bluuebunny/crossref_metadata_2025 huggingface dataset:
Spermidine/Spermine N‐Acetyltransferase, a New Biochemical Marker for Epithelial Proliferation in Rat Bladder
Authors: Matsui‐Yuasa Isao , Otani Shuzo , Yano Yoshihisa , Takada Nobuyasu , Shibata Masa‐Aki , Fukushima Shoji
Published: August 2005

We examined the activity of spermidine/spermine N1‐acetyltransferase (SAT), a rate‐limiting enzyme of the biodegradation of polyamines, in N‐butyl‐N‐(4–hydroxybutyI)nitrosamine‐induced transitional cell carcinoma (TCC) and melamine‐induced papillomatosis of rat bladder, and compared the activity to that of ornithine decarboxylase (ODC). Both activities were higher in both lesions than in control rats. The difference between SAT and ODC activities in cancerous tissue and papillomatosis was not significant. Cells stained for proliferating cell nuclear antigen (PCNA) were abundant in papillomatosis. TCC had areas with much PCNA. The results indicated that an elevation of SAT activity occurs in both reversible and irreversible proliferation of bladder epithelium and could be important in bladder carcinogenesis.
Species Difference among Experimental Rodents in Induction of P450IA Family Enzymes by 2‐Amino‐1‐methyl‐6‐phenylimidazo[4,5‐b]pyridine
Authors: Degawa Masakuni , Kobayashi Ken‐ichiro , Miura Shin‐ichi , Arai Hiroshi , Esumi Hiroyasu , Sugimura Takashi , Hashimoto Yoshiyuki
Published: August 2005

Rats, mice, hamsters and guinea pigs were given an i.p. injection of 2‐amino‐1‐methyl‐6‐phenyl‐imidazo[4,5‐b]pyridine (PhIP), a protein‐derived pyrolysate component present in cooked foods, and inductions of cytochrome P450 (P450) in the liver and kidney of these animals were examined. The activity and amount of P450s corresponding to the rat P4S0IA1 and P450IA2 were assessed by means of a bacterial mutation test using 3 carcinogenic heterocyclic aromatic amines including PhIP as substrates and by Western blotting with a monoclonal antibody reactive with both P4S0IA1 and P450IA2. In rats, PhIP induced P450IA1, P450IA2 and a new but unspecified P450 isozyme in the liver, and induced P450IA1 in the kidney. However, PhIP induced none of these P450 isozymes in mice, hamsters and guinea pigs.
Inactivation of the p53 Gene Is not Required for Tumorigenesis of Medullary Thyroid Carcinoma or Pheochromocytoma
Authors: Yana Ikuo , Nakamura Tsutomu , Shin Eisei , Karakawa Katsu , Kurahashi Hiroki , Kurita Yoshihiro , Kobayashi Tetsuro , Mori Takesada , Nishisho Isamu , Takai Shin‐ichiro
Published: August 2005

A polymerase chain reaction (PCR)‐mediated RNase protection analysis was performed to detect subtle genetic alterations of p53 in medullary thyroid carcinoma (MTC) and pheochromocytoma. None of the 30 pheochromocytomas showed abnormal RNase protection patterns. Only one of 32 MTCs showed an abnormal pattern, and subsequent DNA sequencing of the PCR product revealed that it had a G to C transversion in codon 49 that resulted in a change from aspartic acid to histidine. However, this was a sporadic MTC with no specific clinicopathological characteristics. On the basis of a previous report that genes on chromosome 17p were not deleted in MTCs and were relatively infrequently deleted in pheochromocytomas, our results suggest that the p53 gene is not involved in tumorigenesis of MTC or pheochromocytoma.
Effect of Pravastatin, a Potent 3‐Hydroxy‐3‐methylgliitaryl‐coenzyme A Reductase Inhibitor, on Survival of AH130 Hepatoma‐bearing Rats
Authors: Kawata Sumio , Kakimoto Hiroki , Ishiguro Hiroshi , Yamasaki Eiji , Inui Yoshiaki , Matsuzawa Yuji
Published: August 2005

3‐Hydroxy‐3‐methylglutaryl‐coenzyme A (HMG‐CoA) reductase inhibitor is known to have an inhibitory effect on cell growth in addition to a cholesterol‐lowering effect. This study examined the effect of pravastatin, a potent inhibitor of HMG‐CoA reductase, on the survival of AH130 hepatoma‐bearing rats. Pravastatin (1, 2, or 8 mg/kg body weight) was intraperitoneally injected once a day into tumor‐bearing rats. The difference in the survival curves was significant between the controls and the rats treated with S mg/kg of pravastatin (P < 0.019 by logrank test) but not between the controls and the rats treated with 1 or 2 mg/kg of the inhibitor. The tumor volume was significantly decreased in the rats treated with 8 mg/kg of pravastatin (P < 0.05). These observations showed that intraperitoneal injection of pravastatin could improve the survival of AH130 hepatoma‐bearing rats and had an inhibitory effect on the growth of the ascites form tumor.
Accelerated Appearance of Skin Tumors in Hairless Mice by Repeated UV Irradiation with Initial Intense Exposure and Characterization of the Tumors
Authors: Nishigori Chikako , Tanaka Miran , Moriwaki Shin‐ichi , Imamura Sadao , Takebe Hiraku
Published: August 2005

Skin tumors were produced on the back of hairless mice, HOS (HR/De), by exposure to ultraviolet B light (UVB, 290–320 nm) with 4 different protocols. The first tumors appeared earlier (in 10 weeks in group I and 7 weeks in group III) when initial intense exposure was given, followed by repeated lower‐level exposures, than when the mice were exposed to the repeated UV only (in 16 weeks both in group II and group IV). All mice developed skin tumors earlier in the groups given the repeated UV exposures three times a week than in the groups given the exposures twice a week. Most of the skin tumors produced by the UVB exposure were histologically malignant, being transplantable to nude mice, and the cultured cells grown from the tumors were capable of producing tumors when injected into nude mice. The accelerated development of skin tumors by initial intense exposure and short intervals of repeated exposure observed in this study may have implications for humans who expose themselves to intense sunbathing and UV tanning (burning) by fluorescent sun lamps.
Evidence of Direct Generation of Oxygen Free Radicals from Heterocyclic Amines by NADPH/Cytochrome P‐450 Reductase in vitro
Authors: Sato Keizo , Akaike Takaaki , Kojima Yuichiro , Ando Masayuki , Nagao Minako , Maeda Hiroshi
Published: August 2005

