bluuebunny/crossref_metadata_2025 huggingface dataset:
Modification by Analgesics of Lesion Development in the Urinary Tract and Various Other Organs of Rats Pretreated with Dihydroxy‐di‐N‐propylnitrosamine and Uracil
Authors: Shibata Masa‐Aki , Sano Masashi , Hagiwara Akihiro , Hasegawa Ryohei , Shirai Tomoyuki
Published: August 2005

Effects of the analgesics phenacetin, acetaminophen and antipyrine on lesion development in the urinary tract and other organs in male F344 rats were investigated. Animals were concurrently administered with 0.1% dihydroxy‐di‐N‐propylnitrosamine (DHPN) in drinking water and 3.0% uracil in the diet for 4 weeks and then, starting 1 week after the cessation of this treatment, received basal diet or diet containing phenacetin, acetaminophen or antipyrine for 35 weeks. The occurrences of renal cell tumors were increased in the groups given phenacetin or antipyrine, as compared with the DHPN+uracil alone controls. Antipyrine, but not the two other compounds, also enhanced development of hyperplastic lesions in the renal pelvis and ureter. In the urinary bladder, phenacetin and antipyrine treatments were both associated with increased incidence of preneoplastic or neoplastic lesions. Furthermore, phenacetin alone, without the initiating agent pretreatments, induced simple hyperplasias of the urinary bladder at high incidence. Antipyrine enhanced induction of hyperplastic lesions in the ureter and was also found to increase the incidences of preneoplastic and neoplastic lesions in the liver. Although decreased incidences of tumor development of lung and thyroid were observed for the group given phenacetin, this might have been linked to the decreased weight gain. The results confirmed that combination treatment with DHPN+uracil is effective for wide‐spectrum initiation of carcinogenesis in the urological tract and demonstrated significant modification potential for both phenacetin and antipyrine.
Isolation and Properties of Tumor‐derived Endothelial Cells from Rat KMT‐17 Fibrosarcoma
Authors: Utoguchi Naoki , Dantakean Adul , Makimoto Hiroo , Wakai Yukiko , Tsutsumi Yasuo , Nakagawa Shinsaku , Mayumi Tadanori
Published: August 2005

Rat KMT‐17 fibrosarcoma‐derived endothelial cells were isolated by Percoll gradient centrifugation with an attaching‐speed separation technique, and their properties in culture were examined. The primary cultured tumor‐derived endothelial cells (TEC) showed angiotensin‐converting enzyme activity, positivity for Factor VIII‐related antigen staining, and typical capillary‐like formation on Matrigel. The primary cultured TEC monolayer showed greater permeability than normal tissuederived endothelial cell (aorta, vena cava and epididymal fat capillary) monolayers on FITC‐dextran diffusion (molecular weight 70,000). Leukocyte adhesion to TEC was reduced compared to that to fat‐derived capillary endothelial cells. These characteristics resembled those of tumor vascular endothelium, and were observed both in the primary and first‐passage cell cultures, but not in the fourth‐passage cell cultures. Our findings indicate that primary or subcultured TEC are applicable for studies of the physiological characteristics of tumor endothelial cells.
Acute Induction of Adriamycin‐resistance in Human Colon Carcinoma HT‐29 Cells Exposed to a Sublethal Dose of Adriamycin
Authors: Tomida Akihiro , Naito Mikihiko , Tsuruo Takashi
Published: August 2005

To study the mechanisms of the acute induction of drug resistance in cancer cells, we have established a model system in which adriamycin (ADM) induces immediate drug resistance. In this system, human colon carcinoma HT‐29 cells were pretreated for 1 h with a subtoxic dose of ADM (0.3 μg/ml) and incubated for 24 h in drug‐free medium. Then the cells were treated for 1 h with ADM, and the cell survival was determined in terms of colony‐forming ability. The survival of the pretreated cells was increased up to 100‐fold, as compared with that of untreated cells. Such increased survival, however, was observed only after high doses of ADM (2 to 8 μg/ml); more than 99% of the cells were killed. These results indicate that only a small fraction of ADM‐pretreated cells acquire the ADM‐resistant phenotype. Similar induced resistance was observed in five of seven subclones isolated from HT‐29 cells by limiting dilution, suggesting that the majority of cells in the parental HT‐29 population could acquire the ADM‐resistant phenotype. In the subclone HT‐29T9, the ADM pretreatment induced concomitant resistance to daunomycin, VP‐16, and VM‐26 but not to agents other than topoisomerase II inhibitors. The ADM‐induced drug resistance did not accompany MDR1 gene expression and could not be overcome by verapamil, a P‐glycoprotein inhibitor. The present system could be useful to study the acute induction mechanism(s) of ADM‐resistance, which could be relevant to clinical resistance in patients.
Infrequent Somatic Mutation of the MTS1 Gene in Primary Bladder Carcinomas
Authors: Kai Mikio , Arakawa Hirofumi , Sugimoto Yoshihisa , Murata Yasushi , Ogawa Michio , Nakamura Yusuke
Published: August 2005

