bluuebunny/crossref_metadata_2025 huggingface dataset:
Mutational Analysis of CDKN2 (CDK4I/MTS1) Gene in Tissues and Cell Lines of Human Prostate Cancer
Authors: Komiya Akira , Suzuki Hiroyoshi , Aida Sara , Yatani Ryuichi , Shimazaki Jun
Published: August 2005

To study mutation of the CDKN2 gene in prostate cancer, samples from 51 Japanese patients and four human prostate cancer cell lines were examined by single‐strand conformation polymorphism analysis and direct sequencing. Only one out of 51 (2%) patients revealed a mutation, which was a 24 bp deletion from the 5′‐untranslated region to codon 3, resulting in loss of the initiation site. One of the four cell lines revealed a missense mutation, a GAC→TAC (Asp→Tyr) at codon 84. These results indicate that mutation of the CDKN2 gene is rare in prostate cancer and thus does not contribute significantly to the pathogenesis of human prostate cancer. Prostate cancer cell lines may acquire more frequent abnormality of the CDKN2 gene than tumor tissues.
Site‐specific Effects of Testosterone Propionate on the Prostate of Rat Pretreated with 3,2′‐Dimethyl‐4‐aminobiphenyl:Dose‐dependent Induction of Invasive Carcinomas
Authors: Shirai Tomoyuki , Tamano Seiko , Sano Masashi , Imaida Katsumi , Hagiwara Akihiro , Futakuchi Mitsuru , Takahashi Satoru , Hirose Masao
Published: August 2005

It has been shown that testosterone propionate (TP) strongly promotes induction of invasive carcinomas in previously initiated accessory sex organs. In this study, in order to clarify the dose‐dependence of this promotion, TP was given at 3 different levels (high, medium or low doses) using different sizes (2,1 and 0.5 cm. long) of Silastic tube for 40 weeks after administration of 3,2′‐dimethyl‐4‐aminobiphenyl to male F344 rats. The data showed development of invasive carcinomas in the dorso‐lateral and anterior prostate and in the seminal vesicle to be dose‐dependent with the high dose of TP being most effective for tumor induction. Average levels of serum testosterone were approximately 800, 600, 300 and 150 ng/dl in rats given the high to low doses and in control rats, respectively. Development of neoplastic lesions in the ventral prostate demonstrated an inverse dependence on the dose of TP. These findings, together with previous data, suggest that the tumor‐promoting potential of TP on rat prostate is unlikely to be simply due to its androgenic action and other factors should also be considered.
Changes in Radiation Sensitivity of Human Osteosarcoma Cells after p53 Introduction
Authors: Miyakoshi Junji , Yamagishi Nobuyuki , Ohtsu Shuji , Takebe Hiraku
Published: August 2005

Human osteosarcoma SAOS‐2 cells, which have a deletion in p53 gene, were transfected with plasmid pMSVneop53 containing human p53 cDNA and neomycin‐resistance gene. Three clones (SAOS‐MC10, SAOS‐MC11 and SAOS‐MC43) among 60 clones expressed p53 mRNA. No p53 protein was observed in SAOS‐MC10, while SAOS‐MC11 and SAOS‐MC43 produced p53 protein. The molecular weight of p53 protein in SAOS‐MC43 was lower than that in SAOS‐MC11. SAOS‐MC11 and SAOS‐MC43 were more sensitive and more resistant, respectively, to ionizing radiation than the parental SAOS‐2. We suggest that exogenous p53 protein might be one of the factors determining cellular radiosensitivity.
Apoptosis in Gastric Carcinomas and Its Association with Cell Proliferation and Differentiation
Authors: Saegusa Makoto , Takano Yasuo , Wakabayashi Tomo , Okayasu Isao
Published: August 2005

The significance of apoptosis in human gastric carcinomas was investigated in comparison with proliferative activity and p53 accumulation, using an in situ DNA nick end labeling method and immunohistochemistry for both Ki‐67 antigen and p53 protein. Apoptotic labeling indices (LI) of 51 differentiated carcinomas (21 of early and 22 of advanced stage) were significantly lower than for 33 undifferentiated tumors (9 of early and 24 of advanced stage) (P<0.05). In both types, apoptotic LI of advanced stage lesions were significantly higher than for the early stage cases (P<0.005, P<0.03). The distribution of apoptotic cells was different from that of Ki‐67‐positive cells, generally exhibiting an inverse correlation for areas of predominance. In contrast, there was no significant correlation between p53 immunoreactivity and either apoptotic LI or Ki‐67 LI. It is concluded that in human gastric carcinomas the susceptibility to apoptosis is related to tumor cell differentiation and depth of invasion, and may play a role in selection of clonal subpopulations with high growth potential.
Regional Mapping of Two Subunits of Transcription Factor E4TF1 to Human Chromosome
Authors: Sawada Jun‐ichi , Goto Masahide , Watanabe Hajime , Handa Hiroshi , Yoshida Michihiro C.
Published: August 2005

Genes of human transcription factor E4TF1 subunits, E4TF1a and E4TF1b, were mapped to region q21 of human chromosome 21 and to region q11.21 of human chromosome 7, respectively, by using a fluorescence in situ hybridization method.
Helicobacter pylori Infection, Serum Pepsinogen Level and Gastric Cancer: A Case‐Control Study in Japan
Authors: Fukuda Haruhiko , Saito Daizo , Hayashi Shuya , Hisai Hiroyuki , Ono Hiroyuki , Yoshida Shigeaki , Oguro Yanao , Noda Takeshi , Sato Toshiya , Katoh Masaru , Terada Masaaki , Sugimura Takashi
Published: August 2005