Highly purified cytochrome P‐450 reductase (also called cytochrome c reductase; EC 1.6.2.4.) and NADPH were used to generate superoxide radical (O2) from 11 different heterocyclic amines (HCAs) as identified by electron spin resonance spectroscopy using the spin trapping method with 5,5‐dimethyl‐1‐pyrroline JV‐oxide (DMPO). The signal intensity of DMP0‐OOH(‐O2−) (i.e., the DMPO spin adduct of ‐O2−) was strongest for 2‐amino‐3,4‐dimethy]imidazo[4,5‐f]quinoline (MeIQ). The O2− generation with HCAs decreased in the following order: 2‐amino‐3,8‐dimethyI‐imidazo[4,5‐f] quinoxaline (MeIQx) = 2‐amino‐3‐methylimidazo[4,5‐f]quinoline (IQ) > 2‐amino‐3,4,8‐trimethylimidazo[4,5‐f]quinoxaline (diMeIQx) ≥ other HCAs; O2− generation was lowest with 2‐amino‐3‐methyl‐9H‐pyrido[2,3‐b]indole CH3COOH (MeAαC). By using Lineweaver‐Burk plots, Km values of cytochrome P‐450 reductase for mitomycin C., IQ, and MeIQ were determined to be 1,60 × 10−6 M, 1.97 × 10−5M, and 2.83 × 10−6 M, respectively. The present findings have important implications for carcinogenesis because of the known effect of oxygen radicals on cell proliferation.
Quercetin, an Inhibitor of Heat Shock Protein Synthesis, Inhibits the Acquisition of Thermotolerance in a Human Colon Carcinoma Cell Line
Authors: Koishi Mototsugu , Hosokawa Nobuko , Sato Mamoru , Nakai Akira , Hirayoshi Kazunori , Hiraoka Masahiro , Abe Mitsuyuki , Nagata Kazuhiro
Published: August 2005

Here, we describe the effects of quercetin on the induction of thermotolerance as examined by colony forming assay in a cell line derived from human colon carcinoma (COLO320 DM). Cells became resistant to heat treatment at 45°C when they were preheated at 42°C for 1.5 h or at 45°C for 10 min. This induction of thermotolerance was almost completely inhibited by continuous treatment with 100 μM quercetin during the first and second heating sessions, and the interval between. This effect of quercetin was demonstrated to be dose‐dependent over a concentration range of 50 200 μM, Quercetin did not increase the thermosensitivity of non‐tolerant cells. The presence of quercetin during the first conditioning heating was more effective in inhibiting thermotolerance than its presence during the second heating. Quercetin was also found to inhibit the acquisition of thermotolerance induced by sodium arsenite. Cycloheximide, a nonspecific inhibitor of protein synthesis, did not affect the acquisition of thermotolerance by the same cell line, Quercetin specifically inhibits the synthesis of all heat shock proteins so far reported previously, and this leads to inhibition of the induction of thermotolerance. Such inhibition of thermotolerance by quercetin may improve the efficacy of clinical fractionated hyperthermia.
Immunohistochemical Localization of Hepatocyte Growth Factor Protein in Pancreas Islet A‐Cells of Man and Rats
Authors: Tsuda Hiroyuki , Iwase Teruhiko , Matsumoto Kazuyuki , Ito Mitsuya , Hirono Iwao , Nishida Yoshihisa , Yamamoto Masami , Tatematsu Masae , Matsumoto Kunio , Nakamura Toshikazu
Published: August 2005

Hepatocyte growth factor (HGF), a potent mitogen for adult rat hepatocytes in primary culture, has previously been shown to be primarily expressed in the nonparenchymal cells of the liver. Using polyclonal antisera against human and rat HGFs we studied the tissue distribution of HGF immunohistochemically and found the most intense staining in the pancreas islet cells in both man (autopsy cases) and the rat. Differential localization of 4 pancreas islet hormones, glucagon, insulin, somatostatin and pancreatic polypeptide, revealed HGF to be preferentially expressed within the glucagon‐positive cells. The results indicate that HGF is primarily produced or stored in A‐cells and may act as a growth factor in a paracrine and an endocrine fashion, like various other hormones.
The Effects of Sodium Chloride, Miso or Ethanol on Development of Intestinal Metaplasia after X‐Irradiation of the Rat Glandular Stomach
Authors: Watanabe Hiromitsu , Okamoto Taro , Takahashi Tadateru , Ogundigie Peter Osa , Ito Akihiro
Published: August 2005

The influence of sodium chloride (NaCl), miso (Japanese soybean paste) and ethanol on development of intestinal metaplasia was examined. Five‐week‐old male CD(SD): Crj rats were treated with two 10 Gy doses of X‐rays to the gastric region at a 3‐day interval (total 20 Gy). After irradiation, the rats received supplementation with NaCl (1% or 10% in diet), miso (10% in diet) or ethanol (10% in drinking water) for 12 months. The number of alkaline phosphatase‐positive foci of intestinal metaplasia in rats given 1% NaCl diet (Group 3) after X‐rays was significantly elevated as compared to that in rats given X‐rays alone (Group 1) (P < 0.01) or X‐rays with 10% NaCl (Group 2) (P < 0.01). In the pyloric gland mucosae, the total numbers of metaplastic foci in rats of Group 3 were much higher than in Group 2, or after miso diet (Group 4) or ethanol supplementation (Group 5) (P < 0.01), but no difference was found between Group 2, 4 or 5 and Group 1. Atypical hyperplasia only appeared at incidences of less than 6% in Groups 1–3 and no promoting effect on gastric tumorigenesis was evident in Group 2. The present results thus showed that the occurrence of intestinal metaplasia induced by X‐irradiation can be significantly increased by administration of 1% NaCl and decreased by 10% NaCl and ethanol, but this is not associated with any influence on gastric neoplasia.
Modifying Effects of Fungal and Herb Metabolites on Azoxymethane‐induced Intestinal Carcinogenesis in Rats
Authors: Yoshimi Naoki , Wang Aijin , Morishita Yukio , Tanaka Takuji , Sugie Shigeyuki , Kawai Kiyoshi , Yamahara Joji , Mori Hideki
Published: August 2005

