes in Eight Newly Established HTLV‐I‐positive T‐Cell Lines
Authors: Koizumi Shigeki , Sugiura Makoto , Zhang Xian‐Kui , Yamashiro Katsushige , Iwanaga Michiyo , Imai Shosuke , Osato Toyoro
Published: August 2005

Eight cell lines were established from patients with adult T‐cell leukemia, and from normal adults, by cocultivation with human T‐cell leukemia virus type I(HTLV‐I)‐producer cell lines in the presence of interleukin‐2. All of these cell lines harbored HTLV‐I and showed T‐cell markers CD2, CD3 and CD4, hut not B‐cell markers. Unexpectedly, all eight cell lines expressed a myeloid marker CD13 and three of the eight lines also expressed another myeloid marker CD33. Dual staining showed the simultaneous expression of CD3 and CD13 on the same cells. Thus, evidence was obtained for the expression of myeloid antigens on HTLV‐I‐harboring T cells.
Tumor Necrosis Factor‐a Stimulates Colony Formation by a Megakaryoblastic Leukemia Cell Line, CMK
Authors: Akiyama Yasuto , Yamaguchi Ken , Sato Takeyuki , Abe Kaoru
Published: August 2005

The CMK cell line is an acute megakaryoblastic leukemia cell line established from a patient with Down's syndrome, and is known to possess characteristics of normal megakaryocytes. Several cytokines with the ability to stimulate megakaryopoiesis, such as interleukin‐3 (IL‐3), interleukin‐6 (IL‐6) and granulocyte‐macrophage colony‐stimulating factor (GM‐CSF), stimulated colony formation by CMK cells. The present study revealed that tumor necrosis factor‐α (TNF‐α) stimulated colony formation by CMK cells; the potency was almost equal to that of IL‐3, IL‐6 or GM‐CSF. Scatchard plot analysis revealed that CMK cells possess two types of specific binding sites for TNF‐α. The high‐affinity binding sites had an affinity constant of 0.18 nM, and numbered 5,000. The low‐affinity binding sites had an affinity constant of 1.8 nM/and numbered 19,000. These results raise the possibility that TNF‐α can act as a growth‐stimulating agent on megakaryocyte‐Iineage cell line.
Menogaril, an Anthracycline Derivative, Inhibits DNA Topoisomerase II by Stabilizing Cleavable Complexes
Authors: Ono Katsuhiro , Ikegami Yoji , Nishizawa Miwako , Andoh Toshiwo
Published: August 2005

Menogaril, an anthracycline derivative, has been shown to possess antitumor activity in experimental animal systems, and is now under phase II clinical studies. However, its mechanism of action has not been elucidated. We have found that it inhibits the decatenation activity of purified DNA topoisomerase II using kinetoplast DNA from Crithidia fasciculata, its IC50 being 10 μM, which is comparable to that of etoposide. It does not, however, inhibit topoisomerase I activity at concentrations of up to 400 μM. Binding of topoisomerase II with DNA is not affected, but cleavable complex formation is stimulated by the drug. Cleavage site specificity differes from that of 4′‐(9‐acridinylamino)methanesulfon‐m‐anisidide. Menogaril was shown to possess a weak double‐helix unwinding activity. These findings allow us to classify menogaril as a cleavable complex‐stabilizing topoisomerase II inhibitor.
Atrophic Gastritis and Stomach Cancer Risk: Cross‐sectional Analyses
Authors: Kato Ikuko , Tominaga Suketami , Ito Yoshiaki , Kobayashi Seibi , Yoshii Yuri , Matsuura Akira , Kameya Akira , Kano Tomoyuki
Published: August 2005