We conducted a case‐control study to evaluate the effect of Helicobacter pylori (HP) infection on the risk of gastric cancer in Tokyo, Japan. The sera at the time of diagnosis from 282 gastric cancer cases and 767 sex‐ and age‐matched cancer‐free controls were tested for the presence of anti‐HP IgG antibody (HM‐CAP ELISA kit) and serum pepsinogen (PG) level (PG I and PG II Riabead). No significant association was observed in all sets [matched odds ratio (OR)=1.04, 95% confidence interval: 0.73–1.49]. In subgroup analyses, however, an association was suggested in females [OR=1.57], a younger population (<50 years) [OR=1.86], early cancer [OR=1.53] and small cancer (<40 mm) [OR=1.55]. Furthermore, we observed a tendency for odds ratios to decrease with an increase in age or cancer growth (depth of tumor invasion and tumor size). Considering that the spontaneous disappearance of HP due to extended mucosal atrophy may lead to these decreasing odds ratios, we applied the conditional logistic model adjusted for the PG I/II ratio as a measure of atrophic gastritis. This analysis showed a positive association with HP infection in all sets [OR=1.69; 1.01–2.81], distal cancer [OR=1.88; 1.07–3.31] and intestinal‐type cancer [OR=3.76; 1.39–10.18]. We concluded that the risk of cancer associated with HP infection may be underestimated in studies with cross‐sectional exposure because of spontaneous disappearance of HP due to extended mucosal atrophy.
Establishment and Characterization of Acquired Resistance to Platinum Anticancer Drugs in Human Ovarian Carcinoma Cells
Authors: Misawa Toshiya , Kikkawa Fumitaka , Maeda Osamu , Obata Naoko H. , Higashide Koji , Suganuma Nobuhiko , Tomoda Yutaka
Published: August 2005

To investigate differences in resistance to cis‐diamminedichloroplatinum(II) (CDDP) and diammine (1,1‐cyclobutanecarboxylato)platinum(II) (CBDCA), and their newly developed derivative, ((—)‐(R)‐2‐aminomethylpyrrolidine (1,1‐cyclobutanedicarboxylato)platinum (II)) (DWA2114R), four types of resistant cell lines were established from a parental cell line (NOS2) of a serous cystadeno‐carcinoma of the ovary. The cross‐resistance of CDDP‐resistant cells (NOS2CR1 and NOS2CR2) to DWA2114R was slight (only 25% of the resistance to CDDP), and no cross‐resistance was observed to anticancer drugs other than the CDDP derivative, except to camptothecin (CPT‐11) in the case of NOS2CR2 cells. The cross‐resistance of CBDCA‐resistant cells (NOS2CBR) to DWA2114R was slight (only about 1/3 of the resistance to CBDCA), and no cross‐resistance was observed among anticancer drugs other than the CDDP derivative. On the other hand, DWA2114R‐resistant cells (NOS2DR) showed a high cross‐resistance to CDDP, CBDCA, etoposide (VP‐16), and CPT‐11. Intracellular accumulations of CDDP and CBDCA were markedly reduced in NOS2CR1, NOS2CR2, and NOS2CBR cells compared to those in NOS2 cells, but were reduced only slightly in NOS2DR cells. Intracellular accumulation of DWA2114R was reduced somewhat in the four types of resistant cells. Glutathione S‐transferase activity was not increased in any of the four types of resistant cells, and intracellular GSH concentration was increased only in NOS2CR2 cells (by 2.6 fold). From these results, we consider that the resistance mechanisms against CDDP and CDBCA are similar, and reduction of intracellular drug accumulations is a significant factor. Resistance to DWA2114R differed from resistance to CDDP and CBDCA in both cross‐resistance spectrum and resistance mechanism, indicating that reduction in intracellular drug accumulation is not the major resistance mechanism.
Pharmacological Correlation between Total Drug Concentration and Lactones of CPT‐11 and SN‐38 in Patients Treated with CPT‐11
Authors: Sasaki Yasutsuna , Yoshida Yasushi , Sudoh Kenichi , Hakusui Hideo , Fujii Hirofumi , Ohtsu Tomoko , Wakita Hisashi , Igarashi Tadahiko , Itoh Kuniaki
Published: August 2005

The pharmacokinetics of 7‐ethyl‐10‐[4‐(1‐piperidino)‐1‐piperidino]carbonyloxycamptothecin (CPT‐11) and its active metabolite, 7‐ethyl‐10‐hydroxycamptothecin (SN‐38), were examined to establish the pharmacokinetic variability of the active lactones of CPT‐11 and SN‐38 in comparison with that of the total (lactone and carboxylates) plasma CPT‐11 and SN‐38. Twelve patients with malignancies were entered in the study. All received 100 mg/m2 of CPT‐11 by intravenous drip infusion over 90 min. Blood was sampled at 10 time points in heparin‐containing syringes. Analysis by high‐performance liquid chromatography showed that the ratio of CPT‐11 lactone to total CPT‐11 concentration was highest (66%) just after the end of infusion and gradually decreased to 30% at 24 h. Almost 70% of SN‐38 lactone was detected after the end of infusion and this decreased to 50% within 24 h. The standard errors of percent lactone of CPT‐11 or SN‐38 to total drug concentration at each sampling point were less than 12%. The area under the concentration‐time curve (AUC) of total CPT‐11 and that of total SN‐38 were significantly correlated with the AUCs of the lactone CPT‐11 and those of lactone SN‐38, respectively. We conclude that, for practical purposes, monitoring of total CPT‐11 and SN‐38 has essentially the same clinical significance as monitoring of lactone CPT‐11 and SN‐38.
Sex Hormone‐dependent Renal Cell Carcinogenesis Induced by Ferric Nitrilotriacetate in Wistar Rats
Authors: Deguchi Jiro , Miyamoto Masaki , Okada Shigeru
Published: August 2005