Modifying effects of a fungal product, flavoglaucin, and four plant‐derived chemicals, shikonin, gingerol, oleanolic acid and paeoniflorin, on intestinal carcinogenesis were examined in a rat model using azoxymethane (AOM). A total of 280 male F344 rats, 6 weeks old, were divided into 12 groups. Group 1 (30 rats) was given two subcutaneous injections of 15 mg/kg of AOM at the start of the experiment. Groups 2 (30 rats), 3 (20 rats), 4 (20 rats), 5 (30 rats) and 6 (30 rats) received a test chemical (flavoglaucin, shikonin, gingerol, oleanolic acid or paeoniflorin, respectively) in the diet at a concentration of 0.02% for 3 weeks, during which time AOM was applied, and then kept on basal diet until the end of experiment (one year). Groups 7–11 (each 20 rats) were given a test chemical corresponding to Groups 2–6, respectively. Group 12 (20 rats) served as a control. The incidence and average number of intestinal tumors in Group 2 (47%, 0.57 ± 0.68) were significantly less than in Group 1 (74%, 1.07 ± 0.87) (P < 0.05, respectively). Multiplicity of intestinal neoplasms of Group 3 (0.55 ± 0.60) or 4 (0.47 ± 0.51) was also significantly smaller than that of Group 1 (P < 0.05 and P < 0.01, respectively). These results suggest that flavoglaucin, shikonin and gingerol might be promising chemopreventive agents for intestinal neoplasia.
p53 Gene Mutations Associated with Anaplastic Transformation of Human Thyroid Carcinomas
Authors: Nakamura Tsutomu , Yana Ikuo , Kobayashi Tetsuro , Shin Eisei , Karakawa Katsu , Fujita Shoichi , Miya Akihiro , Mori Takesada , Nishisho Isamu , Takai Shin‐ichiro
Published: August 2005

Anaplastic carcinoma of the thyroid gland, which is one of the most aggressive, malignant tumors in humans, is considered to originate from preexisting differentiated thyroid cancer. To define the genetic alterations associated with such progression, we examined nine cases of anaplastic thyroid carcinoma for mutation in exons 4–9 of the p53 tumor suppressor gene. Preliminary screening for mutation by RNase protection analysis demonstrated that two out of nine anaplastic carcinomas contained sequence alterations in the p53 gene. Subsequent DNA sequencing identified the mutated nucleotides in these two cases; one was a nonsense mutation at codon 165, and the other was a single‐base deletion at codon 176 resulting in the creation of a stop codon downstream due to frameshift. The fact that no mutations were detected in coexisting foci of papillary carcinomas from the same patients shows that these mutations of the p53 gene occurred after development of papillary carcinomas. These results suggest that p53 gene mutation triggers the progression from differentiated into anaplastic carcinoma in the human thyroid gland.
Inhibition of Cell Attachment, Invasion and Metastasis of Human Carcinoma Cells by Anti‐integrin β1 Subunit Antibody
Authors: Fujita Shin , Suzuki Hiroko , Kinoshita Masao , Hirohashi Setsuo
Published: August 2005

We investigated the expression of β1 integrins in human carcinoma cell lines, and the anti‐metastatic and anti‐invasive effects of a newly established anti‐human β1 subunit monoclonal antibody designated NCC‐INT‐7. All the examined carcinoma cell lines expressed β1 integrins upon immunoblot analysis. NCC‐INT‐7 completely inhibited the adhesion of carcinoma cells to laminin, fibronectin, collagens and acetone‐fixed tissues including lung, liver and brain. In an in vitro invasion model, NCC‐INT‐7 inhibited the invasion of human bladder carcinoma cell line T24 and human gastric carcinoma cell lines TMK‐1, MKN‐45 and MKN‐74 through an artificially reconstructed basement membrane. In an in vivo nude mouse peritoneal dissemination model using MKN‐45 and TMK‐1, NCC‐INT‐7 significantly reduced the number of tumor nodules in the mesentery. In an in vivo nude mouse liver metastasis model using a serially transplantable human colonic carcinoma, COL‐2–JCK, NCC‐INT‐7 significantly reduced the number of tumor nodules in liver. These results indicate that β1 integrins play an important role in the tissue attachment, migration, invasion and metastasis of human carcinoma cells, and that this new monoclonal antibody is useful for studies aimed at prevention of metastasis.
Glomeruloid Structures as Vascular Reaction in Human Gastrointestinal Carcinoma
Authors: Ohtani Haruo
Published: August 2005

Malignant tumors induce angiogenesis and modulation of microvasculature. Based on histologic and immunohistochemical analysis of human surgical material, we describe here the occurrence of glomeruloid structures in gastrointestinal carcinomas, and compare them with the microvasculature in inflammatory granulation tissue. The glomeruloid structures were composed of clusters of mutually fused capillaries with prominent swelling of endothelial cells and pericytes. They were thought to be specific for glioblastoma of the brain. The glomeruloid structures were observed juxtaposed to carcinoma nests in one‐third of gastric carcinoma of intestinal type and colorectal carcinoma in the area of invasive growth beyond the muscularis mucosae. They were not observed in gastric carcinoma of diffuse type, intramucosal carcinoma, or inflammatory granulation tissue. The glomeruloid structures can be regarded as an extreme example of endothelial hyperplastic changes observed in cancer stroma. Our results suggested that glomeruloid structures can occur in carcinomas as vascular reaction, a mechanism different from that in inflammatory granulation tissues.
Effect of Recombinant Human D‐Factor on the Growth of Leukemic Blast Progenitors from Acute Myeloblastic Leukemia Patients
Authors: Chen Guan‐Jin , Tomida Mikio , Hozumi Motoo , Nara Nobuo
Published: August 2005