The relationship between atrophic gastritis and stomach cancer risk was investigated in case‐control analyses involving 387 cases with stomach cancer and 5,422 control subjects who received gastroscopic examination at Aichi Cancer Center Hospital from April, 1985 to March, 1989. The presence of atrophic gastritis, the degree and extension of the atrophy and the presence of granularity and erosion were diagnosed endoscopically by six gastroenterologists. The prevalence of atrophic gastritis increased with age and was higher in males than in females. The relative risk (RR) of stomach cancer was S.13 (95% confidence interval (CI): 2.79–9.42) if a subject had any type of atrophic gastritis. The risk further increased with advancing degree of atrophy and increasing extension on the greater and lesser curvatures. The RR associated with severe atrophy was 7.73 (95% CI: 3.95–15.12). These associations remained significant when analyzed by sex and age. The presence of granularity and erosion did not much affect the estimated risks. A clear difference in risk appeared in the analyses by histological type of cancer. The RR associated with atrophic gastritis was 24.71 (95% CI: 3.46–176.68) for the intestinal type and 3.49 (95% CI: 1.77–6.87) for the diffuse type. These findings may suggest a need for intensive follow‐up of patients with severe atrophic gastritis.
DT‐5461, a New Synthetic Lipid A Analogue, Inhibits Lung and Liver Metastasis of Tumor in Mice
Authors: Sato Katsuaki , Saiki Ikuo , Yoo Yung Choon , Igarashi Yu , Kiso Makoto , Hasegawa Akira , Azuma Ichiro
Published: August 2005

We have investigated the antimetastatic effect of a new synthetic lipid A analogue, of low endo‐toxicity, DT‐5461, against two highly metastatic tumor cell lines, L5178Y‐ML25 T‐lymphoma and B16‐BL6 melanoma cells in mice. Four intermittent i.v. administrations of DT‐5461 at intervals of 4 days resulted in a significant inhibition of liver metastasis caused by i.v. injection of L5178Y‐ML25 cells and lung metastasis of B16‐BL6 cells in the experimental metastasis models. Intraperitoneal and intranasal administrations as well as i.v. administration of DT‐5461 were also effective in preventing lung metastasis of the melanoma cells. Multiple administrations of DT‐5461 before the surgical excision of primary tumors significantly reduced the number of lung colonies of melanoma cells and primary tumor size. Similarly, this treatment modality after the surgical excision of primary tumors showed a greater reduction of lung tumor colonies as compared with lipopolysaccharide, a synthetic lipid A (No. 506) and its analogue as well as untreated control in the spontaneous lung metastasis model. Furthermore, the group that received DT‐5461 after the inoculation of lymphoma or melanoma cells showed significantly enhanced survival rate compared with the untreated control. These results suggested that DT‐5461 may he therapeutically useful for the inhibition of tumor metastasis.
Inhibition by Fibrin Coagulation of Lung Cancer Cell Destruction by Human Interleukin‐2‐activated Killer Cells
Authors: Atagi Shinji , Sone Saburo , Fukuta Katsuyuki , Ogura Takeshi
Published: August 2005

We examined the effect of fibrin coagulation on tumor cytotoxicity mediated by human lymphokine (IL‐2)‐activated killer (LAK) cells. LAK cells were induced from peripheral blood mononuclear cells (MNC) by culture with recombinant IL‐2 for 4 or 5 days, and LAK cell‐mediated cytotoxicity against tumor cells was assessed by 51Cr release assay in the presence or absence of plasma from normal subjects and lung cancer patients. Plasma did not affect the phase of induction of LAK activity by IL‐2, but dose‐dependently inhibited the effector phase of LAK cell‐mediated cytotoxicity against Daudi cells. Similar inhibition of LAK cell‐mediated cytotoxicity was observed on pretreatment of Daudi cells and human lung cancer cell lines with human fibrinogen plus thrombin. A parallel relationship was found between the amount of fibrinogen in plasma of lung cancer patients and inhibition of LAK cytotoxicity. This inhibition was reduced by addition of anticoagulants (heparin or argatroban). These findings suggest that fibrin coagulation on tumor cells protects them from LAK cell‐mediated tumor cytotoxicity in malignant lesions and that a combination of an anticoagulant drug and IL‐2/LAK therapy may be effective for treatment of lung cancer patients.
Fluorouracil Catabolism in the Combination Treatment of Cyclophosphamide, Methotrexate and Fluorouracil
Authors: De Bruijn Ernst A. , Van Der Heyden Synke A. M. , Gheuens Eric E. O. , Maes Robert A. A.
Published: August 2005