Ferric nitrilotriacetate (Fe‐NTA), an iron chelate, induces necrosis of renal proximal convoluted tubules as a consequence of lipid peroxidation, and a high incidence of renal cell carcinoma (RCC) is also observed in rats and mice. The incidence of RCC and the extent of lipid peroxidation are greater in males than females. In the present study, the effects of castration or ovariectomy, and sex hormone treatment on Fe‐NTA‐induced renal carcinogenesis in rats were examined. Male and female Wistar rats were each divided into 5 groups. In group 1, rats were sham‐operated and treated intraperitoneally (i.p.) with nitrilotriacetate (NTA). In group 2, sham‐operated rats were treated with Fe‐NTA (5‐10 mg iron/kg/day, i.p.). Castrated or ovariectomized rats treated with Fe‐NTA served as group 3, Group 4 or 5 was treated in the same way as group 3, but in addition received either testosterone (group 4) or estradiol (group 5). NTA, Fe‐NTA or sex hormone treatments were initiated 4 weeks after the operation. NTA or Fe‐NTA treatments were conducted for 12 weeks, and sex hormones were administered for 10 months. After 10 months of treatment, all rats were autopsied and both kidneys were examined histopathologically. In NTA‐treated groups, there was no pathological change in the kidneys. In Fe‐NTA‐treated groups (groups 2‐5), testosterone treatment or ovariectomy increased the incidence of RCC, and estradiol treatment or castration decreased the incidence of RCC (male: sham operation, castration and testosterone treatment > castration > castration and estradiol treatment, female: ovariectomy and testosterone treatment > ovariectomy > sham operation, ovariectomy and estradiol treatment). These results indicate that sex differences observed in the incidence of RCC induced by Fe‐NTA are dependent upon sex hormones.
A Large‐scale, Hospital‐based Case‐Control Study of Risk Factors of Breast Cancer According to Menopausal Status
Authors: Hirose Kaoru , Tajima Kazuo , Hamajima Nobuyuki , Inoue Manami , Takezaki Toshiro , Kuroishi Tetsuo , Yoshida Minoru , Tokudome Shinkan
Published: August 2005

We conducted a large‐scale, hospital‐based case‐control study to evaluate differences and similarities in the risk factors of female breast cancer according to menopausal status. This study is based on a questionnaire survey on life style routinely obtained from outpatients who first visited the Aichi Cancer Center Hospital between January 1, 1988 and December 31, 1992. Among 36,944 outpatients, 1,186 women with breast cancer detected by histological examination were taken as the case group (607 premenopausal women and 445 postmenopausal women) and 23,163 women confirmed to be free of cancer were selected as the control group. New findings and reconfirmed factors of breast cancer were as follows. 1) The risk of at least one breast cancer history among subjects’ first‐degree relatives was relatively high among pre‐ as well as post‐menopausal women. 2) A protective effect of physical activity against breast cancer was observed among both pre‐ and post‐menopausal women. 3) Dietary control decreased the risk of premenopausal breast cancer. 4) Current smoking and drinking elevated the risk of breast cancer in premenopausal women. 5) Decreasing trends of breast cancer risk were associated with intake of bean curd, green‐yellow vegetables, potato or sweet potato, chicken and ham or sausage in premenopausal women, while in postmenopausal women a risk reduction was associated with a more frequent intake of boiled, broiled and/or raw fish (sashimi). Further study will be needed to clarify the age group‐ and/or birth cohort‐specific risk factors for breast cancer among the young generation in Japan.
Induction of DNA Recombination by Activated 3‐Amino‐1‐methyl‐5H‐pyrido[4,3‐b]indole
Authors: Hiramoto Kazuyuki , Kanamitsu Shin‐ichi , Negishi Kazuo , Ikeda Hideo , Hayatsu Hikoya
Published: August 2005

To investigate the genotoxic properties of a food‐derived carcinogen, 3‐amino‐1‐methyl‐5H‐pyrido‐[4,3‐b]indole (Trp‐P‐2), we have tested whether Trp‐P‐2 and its metabolically transformed products can induce DNA recombinations. Trp‐P‐2 is a strong mutagen and its activated form, the N‐hydroxylated derivative, Trp‐P‐2(NHOH), is known to form DNA adducts and cause DNA chain cleavage. Using a system in which phage λ undergoes recombination inside host Escherichia coli, we have found that Trp‐P‐2(NHOH), but not Trp‐P‐2 itself, can induce recombination. A nitroso derivative of Trp‐P‐2, Trp‐P‐2(NO), which can be reduced intracellularly to form Trp‐P‐2(NHOH), also induced recombination. Active oxygens are implicated in this recombinogenic action, since Trp‐P‐2(NHOH) is known to undergo spontaneous oxidative degradation, generating active oxygen radicals which can cause DNA chain cleavages. 4‐Hydroxyaminoquinoline N‐oxide and phenyl‐hydroxylamine also showed recombinogenic actions in this assay system; hence, it is suspected that aromatic amine‐type carcinogens have this property in common.
Preferential Production of Interleukin‐1β over Interleukin‐1 Receptor Antagonist Contributes to Proliferation and Suppression of Apoptosis in Leukemic Cells
Authors: Furukawa Yusuke , Kikuchi Jiro , Terui Yasuhito , Kitagawa Seiichi , Ohta Masatsugu , Miura Yasusada , Saito Masaki
Published: August 2005

Normal human monocytes were isolated in a nascent state by centrifugal elutriation and used for the study of interleukin‐1 (IL‐1) and interleukin‐1 receptor antagonist (IL‐1ra) expression. Neither IL‐1β nor IL‐1ra mRNA was present in monocytes just after the isolation, but they were induced simultaneously in response to various stimulants. In contrast, only IL‐1β mRNA was expressed in monocytic leukemia cell line JOSK‐1, while little or no IL‐1ra mRNA was detected even after stimulation. Dominant expression of IL‐1β over IL‐1ra was also observed in fresh leukemia cells including monocytic leukemias, i.e., IL‐1βmRNA was constitutively expressed in 26 out of 36 cases (72.2%), whereas IL‐1ra mRNA was present only in 8 cases (22.2%). The signal intensity of IL‐1βmRNA was stronger than that of IL‐1ra even in IL‐1ra‐positive cases. Apoptotic cell death of monocytes was significantly inhibited by IL‐1β, and it was enhanced by IL‐1ra. In fresh leukemia cells, 3H‐thymidine uptake was generally higher in IL‐1‐producing cases than in IL‐1ra‐producing cases, and was increased by the addition of IL‐1β in all cases tested. Cell proliferation was inhibited by either IL‐1ra or anti‐IL‐1β antibody in IL‐1‐producing cases, while it was enhanced by anti‐IL‐1ra antibody in IL‐1ra‐producing cases. These results suggest that the balance between IL‐1 and IL‐1ra is necessary for homeostasis of the mononuclear phagocytosis system. The imbalance between these two counter‐acting cytokines might contribute to the altered growth and accumulation of leukemic cells.
Immunoelectron Microscopic Localization of Gelatinase A in Human Gastrointestinal and Skin Carcinomas: Difference between Cancer Cells and Fibroblasts
Authors: Ohtani Haruo , Tabata Nobuko , Nagura Hiroshi
Published: August 2005