We studied the effects of D‐factor on the growth of leukemic blast progenitors from 15 patients with acute myeloblastic leukemia and two leukemia cell lines in methylcellulose and suspension cultures. When stimulated by granulocyte colony‐stimulating factor (G‐CSF), granulocyte‐macrophage colony‐stimulating factor or interleukin‐3, leukemic blast progenitors undergo terminal division with limited differentiation in methylcellulose culture, forming blast colonies. Leukemic blast progenitors can renew themselves. The self‐renewal can be detected as secondary colony formation after replating primary blast colonies in fresh methylcellulose media and by the growth of clonogenic cells in suspension culture. D‐Factor suppressed primary and secondary colony formation in methylcellulose culture. Furthermore, D‐factor suppressed clonogenic cell recovery in suspension culture. The suppression by D‐factor of the growth of leukemic blast progenitors was not significantly dependent upon the colony‐stimulating factors used as growth‐stimulating factors. High concentration of G‐CSF did not overcome the suppressive effect of D‐factor. The results indicate that D‐factor is effective in suppressing not only terminal division but also self‐renewal of leukemic blast progenitors.
Analysis by Step Sectioning of Early Invasive Bladder Cancer with Special Reference to G3·pT1 Disease
Authors: Kakizoe Tadao , Tobisu Kenichi , Mizutani Takashi , Tsutsumi Masakazu , Tanaka Yoshinori , Sakamoto Michi‐ie
Published: August 2005

Eighty cases of cystectomized and step‐sectioned pT1 transitional cell carcinomas of various grades were analyzed. During the same period, 30 consecutive cases of pT2 carcinomas were also cystectomized and examined for comparison. This is a cross‐sectional study of a variety of cases of bladder cancer, designed to delineate the characteristics of G3·pT1 disease. Tumors in this series had a full set of various pathological findings; grades 1 to 3, stages Ta‐T1‐T2, papillary‐papillonodular‐nodular tumor configuration, α‐β‐γ type of invasion, presence of lymphatic and venous involvement, and presence of associated carcinoma in situ/ dysplasia. There is a distinct tendency of stepwise disease progression, such as grade 1→3, stage T1→T2, papillary→nodular configuration, and α→γ invasion, these factors being mutually related. We noted a similarity between tumor groups containing G3 component, such as G2‐3·pT1, G3 >2 ·pT1, G3·pT1, G1‐3·pT2, and G3·pT2. These tumors are different from groups such as G1‐2·pT1, G2·pT1 and G2 >3·pTl in terms of tumor configuration and type of invasion. As regards pT1 having G3 components, early cystectomy seems to be the surest treatment. Whenever one adopts a conservative policy in treatment of these tumors, extreme care should be taken to monitor tumor progression.
Solid‐phase Anti‐CD3 Antibody Activation and Cryopreservation of Human Tumor‐infiltrating Lymphocytes Derived from Epithelial Ovarian Cancer
Authors: Ikarashi Hirokazu , Aoki Yoichi , Fujita Kazuyuki , Kodama Shoji , Tanaka Kenichi
Published: August 2005

The effect of solid‐phase anti‐CD3 antibody activation and cryopreservation was evaluated on thirteen samples of tumor‐infiltrating lymphocytes (TILs) derived from epithelial ovarian cancer. Seven preparations of TILs were cultured with or without solid‐phase anti‐CD3 antibody in addition to 100 units/ml of recombinant interleukin‐2 (rIL‐2). The proliferation rate of all of the seven TIL preparations stimulated by anti‐CD3 antibody on the fourth or fifth day of culture was 3.4 to 9.8 times greater than that of lymphocytes cultured with rIL‐2 alone. Furthermore, in an experiment with five TIL samples activated with anti‐CD3 antibody, three of them showed augmented cytotoxic activity against autologous fresh tumor cells. The population of CD3+/CD8+ TILs was increased after 4–5 weeks of cultivation and CD8+ lymphocytes amounted to over 70% in all of seven preparations tested, whereas two of seven preparations not activated by anti‐CD3 antibody were CD3+/CD4+‐dominant. In addition, nine preparations of TILs cultured with rIL‐2 were cryopreserved for several weeks; after recovery from cryopreservation, no major change was observed in cell surface markers, in growth rate or in cytotoxic activity. These results suggest that cryopreserved and/or anti‐CD3 antibody‐activated lymphocytes could conveniently be employed in a clinical trial of adoptive immunotherapy employing TIL.
Establishment of an Enzyme Immunoassay Using Monoclonal Antibody (HG1‐219) and Its Application for the Diagnosis of Hepatocellular Carcinoma
Authors: Hada Toshikazu , Kinoshita Yumiko , Ichimori Yuzo , Imanishi Hiroyasu , Umeyama Keishi , Amuro Yoshiki , Toyosaka Akihiro , Okamoto Eizo , Higashino Kazuya
Published: August 2005

A monoclonal antibody (MoAb HG1‐219) against a human gastric cancer cell line (HuG‐1) and its shedding antigen (HG1‐219 Ag) was generated and a solid‐phase sandwich enzyme immunoassay (EIA‐219) was developed. The mean serum HG1‐219 Ag concentration in normal individuals was 30.5 ± 14.5 U/ml measured by EIA‐219. When the mean + 3 SD of the antigen concentration in normal individuals was used as a cut‐off level, 4.3% (2/47) of patients with chronic hepatitis, 9.1% (4/44) of cirrhotic patients and 37.5% (18/48) of patients with hepatocellular carcinoma (HCC) had HG1‐219 Ag above the cut‐off value. The positive rates of a‐fetoprotein (AFP) (> 400 ng/ml) and protein induced by vitamin K absence or antagonist‐II (PIVKA‐II) for HCC were 26.7% (12/45) and 33.3% (12/36), respectively. There was no significant correlation between HG1–219 Ag and AFP or PIVKA‐II in patients with HCC. The combination assay of EIA‐219, AFP and PIVKA‐II for HCC gave the positive rate of 75% (27/36). The effect of periodic acid on the HG1‐219 Ag and the inhibition of EIA‐219 by CA 19–9 suggest that the epitope of HG1–219 Ag is a suger chain similar to CA 19–9.
Immunohistochemical Type Distinction of a‐Fetoprotein in Various a‐Fetoprotein‐secreting Tumors
Authors: Okamoto Tsukasa , Hirabayashi Koji , Ishiguro Tatsuya
Published: August 2005