The CMF‐regimen is amongst the most effective chemotherapeutic approaches in the treatment of breast cancer. It is generally accepted that the efficacy of the combination of the three agents used in the regimen, i.e., cyclophosphamide (CY), methotrexate (MTX) and fluorouracil (FUra), is based on interactions between the drugs at the intratumoral level. In WAG/Rij rats we previously demonstrated that change of FUra clearance at the first day of the CMF‐regimen occurs owing to concomitant CY+MTX. In the present study clearance of FUra and the first product of FUra catabolism, FUraH2, were monitored at day 1 and day 8 of the regimen upon treatment with single agent FUra (F), MTX + FUra (MF), CY + FUra (CF), and CY + MTX + FUra (CMF). At the first day of treatment, FUra and FUraH2 systemic exposure was demonstrated to be increased in CMF‐treated rats owing to concomitant CY + MTX. At the eighth day of treatment it was found that repeated CY administration during the previous seven days in CF‐treated rats resulted in increased FUra and FUraH2 systemic exposure and therefore increased the dose of FUra artificially. It is concluded that altered FUra clearance owing to extratumoral interactions by concomitant CY and MTX contributes to the efficacy of the CMF‐regimen.
Telomere Elongation Frequently Observed during Tumor Metastasis
Authors: Takada Toshinori , Hayashi Tatsuhiko , Arakawa Masaaki , Kominami Ryo
Published: August 2005

Changes in the number of telomere repeat arrays have been examined during metastasis of two mouse tumor cell lines. Telomeres were detected as bands and a smear by pulsed‐field gel electrophoresis and gel‐hybridization using (TTAGGG)4 as a probe. Very long size variants of telomeres were frequently observed in metastatic nodules. This suggests that at least some of the tumor cells have an ability to elongate telomeres. This elongation may compensate for the continuous loss of telomere repeats due to cell divisions, which would eventually lead to cell death.
Oligonucleotide Sequences Required for Natural Killer Cell Activation
Authors: Kuramoto Etsuro , Yano Osamu , Kimura Yoshimitsu , Baba Makoto , Makino Tadashi , Yamamoto Saburo , Yamamoto Toshiko , Kataoka Tetsuro , Tokunaga Tohru
Published: August 2005

Based on the previous finding that certain 30‐mer single‐stranded oligodeoxyribonucleotides (oligonucleotides) having particular 6‐mer palindromic sequences could induce interferon‐alpha and ‐gamma, and enhance natural killer activity, the present study was carried out to clarify the entire relationship between the activity and the sequence of 30‐mer oligonucleotides. The results indicated that the activity depended critically on the presence of particular palindromic sequences including the 5 ‐CG‐3 motif(s). The size and the number of palindromes as well as the extra‐palindromic sequences also influenced the activity. An oligonucleotide with a 10‐mer palindrome and extra‐palindromic oligoguanylate sequences showed the strongest activity among the oligonucleotides tested.
An Intrinsic Thymic Epithelial Abnormality Is Responsible for the Spontaneous Development of Predominantly Lymphocytic Thymomas in BUF/Mna Rats
Authors: Taguchi Osamu , Kontani Keiichi , Ikeda Hiroshi , Matsuyama Mutsushi
Published: August 2005

The nature of tumorigenesis of predominantly lymphocytic thymoma was examined using an animal model. Rats of the inbred BUF/Mna strain were found spontaneously to develop predominantly lymphocytic thymomas, histologically indistinguishable from their human counterparts, at an incidence of virtually 100%. Thymic rudiments of BUF/Mna rats grafted 17 months previously under the renal capsule of young athymic ACI/NMs‐rnu/rnu rats also gave rise to similar lesions. The lymphocytes in the thymomas expressed T‐cell antigens (rat Lyt‐1 and Lyt‐2.3), as in the normal case, and ACI rat specific antigen. When BUF/Mna rats of thymoma age were irradiated with a lethal dose of 12 Gy and then received a single injection of bone marrow cells (8 × 107) from BALB/c‐nu/nu mice, thymomas were re‐formed three weeks later (in 2 of 5 rats) with the replacement lymphocytes expressing mouse Thy‐1.2 antigen. These results indicate that an intrinsic thymic epithelial abnormality is responsible for the development of predominantly lymphocytic thymomas in BUF/Mna rats.