Previous reports revealed a discrepancy in gelatinase A localization in human cancers; i.e., protein localization in cancer cells and mRNA localization in stromal fibroblastic cells. To clarify this, we conducted immunoelectron microscopic study of gelatinase A in cancer and strmal cells in human gastrointestinal and skin carcinomas. Although both carcinoma cells and fibroblasts were positive for gelatinase A, the subcellular localizations were different. On immunoelectron microscopy, fibroblasts showed immunoreactivity in the lumen of the rough endoplasmic reticulum (rER) or in the cytosol on the surface of rER, demonstrating synthesis of the protein. Carcinoma cells showed diffuse deposition of gelatinase A in the cytosol, suggesting the accumulation of the antigen both in adenocarcinoma and squamous cell carcinoma. Immunoreactivity along the cell membrane was demonstrated in one case of skin carcinoma. Macrophages showed also diffuse deposition of gelatinase A in the cytosol. In conclusion, we found a qualitative difference of gelatinase A localization between carcinoma cells and fibroblasts, and concluded that carcinoma cells may not be important in the secretion of gelatinase A.
Loss of Heterozygosity on the Short Arm of Chromosome 9 without p16 Gene Mutation in Gastric Carcinomas
Authors: Sakata Ken , Tamura Gen , Maesawa Chihaya , Suzuki Yasushi , Terashima Masanori , Satoh Kunio , Eda Yoshiki , Suzuki Akihiko , Sekiyama Saburo , Satodate Ryoichi
Published: August 2005

A putative tumor suppressor gene, p16 (MST1; multiple tumor suppressor 1/CDK4I; cyclin‐dependent kinase 4 inhibitor), was isolated and mapped on the short arm of chromosome 9 (9p). The significance of p16 mutations in gastric tumorigenesis was examined by assessing p16 mutations as well as loss of heterozygosity (LOH) on 9p in 13 gastric adenomas and 45 adenocarcinomas. LOH on 9p (IFNA; α‐interferon locus) was detected in 22% (5/23 informative cases) of differentiated adenocarcinomas, 10% (1/10) of undifferentiated carcinomas and none (0/6) of the adenomas. Although we found a sequence polymorphism at the second position of codon 99 (CGC/CAC) of the p16 in one gastric adenoma patient, no somatic mutations were detected in any of the gastric adenomas or adenocarcinomas. These results suggest that p16 mutations probably do not contribute to gastric tumorigenesis. However, these data suggest that another tumor suppressor gene on 9p (near the IFNA locus) may contribute to the progression of differentiated adenocarcinoma of the stomach
Do Structural Changes of T Cell Receptor Complex Occur in Tumor‐bearing State?
Authors: Noda Satoshi , Nagata‐Narumiya Takako , Kosugi Atsushi , Narumiya Seiji , Ra Chisei , Fujiwara Hiromi , Hamaoka Toshiyuki
Published: August 2005

T cells in tumor‐bearing mice and cancer patients were recently shown to be devoid of CD3‐ζ chain, a signal‐transducing invariant chain in T cell receptor (TCR) complex, and p56lck tyrosine kinase. In the present study, we investigated the structure and function of TCR complex in T cells from BALB/c mice bearing CSA1M fibrosarcoma. The expressions of TCR chains and p56lck in a T cell‐enriched population from spleen were analyzed. Almost complete loss of CD3‐ζ and p56lck was observed in the preparation from tumor‐bearing mice as assessed by immunoblotting analysis using whole cell lysates, whereas the amounts of other TCR chains were relatively unchanged. However, these changes were due to the increase of contaminating Mac‐1+ cells in the spleen of tumor‐bearing mice because: 1) the removal of Mac‐1+ cells led to the restoration of CD3‐ζ and p56lck; and 2) CD3‐ζ was clearly present when the preparation was solubilized with ionic detergent. Fc receptor γ chain detected in the preparation from tumor‐bearing mice disappeared along with the removal of Mac‐1+ cells. These observations were further supported by the finding that addition of Mac‐1+ cells from tumor‐bearing mice to normal T cells resulted in loss of CD3‐ζ, leaving CD3‐ζ largely intact. When T cells from tumor‐bearing mice were highly purified by depletion of Mac‐1+ cells, these T cells contained normal amounts of CD3‐ζ at mRNA, protein, and surface levels, and expressed the properly assembled TCR complex on their cell surface. Moreover, stimulation of the TCR in these T cells by anti‐TCR antibodies resulted in a comparable Ca2+ mobilization to that observed in normal T cells. These results suggest that no structural changes occur in TCR complex in our tumor‐bearing mice, and that complete depletion of Mac‐1+ cells in important to assess the structure of TCR complex.
Nuclear Accumulation of p53 in Normal Human Fibroblasts Is Induced by Various Cellular Stresses which Evoke the Heat Shock Response, Independently of the Cell Cycle
Authors: Sugano Tatsuo , Nitta Makoto , Ohmori Hiroyuki , Yamaizumi Masaru
Published: August 2005

Nuclear accumulation of p53 is induced by various DNA damaging agents (the p53 response). Induction of nuclear accumulation of p53 after various cellular stresses, mostly other than DNA damage, including heat shock, was examined in normal human fibroblasts by immunostaining and flow cytometry using a mouse anti‐p53 monoclonal antibody. Immunostaining revealed nuclear accumulation of p53 within 6 h after various stresses [heat shock, osmotic shock, heavy metal (Cd), blockers of the cellular respiratory system (NaN3), amino acid analogues (azetidine and canavanine), an inhibitor of protein synthesis (puromycin), and oxygen free radicals (H2O2)]. Heat shock proved to be one of the most effective inducers among these stresses. FACScan analysis revealed that this induction of p53 occurred regardless of the stage in the cell cycle and that accumulation of cells in G2/M occurred. As all of these stresses are known to induce the heat shock response, the mechanism of p53 induction after stresses and that of heat shock response may share, at least partly, some common signaling pathway(s).
A novel SCID Mouse Model for Studying Spontaneous Metastasis of Human Lung Cancer to Human Tissue
Authors: Teraoka Seigo , Kyoizumi Seishi , Seyama Toshio , Yamakido Michio , Akiyama Mitoshi
Published: August 2005