In order to clarify the histogenesis of a‐fetoprotein(AFP)‐secreting tumor tissues, formalin‐fixed, paraffin‐embedded serial sections of 148 tumors in various organs were examined by the peroxidase‐antiperoxidase method for AFP, and paradoxical concanavalin A staining. Yolk sac‐type AFP was found in yolk sac tumors, embryonal carcinomas, solid teratomas, (yolk sac) endodermal cell tumors, adenocarcinomas (stomach, ovary or lung) and metastatic liver cancers. Hepatic‐type AFP was demonstrated in hepatocellular carcinomas, hepatoblastomas, solid teratomas and a stomach cancer. Yolk sac‐type AFP was observed in the neighboring liver cells of metastatic liver cancers without relation to the type of AFP in primary cancers. The results from serum analyses of preoperative tumor‐bearing patients (68 cases) were coincident with those from immunohistochemical stainings.
Novel Interleukin‐2 Dependent T‐Cell Line Derived from Adult T‐Cell Leukemia Not Associated with Human T‐Cell Leukemia Virus Type I
Authors: Kagami Yoshitoyo , Tobinai Kensei , Kinoshita Tomohiro , Nagai Hirokazu , Hotta Tomomitsu , Seto Masao , Ueda Ryuzo , Saito Hidehiko
Published: August 2005

A novel interleukin‐2 (IL‐2)‐dependent T‐cell line, WHN2, was established from a patient with adult T‐cell leukemia (ATL) not associated with human T‐cell leukemia virus type I (HTLV‐I). Neither the original leukemic cells nor the WHN2 cells showed proviral integration in their cellular DNAs by Southern blot analysis. The surface phenotype showed that both the original leukemic cells and the WHN2 cells had a common phenotype of ATL, i.e., positive for CD2, CD4, human leukocyte antigen DR (HLA‐DR) and CD25, but negative for CD8, a characteristic of helper/inducer T‐cells. Most of the chromosomal abnormalities of the original leukemic cells were maintained in the WHN2 cell line. Furthermore, Southern blot analysis of the T‐cell receptor β‐chain gene rearrangement revealed that the original leukemic cells and WHN2 cell line had identical patterns, suggesting that the WHN2 cell line was truly derived from the original leukemic cells. Dose‐dependent growth on IL‐2 was demonstrated, and at the maximal stimulation, the number of cells doubled within three days. This IL‐2‐dependent growth was inhibited by the simultaneous existence of anti‐IL‐2 receptor a and β chain antibodies. These results indicate that the character of the WHN2 cell line is similar to that of the cell lines derived from ATL associated with HTLV‐I. Thus, the HTLV‐I‐negative ATL cell line, WHN2, should be useful in the comparative study of the pathogenesis of ATL associated with or without HTLV‐I.
Clonal Analysis of Multiple Point Mutations in the N‐ras Gene in Patients with Acute Myeloid Leukemia
Authors: Kubo Kazuaki , Naoe Tomoki , Kiyoi Hitoshi , Fukutani Hisashi , Kato Yoshiro , Oguri Takashi , Yamamori Shunji , Akatsuka Yoshiki , Kodera Yoshihisa , Ohno Ryuzo
Published: August 2005

We have screened mutations of the N‐ras gene at codons 12, 13, and 61 in leukemia cells obtained from 100 patients with acute myeloid leukemia (AMD, and found mutated N‐ras alleles in 9 patients. We further analyzed the polyclonality of multiple N‐ras gene mutations in 4 AML patients. One patient, who had the monoclonal karyotype, t(11;17), had two types of double missense mutations at codons 13 and 61 in the same allele. Each of the remaining three patients, one of whom had t(15;17) with a monoclonal rearrangement of the retinoic acid receptor alpha and PML genes, carried two missense mutations in a relatively small population of leukemia cells. We have demonstrated that multiple clonality of the N‐ras gene is occasionally observed in leukemia with a monoclonal karyotype. These findings indicate that the N‐ras mutations may not always be characterized simply by an accumulative process and that the activated N‐ras gene alone is not sufficient to cause leukemia.
p53 Mutations and c‐erbB‐2 Amplification in Intraductal and Invasive Breast Carcinomas of High Histologic Grade
Authors: Tsuda Hitoshi , Iwaya Keiichi , Fukutomi Takashi , Hirohashi Setsuo
Published: August 2005

In order to clarify the clinical significance of mutations of the p53 gene and amplification of the c‐erbB‐2 gene in breast carcinoma, these gene alterations were examined in 101 invasive, seven predominantly intraductal and 10 intraductal breast carcinomas by single‐strand conformation polymorphism‐direct sequencing or Southern blot‐hybridization analysis. p53 mutations were detected in 32 (32%) of the invasive cases and two (12%) of the 17 intraductal/predominantly intraductal cases, whereas c‐erbB‐2 amplification was detected in 14 (14%) of the invasive and six (35%) of the intraductal/predominantly intraductal cases. Irrespective of differences in the positions and types of the mutations, cases carrying p53 mutations were almost always Grade 3 histologically and with a low hormone‐receptor value. Since p53 mutations as well as c‐erbB‐2 amplification were detected almost selectively in Grade 3 cases but were not associated with lymph nodal status in invasive breast cancer, these two gene alterations could be indicators of prognosis of disease independent of lymph nodal status. Even in intraductal/predominantly intraductal carcinoma, these gene alterations were almost always detected in tumors of higher histologic grade. Thus, it is suggested that these gene alterations occur in breast cancers showing a high proliferation rate irrespective of the presence of invasion, and that other molecular alterations are involved in the process of breast cancer invasion.
Deletion Mapping of Chromosome 1p and 22q in Pheochromocytoma
Authors: Shin Eisei , Fujita Shoichi , Takami Koji , Kurahashi Hiroki , Kurita Yoshihiro , Kobayashi Tetsuro , Mori Takesada , Nishisho Isamu , Takai Shin‐ichiro
Published: August 2005