We established a novel severe combined immunodeficient (SCID) mouse model for the study of human lung cancer metastasis to human lung. Implantation of both human fetal and adult lung tissue into mammary fat pads of SCID mice showed a 100% rate of engraftment, but only fetal lung implants revealed normal morphology of human lung tissue. Using these chimeric mice, we analyzed human lung cancer metastasis to both mouse and human lungs by subcutaneous inoculation of human squamous cell carcinoma and adenocarcinoma cell lines into the mice. In 60 to 70% of SCID mice injected with human‐lung squamous‐cell carcinoma, RERF‐LC‐AI, cancer cells were found to have metastasized to both mouse lungs and human fetal lung implants but not to human adult lung implants 80 days after cancer inoculation. Furthermore, human‐lung adenocarcinoma cells, RERF‐LC‐KJ, metastasized to the human lung implants within 90 days in about 40% of SCID mice, whereas there were no metastases to the lungs of the mice. These results demonstrate the potential of this model for the in vivo study of human lung cancer metastasis.
Synergistic Effect of Methionine‐depleting Total Parenteral Nutrition with 5‐Fluorouracil on Human Gastric Cancer: A Randomized, Prospective Clinical Trial
Authors: Goseki Narihide , Yamazaki Shigeru , Shimojyu Katsuo , Kando Fumio , Maruyama Michio , Endo Mitsuo , Koike Morio , Takahashi Hirokazu
Published: August 2005

Methionine‐depleting total parenteral nutrition (Met‐depleting TPN), infusing AO‐90 amino acid solution (lacking both L‐methionine and L‐cysteine) as a sole nitrogen source, showed synergistic effects with 5‐fluorouracil (5‐FU) in tumor‐bearing rats and in clinical trials with gastrointestinal tract cancers. In this study, the effect of Met‐depleting TPN with 5‐FU upon thymidylate synthase (TS) activity was examined, and the histological effect of this treatment on human gastric cancer was evaluated. Fourteen preoperative advanced gastric cancer patients were divided randomly into two groups. Seven cases were given Met‐depleting TPN for 7 days before surgery with continuous intravenous administration of 5‐FU (500 mg/body per day; total 4.0 g/body) (AO‐90 group). The other 7 received conventional L‐methionine‐containing TPN with 5‐FU (control group). All patients underwent gastrectomy without complications due to these treatments. Resected materials were examined for TS kinetics, and the anti‐cancer effect was also assessed histopathologically. The specimens in the AO‐90 group showed marked degeneration of cancer, while almost no effect was seen in the control group. The free TS activity of carcinoma tissue in the AO‐90 group was decreased and the TS inhibition rate was increased in comparison with the control group (P= 0.0165 and P= 0.0243, respectively). Met‐depleting TPN appears to play a role as a biomodulator of 5‐FU in human gastric cancer.
Enhancement of the Antitumor Effect by Combined Use of High‐energy Shock Waves and ATX‐70
Authors: Maruyama Michihiro , Asano Takehide , Uematsu Takeshi , Nakagohri Toshio , Hasegawa Masayuki , Miyauchi Hideaki , Iwashita Chikara , Isono Kaichi
Published: August 2005

The antitumor effects of high‐energy shock waves (HESW) in combination with ATX‐70 [a gallium‐porphyrin complex, 2,4‐bis(1‐decyloxyethyl)‐Ga(III)‐1,3,5,8‐tetramethylporphyrin‐6,7‐dipropionyi diaspartic acid] were investigated. In vitro, the cell damage to mouse MH134 hepatoma after HESW treatment was enhanced by adding ATX‐70. In vivo, HESW and ATX‐70 combination therapy inhibited cell growth. However, neither HESW treatment alone nor ATX‐70 treatment alone inhibited cell growth. These results imply that the antitumor effects of HESW and ATX‐70 combined therapy are caused by activation of ATX‐70 by HESW.
Inhibition of p53 Protein Phosphorylation by 9‐Hydroxyellipticine: A Possible Anticancer Mechanism
Authors: Ohashi Motoaki , Sugikawa Emiko , Nakanishi Noriyuki
Published: August 2005

Abnormality of p53, a tumor suppressor gene, is considered to be a potential cause of malignancy. We found that ellipticine and 9‐hydroxyellipticine (9HE), antitumor alkaloids, caused selective inhibition of p53 protein phospborylation in Lewis lung carcinoma and SW480 (human colon cancer cell line) in a concentration‐dependent manner from 0.1 to 100 μM. 9HE suppressed cdk2 kinase activity concentration‐dependently from 1 to 100 μM. By contrast, the inhibition of p53 protein phosphorylation by elliptinium and elliprabin (N2 substituted derivatives of 9HE) was very weak. A good correlation was observed between p53 phosphorylation inhibition and cytotoxic activity of these agents in terms of concentration‐response relationships, suggesting that inhibition of p53 protein phosphorylation via kinase inhibition may be involved in the anticancer mechanism of these agents. In addition, this study demonstrated that brief exposure to 9HE caused apoptosis of cancer cells. It is suggested that accumulation of dephosphorylated mutant p53 may induce apoptosis.
Characteristics of the Inhibitory Effect of Mitoxantrone and Pirarubicin on Lung Metastases of Colon Carcinoma 26
Authors: Iigo Masaaki , Shimamura Mariko , Sagawa Katsuhiko , Tsuda Hiroyuki
Published: August 2005