To identify the localization of tumor suppressor genes, 22 pheochromocytomas (9 hereditary and 13 sporadic) were examined for loss of heterozygosity (LOH) on the short arm of chromosome 1 and on the long arm of chromosome 22 by using 11 polymorphic DNA markers on each chromosome arm. LOH on 1p was observed in 12 of 22 informative cases (55%) and on 22q in 8 of 20 informative cases (40%). There was no significant difference in the frequency of LOH on 1p or 22q between hereditary and sporadic cases. We could localize the commonly deleted regions as distal to D1S73 and proximal to D1S63 on 1p and distal to D22S24 and proximal to D22S1 on 22q. In addition, the relationship between LOH on 1p and 22q was studied in 20 pheochromocytomas which were informative for probes on both chromosome arms. Of eight tumors that showed LOH on 22q, allelic loss on 1p was also detected in seven. Thus, LOH on 22q was correlated significantly with LOH on 1p (P= 0.0249; Fisher's exact test). These results suggest that inactivation of multiple tumor suppressor genes may be required for development and progression of hereditary and non‐hereditary pheochromocytoma.
Enhancing Effect of Interleukin‐2 on Production of Parathyroid Hormone‐related Protein by Adult T‐Cell Leukemia Cells
Authors: Mori Naoki , Ohsumi Kazuoki , Murakami Shuichi , Wake Atsushi , Shirakawa Fumihiko , Morimoto Isao , Oda Susumu , Eto Sumiya
Published: August 2005

Leukemic cells from patients with adult T‐cell leukemia (ATL) can produce a calcium‐regulating protein, parathyroid hormone‐related protein (PTHrP). Moreover, it has been reported that ATL cells produce some cytokines besides PTHrP and that these cells respond to the T‐cell growth factors, interleukin‐2 (IL‐2) and interIenkin‐4 (IL‐4). To elucidate whether PTHrP produced by ATL cells is regulated by IL‐2 or IL‐4, we investigated the in vitro effects of IL‐2 and IL‐4 on the release of PTHrP. IL‐2 increased the release of PTHrP into the conditioned medium from leukemic cells in some, but not all, ATL patients; however, IL‐4 did not affect the PTHrP release. PTHrP messenger RNA (mRNA) levels were increased in ATL cells cultured in the presence of IL‐2. These data suggest that IL‐2 plays a role in the regulation of hypercalcemia by enhancing the production of PTHrP in ATL patients.
Interleukin‐2 Prevents Programmed Cell Death in Adult T‐Cell Leukemia Cells
Authors: Tsuda Hiroyuki , Huang Ren‐Wei , Takatsuki Kiyoshi
Published: August 2005

Adult T‐cell leukemia (ATL) is a prototype of the lymphoma/leukemia syndromes involving immunologically mature T‐lymphocytes. The first retrovirus described in humans, HTLV‐I, is causally related to the disease. In this study, we examined whether ATL cells die in vitro through programmed cell death (PCD), which has been shown to occur in cells affected by several other acute and chronic Icukcmias. When ATL cells from peripheral blood were cultured in serum‐free complete medium, a substantial proportion of them spontaneously died by PCD. After 48 h of culture, approximately 30% of the total DNA was fragmented. Electrophoresis indicated that the DNA of the ATL cells had been cleaved into regular oligonucleosome fragments each comprising approximately 180–200 base pairs. This process was significantly promoted by methylprednisolone and the protein kinase A (PKA) activator Sp‐cAMPS in at least some cases. Since all ATL cells possess interleukin‐2 receptors on the cell membrane, the effect of IL‐2 on spontaneous PCD was assessed. PCD after 48 h of culture was inhibited by 30–50% by 100 U/ml interleukin‐2 (IL‐2). This effect of IL‐2 to prevent spontaneous PCD was dose‐ and time‐dependent. These findings suggest that the viability of ATL cells in vivo is regulated positively and negatively by intrinsic IL‐2, glucocorticoid and regulators of PKA activity. Furthermore, the process of cell death may be involved in the development of the disease.
Effect of Accessory Cells on Stimulation of Murine T‐Cell Leukemia with Antibodies to the CD3/T Cell Antigen Receptor Complex
Authors: Suto Ryuichiro , Udono Heiichiro , Yamamoto Akihito , Shiku Hiroshi , Nakayama Eiichi
Published: August 2005

Stimulation of EL4 and RL1 leukemia cells in vitro with immobilized anti‐CD3ε monoclonal antibody (mAb) (145–2C11) or anti‐TCRβ mAb (H57–597) in the absence of accessory cells induced interleukin‐2 (IL‐2) production, and caused growth inhibition. The growth inhibition was, however, transient and the tumors started to grow again within 5 days in immobilizing plates treated with antibodies at concentrations of 2.5–100 μg/ml. Addition of mitomycin C‐treated accessory cells to the culture inhibited IL‐2 production and resulted in augmented and persistent growth inhibition. No recovery of tumor growth was observed. Furthermore, DNA from EL4 and RL1 leukemia cells stimulated with anti‐CD3/TCR mAbs was fragmented even in the absence of accessory cells, but fragmentation was much greater in the presence of accessory cells. Marginal and high expression of the bcl‐2 gene were observed in EL4 and RL1, respectively, indicating that apoptosis of these leukemias mediated by signalling through the CD3/TCR complex has no direct relationship with expression of the bcl‐2 gene.
Lysosome Labilizers Potentiate the Antitumor Effects of Tumor Necrosis Factor‐α
Authors: Masegi Tsukio , Kato Arata , Kitai Kazuo , Fukuoka Masami , Soma Kazuhiko , Ichikawa Yataro , Nakamura Satoshi , Watanabe Naoki , Niitsu Yoshiro
Published: August 2005

Enhancement of in vitro cytotoxic activity of tumor necrosis factor‐α (TNF‐α) was observfed in combination with lysosome labilizers, particularly with urokinase‐type plasminogen activator (u‐PA), tissue‐type plasminogen activator (t‐PA) and lipoprotein lipase (LPL). The concentration of TNF‐α resulting in 50% cytotoxicity to L929 cells was only 20–30% of the value for TNF‐α alone, when used in combination with a nontoxic dose of u‐PA, t‐PA or LPL. Furthermore, combined intravenous (i.v.) administration of TNF‐α (3.5 × 105 U/mouse) and u‐PA (300 IU/mouse) markedly increased the in vivo antitumor activity of TNF‐α to Meth A tumors transplanted into BALB/c mice; the tumor weight in co‐administered mice was about 40% of that in mice given TNF‐α alone on day 6. The combination therapy of TNF‐α (7.0 × 104 U/mouse, i.v.) and u‐PA (300 IU/mouse, i.v.) was also effective for L929 tumors in Crj:CD‐1(1CR)‐nu nude mice compared with the conventional therapy with TNF‐α alone. These results suggest that the combination of TNF‐α and lysosome labilizers is a promising antitumor therapeutic regimen with clinical potential.
Inhibition of Cisplatin‐mediated DNA Damage in vitro by Ribonucleotides
Authors: Seki Shuji , Hongo Atsushi , Zhang Bo , Akiyama Kosuke , Sarker Altaf H. , Kudo Takafumi
Published: August 2005