This study was performed to evaluate the antimetastatic activity of antitumor agents against metastatic colon carcinoma 26 (Co 26Lu), and to investigate their mechanisms of action. Pirarubicin demonstrated the most striking antitumor activity in mice bearing intravenously injected Co 26Lu cells. Etoposide and mitoxantrone also showed marked antitumor activity. Pirarubicin and mitoxantrone also exerted remarkable inhibitory effect on spontaneous lung metastases from subcutaneously implanted Co 26Lu. Pirarubicin showed marked inhibition of both primary tumor growth and lung metastases. Mitoxantrone was effective in preventing lung metastases even at doses that did not exhibit an antitumor effect on the primary tumor. Moreover, mitoxantrone administered two days after intravenous injection of tumor cells obviously reduced the number of lung colonies, while simultaneous injection of the drug did not inhibit colony formation. Mitoxantrone effectively inhibited angiogenesis on the chorioallantoic membrane at doses that did not affect the growth rate of embryos. These results suggest that mitoxantrone, besides its direct antitumor effect on tumor cells, may inhibit lung metastases by inhibiting angiogenesis.
The Positive Nuclear Staining Observed with Monoclonal Antibody against PRAD1/Cyclin D1 Correlates with mRNA Expression in Mantle Cell Lymphoma
Authors: Kuroda Hiroyuki , Komatsu Hirokazu , Nakamura Shigeo , Niitsu Yoshiro , Takahashi Toshitada , Ueda Ryuzo , Seto Masao
Published: August 2005

Recently, we produced a monoclonal antibody, 5D4, against the PRAD1/cyclin D1 product and suggested positive nuclear staining to be associated with mantle cell lymphoma (MCL). Now we have further characterized the specificity of this antibody and studied the relation of immunohistochemical detection to PRAD1/cyclin D1 mRNA expression and DNA rearrangement. Immunofluorescence and immunoblotting studies demonstrated the 5D4 antibody to be crossreactive with cyclin D2, but not cyclin D3. On immunostaining, 15 of 19 MCL cases (79%) presented the nuclear staining pattern and PRAD1/cyclin D1 mRNA expression was detected by Northern blot analysis in 12 of 15 MCL cases studied (80%); all cases with the mRNA expression showed the nuclear staining pattern. Southern blot analysis with 11q13 BCL‐1 probes detected DNA rearrangements in 8 of 19 MCL cases (42%), all 8 exhibiting PRAD1/cyclin D1 mRNA expression. In 21 lymphoma cases of types other than MCL, neither the mRNA expression nor the nuclear staining were observed, although cytoplasmic staining was often apparent. These results indicated that positive nuclear staining of lymphoma cells by 5D4 antibody reflects PRAD1/cyclin D1 mRNA expression, and showed that this monoclonal antibody has diagnostic value for differentiating MCL from other types of lymphomas.
Phase I Study of Paclitaxel by Three‐hour Infusion: Hypotension Just after Infusion Is One of the Major Dose‐limiting Toxicities
Authors: Tamura Tomohide , Sasaki Yasutsuna , Nishiwaki Yutaka , Saijo Nagahiro
Published: August 2005

The primary objectives of this study were to determine the maximum tolerated dose (MTD) of paclitaxel administered by 3‐h infusion to patients with solid tumors, and to characterize the pharmacokinetics of a 3‐h infusion in comparison with those of a 24‐h infusion. Twenty‐seven patients each received one of six levels of paclitaxel, 105, 135, 180, 210, 240 and 270 mg/m2, with premedication. Two patients given 240 mg/m2 and one patient given 270 mg/m2 unexpectedly had grade 3/4 hypotension just after finishing the paclitaxel infusion. Peripheral neuropathy was also dose‐limiting at 270 mg/m2. Although granulocytopenia was significantly less severe than with a 24‐h infusion, more than half of the patients experienced grade 4 toxicity at doses of 240 or 270 mg/m2. Severe hypersensitivity reactions (HSRs) were not observed. Pharmacokinetic studies using high performance liquid chromatography demonstrated proportionally greater increases in the peak plasma concentration and area under the curve, and decreases in clearance and volume of distribution with increasing dose, suggesting non‐linear pharmacokinetics of paclitaxel when given by 3‐h infusion. The MTD of paclitaxel given as a 3‐h infusion was determined to be 240 mg/m2 with dose‐limiting toxicities of granulocytopenia, peripheral neuropathy and hypotension. Hypotension just after infusion, induced by 3‐h infusion of paclitaxel, is a new observation which has not been reported previously. The recommended dose for phase II study is 210 mg/m2. Although hypotension was observed as an unexpected toxic effect, paclitaxel could be administered safely over 3 h with premedication and proper monitoring, resulting in reduced myelotoxicity and with no increase in the incidence of HSRs as compared with a 24‐h infusion.
Disturbance of the Cell Cycle with Colchicine Enhances the Growth Advantage of Diethylnitrosamine‐initiated Hepatocytes in Rats
Authors: Tsutsumi Masahiro , Ohashi Kazuo , Tsujiuchi Toshifumi , Kobayashi Eisaku , Kobitsu Kunihiko , Kitada Hiromichi , Majima Toshimitu , Okajima Eijiro , Endoh Takehiro , Hasegawa Kiyohiko , Mori Toshio , Konishi Yoichi
Published: August 2005

The effect of cell cycle disturbance due to colchicine on the induction of enzyme‐altered foci during liver regeneration in rats was studied. For initiation, diethylnitrosamine (DEN) at a dose of 10 mg/ kg was injected intraperitoneally and partial hepatectomy (PH) was performed 4 h thereafter. Colchicine at doses of 0, 0.1, 0.25 and 0.5 mg/kg was injected intraperitoneally 1 and 3 days after the initiation, followed by application of selection pressure consisting of 2‐acetylaminofluorene (AAF) and carbon tetrachloride (CCl4) administration. As end point lesions, γ–glutamyltransferase (GGT)‐positive enzyme‐altered foci were assayed at week 5. There was no significant effect of colchicine on numbers of foci. However, a significant, dose‐dependent increase in the area of GGT‐positive lesions in the groups treated with colchicine was observed. Bromodeoxyuridine labeling indices were higher in foci induced in colchicine‐treated rats than in the untreated rats. In a separate experiment, serum glutamic pyruvic transaminase was not increased significantly after DEN and colchicine treatment, and the mitotic index at 6 days after PH was increased in the liver of colchicine‐treated rats. These results suggest that the cell cycle disturbance induced by colchicine causes more pronounced selective growth of cells initiated by DEN and colchicine, and this experimental model may be useful for analyzing the mechanisms underlying that growth advantage and the effects of cell cycle abnormalities in liver carcinogenesis.
Histopathological Analysis of Invasive Bladder Carcinomas Induced by 3,2′‐Dimethyl‐4‐aminobiphenyl in Hamsters
Authors: Cui Lin , Hasegawa Ryohei , Ogawa Kumiko , Yamada Yasuyuki , Takahashi Satoru , Shirai Tomoyuki
Published: August 2005