Effects of ribonucleotides on cis‐diamminedichloroplatinum(II) (cisplatin)‐mediated DNA damage were studied by incubating pUC18 DNA with cisplatin in the presence of nucleotide, and by monitoring conformational change and sensitivity change to restriction enzyme HpaII of the DNA due to the platinum‐DNA adduct formation. The cisplatin‐mediated DNA damage was inhibited in a dose‐dependent fashion by ATP or GTP, substantially at their physiological intracellular concentrations, and almost completely by 5 mM ATP or 2 mM GTP. The inhibitory effect of nucleotide on the platination of DNA was in the order of GTP > ATP≫CTP > UTP, and of nucleoside triphosphate >nucleoside diphosphate > nucleoside monophosphate. Nucleoside did not show any significant effect on platination of DNA. To elucidate the mechanism of the nucleotide effects on platination of DNA, interaction between ATP or GTP and cisplatin was analyzed by high‐performance liquid chromatography. The results suggested that ATP inhibits cisplatin‐mediated DNA damage both by forming a platinum‐ATP adduct and by non‐covalent ionic interaction with cisplatin, while GTP acts largely by forming platinum‐GTP adducts.
Effect of Experimental Hypochlorhydria on the Histological Differentiation of Adenocarcinomas in the Canine Stomach
Authors: Sano Takeshi , Kobori Oichiro , Kuroki Sho , Nagawa Hirokazu , Muto Tetsuichiro
Published: August 2005

Hypochlorhydria is considered to be a risk factor for gastric cancer, both clinically and experimentally. In humans, the hypo‐acidic condition appears to be closely associated with the development of differentiated‐tyne gastric carcinomas. We investigated relationships between antral pH and histological types of gastric cancer using an animal model. A total of 7.65 g of N‐ethyl‐N′‐nitro‐N‐nitrosoguanidine (ENNG) was orally administered to male beagle dogs. Subsequently, the dogs were divided into four groups and underwent four surgical interventions designed to create various conditions in the antrum. Antral pM was recorded at the time of killing after an observation period ranging from 16 to 33 months. The number of gastrin‐secreting cells (G‐cells) was counted after immunohistochemical staining. Carcinomas were found in the antrum of 12 of the 21 dogs that had survived. Although limited doses of ENNG, such as those used in this study, have been reported to produce only undifferentiated adenocarcinomas, differentiated adenocarcinomas were found in seven of these 12 dogs. Antral pH and the number of G‐cells were significantly higher in these seven dogs than in the other five dogs, which had developed only undifferentiated adenocarcinomas. Bile inflow showed no significant effect on the development of differentiated adenocarcinomas. Neither atrophic gastritis nor intestinal metaplasia was seen in any of the dogs. These results suggest that the hypo‐acidic condition itself may play a role in the development of differentiated adenocarcinomas.
Benign Breast Disease as a Breast Cancer Risk in Japanese Women
Authors: Nomura Yasuo , Tashiro Hideya , Katsuda Yasaburo
Published: August 2005

A hospital‐based retrospective cohort study of benign breast disease (BBD) as a risk factor of future breast cancer (BC) development was conducted. Four hundred and twenty‐eight patients with biopsied BBD were followed‐up for a median period of 8 years, together with age‐matched women with normal breasts (normal control) and BC patients (cancer control), at the ratio of 1:2:2. Twenty‐one breast cancers developed, 7 in the cases, 4 in the normal controls, and 10 in the BC controls, showing the relative risk (RR) with 95% confidence intervals (CD to be 3.5 (1.03–11.9) in the cases with respect to the normal controls. The RR of the cases is not lower than that of contralateral breast cancer incidence. There were no significant differences in the risks of cancers in other organs among the groups. Pathological examination revealed that only atypical hyperplasia increased the RR of BC, as compared with the normal control breast group, or with non‐proliferative disease. These results suggest that in a low‐risk country, Japan, BBD is a definite risk factor for BC development as in high‐risk countries.
Enhancing Effects of Calcium‐deficient Diet on Gastric Carcinogenesis by N‐Methyl‐N′‐nitro‐N‐nitrosoguanidine in Wistar Rats
Authors: Tatsuta Masaharu , Iishi Hiroyasu , Baba Miyako , Uehara Hiroyuki , Nakaizumi Akihiko , Taniguchi Haruo
Published: August 2005

The effects of ad libitum feeding of calcium‐deficient diet on the incidence, number and histological types of gastric cancers induced by N‐methyl‐N′‐nitro‐N‐nitrosoguanidine (MNNG) were investigated in male Wistar rats. Rats were fed standard pellet diet containing 0.5% (normal‐calcium diet) or 0.01% calcium (calcium‐deficient diet) after oral treatment with MNNG for 25 weeks. Oral administration of the calcium‐deficient diet resulted in a significant increase in the incidence, hut not the number, of gastric cancers in experimental Week 52. However, it did not affect the histological types of cancer. The calcium‐deficient diet also caused a significant increase in tissue norepinephrine concentration of the antral portion of the gastric wall and in the labeling index of the antral epithelial cells. These findings indicate that the calcium‐deficient diet enhanced gastric carcinogenesis and suggest that its effect may be related to increase in norepinephrine in the gastric wall and consequent stimulation of proliferation of antral epithelial cells.
Comparison of K‐ras Oncogene Activation in Pancreatic Duct Carcinomas and Cholangiocarcinomas Induced in Hamsters by N‐Nitrosobis(2‐hydroxypropyl)amine
Authors: Tsutsumi Masahiro , Murakami Yoshinori , Kondoh Satoshi , Tsujiuchi Toshifumi , Hohnoki Kanya , Horiguchi Kohsuke , Noguchi Osamu , Kobayashi Eisaku , Okita Shunji , Sekiya Takao , Konishi Yoichi
Published: August 2005

The presence of K‐ras point mutations in pancreatic duct carcinomas and Cholangiocarcinomas induced by N‐nitrosobis(2‐hydroxypropyl)amine (BHP) in Syrian hamsters was investigated by single‐strand conformation polymorphism analysis of polymerase chain reaction products from frozen fresh materials in order to clarify the K‐ras mutation rates in those two carcinomas induced simultaneously by one carcinogen, BHP. In the examined pancreatic duct carcinomas, 10 out of 16 were positive for a mutation in codon 12 while 3 out of 12 Cholangiocarcinomas demonstrated mutation of K‐ras gene. G‐to‐A transition was detected in the second position of codon 12 in both pancreatic carcinomas and Cholangiocarcinomas. These results suggest that the role of genetic alteration in carcinogenesis may differ with the target organ, even when initiation is with the same carcinogen.
Follow‐up Study of Patients Treated with Monoclonal Antibody‐Drug Conjugate: Report of 77 Cases with Colorectal Cancer
Authors: Takahashi Toshio , Yamaguchi Toshiharu , Kitamura Kazuya , Noguchi Akinori , Honda Mitsuyo , Otsuji Eigo
Published: August 2005

A total of 77 patients with advanced colorectal cancer, including postoperative patients with liver, lung and peritoneal metastases, were treated with single or multiple injections of monoclonal antibody A7‐neocarzinostatin (A7‐NCS). A follow‐up study of the patients treated with A7‐NCS was done and the clinical outcome was compared with that of patients given other chemotherapies. In the postoperative patients with liver metastasis, the A7‐NCS treatment prolonged survival time when compared with systemic administration of anticancer drugs, while it showed a similar survival time to chemoembolization using multiple anticancer agents suspended in a lipid contrast medium. Among the patients who underwent surgical resection of primary cancer, with or without liver metastasis, there was no difference in overall 5‐year survival rate between the group treated with A7‐NCS and the group treated with the other chemotherapies. However, the survival time of the patients treated with A7‐NCS was longer than that of the patients treated with the other chemotherapies. In addition, the patients given a higher dose of A7‐NCS had a longer survival time than the patients given a lower dose of A7‐NCS. Human anti‐mouse antibody was detected in all the A7‐NCS‐treated patients examined. There were no serious side effects in any of the patients given A7‐NCS. Thus, this study indicates that the A7‐NCS treatment is safe and useful for colorectal cancer patients, though some problems remain, such as optimization of injection dose, route, interval, etc., and overcoming human anti‐mouse antibody development.
Aberrant α1→2Fucosyltransferases Found in Human Colorectal Carcinoma Involved in the Accumulation of Leb and Y Antigens in Colorectal Tumors
Authors: Yazawa Shin , Nakamura Jun‐ichi , Asao Takayuki , Nagamachi Yukio , Sagi Morihisa , Malta Khushi L. , Achikawa Tetsuya T , Akamatsu Masaru
Published: August 2005

Evidence indicates that the presence of aberrant α1→2fucosylation pathways is responsible for the accumulation of large quantities of Leb and Y antigens in human colorectal carcinoma. Significantly higher activities of α1→2 as well as α1→3 and α1→4fucosyltransferases were found in most of the tissues from carcinoma than in the adjacent normal tissues and in healthy subjects. α1→2Fucosyl‐transferases associated with the synthesis of Leb (Fucal→2Galβ1→3[Fucc1→4]GlcNAcβ) and Y (Fucα1→2Ga1β→4[Fucα1→3]GlcNAcβ) structures from Le→ (GaIβ1→3[Fucal→4]GlcNAcβ) and X (Galβ1→4[Fucα 1→3]GlcNAcβ) ones, respectively, were demonstrated in colorectal carcinomas and in colorectal carcinoma cell lines (COLO201, LS174T and SW1116). The activation of α1→2fucosyltransferase with such new substrate specificities in colorectal carcinoma might result in the preferential synthesis of Leb and Y structures from Le→ and X rather than from H type 1 and H type 2 structures.
Potentiation of Metastatic Capacity by Transforming Growth Factor‐β1 Gene Transfection
Authors: Ueki Noboru , Ohkawa Toshihisa , Yokoyama Yuji , Maeda Juichiro , Kawai Yutaka , Ikeda Tatsuhiko , Amuro Yoshiki , Hada Toshikazu , Higashino Kazuya
Published: August 2005

This study was designed to assess whether the excessive secretion of transforming growth factor‐β1 (TGF‐β1) by Chinese hamster ovary (CHO) cells transfected with TGF‐β1 gene may be linked to the development of a metastatic phenotype. We observed large numbers of metastatic colonies in the lungs of nude mice inoculated with the transfected CHO cells. The tumors derived from these transfected cells demonstrated marked angiogenesis. We postulate that the overproduction of TGF‐β1 by these tumors may participate in the metastatic progression following establishment of angiogenesis at the primary tumor site.
Changes of Proliferative Activity and Phenotypes in Spontaneous Differentiation of a Colon Cancer Cell Line
Authors: Hara Ayumi , Hibi Toshifumi , Yoshioka Masahiro , Toda Kyoko , Watanabe Noriaki , Hayashi Atsushi , Iwao Yasushi , Saito Hidetsugu , Watanabe Tetsu , Tsuchiya Masaharu
Published: August 2005

We examined the alterations of proliferative activity and c‐myc expression of a colon cancer cell line (Caco‐2) during its spontaneous differentiation. Caco‐2 cells were cultured in various types of media and the degree of differentiation was monitored in terms of dome formation in cell monolayers and expression of alkaline phosphatase (ALP) activity. In Caco‐2 cells cultured with Eagle's minimum essential medium (EMEM) containing 10% fetal calf serum (FCS), dome formation was demonstrated and ALP activity was markedly increased after the cells reached confluence. Five‐fold reduction of c‐myc mRNA and a marked decrease in S‐phase cells were observed in the differentiated cells. These changes were not induced in FCS‐free EMEM. The addition of insulin and transferrin to FCS‐free EMEM did not induce cell differentiation or reduction of c‐myc mRNA expression. When Caco‐2 cells were cultured with three different serum‐free media, the induction of dome formation and the increase of ALP activity were observed to varying degrees. Expression of c‐myc mRNA in the cells cultured with one serum‐free medium decreased to a level similar to that in fully differentiated cells cultured with EMEM containing 10% FCS. These results suggest that a spontaneous switch from a proliferative state with high c‐myc expression to differentiated phenotype occurs after cells reach confluence and depends on the culture conditions.