Histopathological characteristics of urinary bladder tumors induced in Syrian golden hamsters by 3,2′‐dimethyl‐4‐aminobiphenyl (DMAB) were analyzed. DMAB was subcutaneously injected in corn oil at a concentration of 100 mg/kg once a week for 20 weeks and ethinyl estradiol (EE) was administered in the diet at a dose of 0.75 ppm throughout the experiment. A small group of animals was killed at week 20 and all survivors were killed at week 50. Urinary bladder carcinomas were induced in 14 of 18 hamsters (78%; 0.89/animal) in the DMAB+EE group and 11 of 17 (65%; 0.88/animal) in the DMAB alone group in males, and in 11 of 14 (79%; 0.79/animal) in the DMAB+EE group and 4 of 5 (80%; 0.80/animal) in the DMAB alone group in females examined between weeks 20 and 50. All were non‐papillary invasive transitional cell carcinomas partly demonstrating glandular and/or squamous differentiation, and most carcinomas developed in the bladder neck. Degree of invasion was clearly correlated with degree of morphological atypism in the transitional cell carcinomas, hut not with squamous or glandular differentiation. No sex difference or modifying effect of EE on DMAB urinary bladder carcinogenesis was evident. No bladder carcinomas were observed in non‐DMAB‐treated animals.
Cloning of Mouse DAN cDNA and Its Down‐regulation in Transformed Cells
Authors: Ozaki Toshinori , Ma Jun , Takenaga Keizo , Sakiyama Shigeru
Published: August 2005

Recently, we have demonstrated that DAN gene product exhibits a tumor‐suppressive activity in vitro. We report here the cloning and sequencing of a mouse DAN cDNA that contains the entire coding region. Sequence analysis revealed that mouse DAN cDNA is 1691 nucleotides in length and contains an open reading frame of 178 amino acids. The deduced mouse DAN protein sequence shows 96% and 93% identity with the counterparts isolated from rat 3Y1 fibroblasts and normal human lung, respectively. Genomic Southern blot hybridization indicated that DAN gene exists as a single copy in the mouse genome. The expression of DAN gene was suppressed in a variety of transformed NIH3T3 cells when compared with that in the parental NIH3T3 cells.
Establishment and Characterization of High‐ and Low‐lung‐metastatic Cell Lines Derived from Murine Colon Adenocarcinoma 26 Tumor Line
Authors: Sakata Keita , Kozaki Ken‐ichi , Iida Ken‐ichi , Tanaka Rie , Yamagata Sadako , Utsumi Kazuhiko R. , Saga Shinsuke , Shimizu Satoru , Matsuyama Mutsushi
Published: August 2005

We established and characterized high‐ (LuM1) and low‐lung‐metastatic (NM11) cell lines derived from murine colon adenocarcinoma 26 tumor line. LuM1 cell line was established as a clonal cell line from a cultured cell mixture derived from a lung‐metastatic nodule after 7 sequential subcutaneous transplantations of lung‐metastatic tumors in the abdominal wall of BALB/c mice. NM11 cell line was established from a cultured cell mixture derived from a subcutaneous transplant of murine colon adenocarcinoma 26 tumor cells. LuM1 cells showed marked spontaneous lung metastases, but NM11 cells rarely did. High invasive potential of LuM1 cells was revealed by in vitro invasion assay using Matrigel reconstituted membranes. Rapid retraction was observed in monolayers of human umbilical vein endothelial cells and bovine aortic endothelial cells when LuM1 cells were added on the monolayers. Gelatin zymography and immunochemical examinations with monoclonal antibodies against gelatinase B (Mr 95,000 type IV collagenase) showed secretion of large amounts of the gelatinase by LuM1 cells.
Evaluation of the Relative Cytotoxic Effects of Anticancer Agents in Serum‐supplemented versus Serum‐free Media Using a Tetrazolium Colorimetric Assay
Authors: Tsai Chun‐Ming , Perng Reury‐Perng , Chang Kuo‐Ting , Venzon David , Gazdar Adi F.
Published: August 2005

Most cell culture and in vitro drug sensitivity assays utilize serum‐supplemented media (SSM). However, fully defined serum‐free media (SFM) offer several advantages and are being used increasingly for initiation and maintenance of cell cultures. Because serum inhibits the in vitro cytotoxicity of certain antineoplastic agents, we investigated the inter‐relationships between medium type, cell proliferation and cytotoxic effect. Twenty‐four human lung cancer cell lines were tested with nine anticancer agents in both media types. A semi‐automated tetrazolium (MTT) colorimetric assay was used for assaying cell survival. Cell lines initiated and maintained in SFM preferentially proliferated in that medium type or proliferated equally well in both media types. In contrast, cell lines established in SSM varied considerably in their medium of preference. There appeared to be a direct correlation or trend between cell proliferative rate and cytotoxicity of all drugs with the possible exceptions of methotrexate and carmustine. In general, the cell lines were more sensitive to anticancer agents when they were exposed in the culture medium in which they preferentially proliferated. Therefore, to determine the influence of culture media on cytotoxicity, we analyzed the data only from lines that replicated equally efficiently in both media. After correction for cell proliferative rate, SSM had a negative effect on the cytotoxic action of some drugs (especially methotrexate, 5‐fluorouracil and, to a lesser extent, mitomycin‐C). Our results demonstrate that fully defined SFM may be suitable for initiating cell lines and for use in in vitro cytotoxicity assays for selection of individualized therapy or for screening of new anti‐neoplastic agents, and thus may increase the number of antineoplastic agents that can be tested satisfactorily.
Multiple Primary Cancers with Microsatellite Instability: Report of a Case
Authors: Ichikawa Daisuke , Takahashi Toshio , Hashimoto Naoya , Hoshima Masakazu , Kitamura Kazuya , Yamane Tetsuro , Yamaguchi Toshiharu , Abe Tatsuo , Inazawa Johji
Published: August 2005

We report here a patient who developed a variety of tumors both synchronously and metachronously over a 2‐year period. The involved organs were the uterus, ureter, and small and large intestines. The patient underwent open surgery 3 times and polypcctomies 6 times. Postoperative histopathologic analysis showed 2 adenomas and 8 carcinomas. Genetic analysis revealed microsatellite instabilities at the tested loci in all 10 tumors, indicating that replication errors played an essential role in the tumorigenesis. Early identification of microsatellite instability could be useful for predicting development of additional primary cancers.
Induction of OX40, a Receptor of gp34, on T Cells by Trans‐acting Transcriptional Activator, Tax, of Human T‐Cell Leukemia Virus Type I
Authors: Higashimura Norikazu , Takasawa Naruhiko , Tanaka Yuetsu , Nakamura Masataka , Sugamura Kazuo
Published: August 2005

gp34, which we had identified as a target molecule of the trans‐activation by Tax of human T‐cell leukemia virus type I (HTLV‐I), has been found to bind OX40, a member of the tumor necrosis factor receptor family, resulting in growth stimulation of activated T cells. We here demonstrate that not only gp34 (OX40L), but also OX40 can be transcriptionally activated by Tax. Three Tax‐producing human T‐cell lines carrying the HTLV‐I genome expressed OX40 on their surfaces. Furthermore, Tax‐induced transcriptional activation of OX40 was shown in Tax‐inducible JPX‐9 cells. These results demonstrate that both OX40 and its ligand (gp34) are constitutively expressed on the surfaces of Tax‐expressing T lymphocytes, suggesting that the OX40L/OX40 system contributes to growth stimulation of the virus‐infected T cells.
Genomic Organization of Human Papillomavirus Type 18 in Cervical Cancer Specimens
Authors: Kitagawa Kyoko , Yoshikawa Hiroyuki , Onda Takashi , Kawana Takashi , Taketani Yuji , Yoshikura Hiroshi , Iwamoto Aikichi
Published: August 2005

It has been reported that cervical cancers positive for human papillomavirus (HPV) 18 have a poorer prognosis than those with other HPV types. To gain a better understanding of the aggressive character of HPV 18‐positive cancers, we examined the difference in genomic organization between HPV 18 and HPV 16 harbored in cervical cancers. We amplified E1 and E2 genes from 9 HPV 18‐positive and 31 HPV 16‐positive cervical cancers by polymerase chain reaction (PCR). At least one of the two early genes was missing in 3 out of 9 HPV 18‐positive cancers, while both PCRs were positive in all 31 HPV 16‐positive cancers (P<0.05). We then analyzed the 9 HPV 18‐positive cancers by 15 contiguous polymerase chain reactions covering collectively the whole HPV 18 genome. In addition to the three with a deletion of the E1 or E2 gene, one had a deletion in the E5 and L2 genes and one had an insertion in the long control region. The frequent alterations in genomic organization, especially preferential deletion of the E1 or E2 gene, may be related to the more aggressive properties of HPV 18‐positive cervical cancers.
Decreased Levels of 2‐Amino‐3‐methylimidazo[4,5‐f]quinoline‐DNA Adducts in Rats Treated with β‐Carotene, α‐Tocopherol and Freeze‐dried Aloe
Authors: Uehara Nobuaki , Iwahori Yoshio , Asamoto Makoto , Baba‐Toriyama Hiroyasu , Iigo Masaaki , Ochiai Masako , Nagao Minako , Nakayama Masafumi , Degawa Masakuni , Matsumoto Kazuyuki , Hirono Iwao , Beppu Hidehiko , Fujita Keisuke , Tsuda Hiroyuki
Published: August 2005

To assess mechanisms of chemoprevention of hepatocarcinogenesis by trans‐β‐carotene (β‐C), DL‐α‐tocopherol (α‐T), and freeze‐dried whole leaves of Kidachi aloe (Aloe), formation of 2‐amino‐3‐methylimidazo[4,5‐f]quinoline (IQ)‐DNA adducts was measured by 32P‐post‐labeling analysis, and CYP1A1 and CYP1A2 protein levels were analyzed by ELISA. Group 1 rats were fed diet containing 0.02%β‐C, 1.5%α‐T or 30% Aloe over an 8‐day period, while group 2 was given basal diet alone. On day 7, all animals were subjected to two‐thirds partial hepatectomy (PH). Twelve hours after PH, they received a single dose of the carcinogenic food pyrolysate IQ (100 mg/kg) intragastrically, to initiate hepatocarcinogenesis. Rats were killed 6, 12, 24 and 48 h after IQ administration. The levels of adducts, expressed as relative adduct labeling values in rats treated with β‐C, α‐T and Aloe, were decreased as compared with the control group at hour 24 (36 h after PH), with a significant difference in the case of the β‐C group (46.4% of the control value). Similarly, all showed a tendency for decrease at hour 48. Furthermore, the levels of CYP1A2, known to be responsible for activation of IQ, showed a significant reduction at hour 24. It is concluded that β‐C, and possibly also α‐T and Aloe, have the potential to reduce IQ‐DNA adduct formation, presumably as a result of decreased formation of active metabolites. The results may explain, at least in part, the previously observed inhibitory effects of these compounds on induction of preneoplastic hepatocellular lesions.
Inverse Correlation between mRNA Expression of Plasminogen Activator Inhibitor‐2 and Lymph Node Metastasis in Human Breast Cancer
Authors: Ishikawa Noboru , Endo Yoshio , Sasaki Takuma
Published: August 2005

We examined mRNA expressions of urokinase‐type plasminogen activator (u‐PA), its specific receptor (u‐PR), and plasminogen activator inhibitors (PAI‐1 and PAI‐2) in 50 human breast cancers by the reverse transcriptase‐polymerase chain reaction method. The expressions of the genes are discussed in relation to the clinicopathological findings. In the overall population in breast cancers, a low level of PAI‐2 expression was significantly associated with lymph node involvement (P < 0.0001). The u‐PA, u‐PR, and PAI‐1 expressions tended to be at high levels in such metastatic cancers. Also, positive expression of u‐PA, u‐PR, and PAI‐1 was significantly correlated with negative expression of PAI‐2. These results indicate that PAI‐2 may play a critical role in the regulation of extracellular matrix degradation during tumor cell invasion and metastasis, and the expression of PAI‐2 may be useful as a marker to evaluate the prognosis of breast cancers.