We examined a candidate tumor suppressor gene on chromosome 9p21, MTS1/CDK4I (multiple tumor suppressor 1/cyclin‐dependent kinase 4 inhibitor), which has been found to be mutated frequently in cell lines derived from bladder carcinomas, for somatic mutations in 39 primary bladder cancers by means of SSCP (single‐stranded conformational polymorphism) and DNA sequencing. Mutations were detected in two of these carcinomas; one was a 61‐base deletion and the other a 1‐base deletion. In both cases the homologous allele was missing, indicating that “two‐hit”mutation of the MTS1 gene had taken place in these tumors. The results indicated that inactivation of the MTS1 gene is likely to be a contributing factor in some, but not the majority of, bladder cancers.
Increased Susceptibility to N‐Nitrosomethylurea Gastric Carcinogenesis in Transforming Growth Factor α Transgenic Mice with Gastric Hyperplasia
Authors: Tamano Seiko , Jakubczak John , Takagi Hitoshi , Merlino Glenn , Ward Jerrold M.
Published: August 2005

Glandular stomach carcinogenesis after N‐nitrosomethylurea (NMU) treatment was examined in transgenic mice bearing a human transforming growth factor alpha (TGF‐α) cDNA driven by the mouse metallothionein‐I promoter (mouse line MT100) in the inbred mouse line FVB/N. Untreated MT100 mice exhibit a severe age‐related gastric fundic hyperplasia. Both sexes of MT100 mice were given 10 weekly intragastric intubations of 0.5 mg NMU per mouse from 6 weeks of age and/or zinc chloride in drinking water to stimulate transgene expression from 5.5 weeks of age to the experiment termination. Animals were killed sequentially at 10, 19 and 29 experimental weeks. Several histochemical markers (AB‐PAS, TGF‐α, pepsinogen isozyme 1, proliferating cell nuclear antigen) were used. Abnormal histochemical patterns were found in untreated MT100 and NMU‐treated MT100 mice for all 4 markers of differentiation and carcinogenesis. Precancerous lesions including atypical and/or adenomatous hyperplasia were found in the fundic region of 16/22 male and 8/22 female MT100 mice but not in 27 male and 24 female FVB/N mice treated with NMU. One of 22 MT100 males had fundic carcinoma. FVB/N mice treated with NMU had neither precancerous lesions nor carcinomas in the fundus. Well differentiated adenocarcinomas in the pyloric region were induced at incidences of 2/22 male and 1/22 female MT100 mice treated with NMU and 4/27 male and 4/24 female FVB/N mice treated with NMU. Both strains also had a high incidence (55 to 92%) of squamous cell carcinomas of the forestomach. In conclusion, TGF‐α induced a hyperplastic lesion in the gastric fundus that appeared to predispose the MT100 mice to carcinogenesis by NMU.
Granulocyte‐Macrophage Colony‐stimulating Factor Augments Lymphokine‐activated Killer Activity from Pleural Cavity Mononuclear Cells of Lung Cancer Patients without Malignant Effusion
Authors: Takahashi Keiji , Sone Saburo , Saito Seiya , Kamamura Yoshitaka , Uyama Tadashi , Ogura Takeshi , Monden Yasumasa
Published: August 2005

The role of recombinant granulocyte‐macrophage colony‐stimulating factor (GM‐CSF) in augmentation of lymphokine‐activated killer (LAK) cell induction by interleukin‐2 (IL‐2) from pleural cavity mononuclear cells (PCMNCs) was examined in sixteen patients with resectable primary lung cancer not associated with malignant effusion. None of the patients had received any anticancer therapy prior to this study. Incubation of PCMNCs of patients without malignant effusion with GM‐CSF for 4 days in the presence of IL‐2 resulted in a significant increase in LAK activity against natural killer‐resistant Daudi cells. This result was obtained by using the 4 h 51Cr‐release assay. PCMNCs and blood mononuclear cells (BMNCs) were harvested simultaneously from pleural cavity lavage fluid and peripheral blood in lung cancer patients. The LAK activity developed from PCMNCs and BMNCs following incubation with IL‐2 for 4 days, but the LAK activity from PCMNCs was significantly lower than that from BMNCs (P < 0.05). Incubation of PCMNCs with GM‐CSF augmented the LAK activity from PCMNCs to a level as high as that from BMNCs. These results suggest that the combined use of GM‐CSF with IL‐2 may result in augmentation of LAK activity developed from PCMNCs of lung cancer patients without malignant effusion.
c‐Cbl Is Inducibly Tyrosine‐phosphorylated by Epidermal Growth Factor Stimulation in Fibroblasts, and Constitutively Tyrosine‐phosphorylated and Associated with v‐Src in v‐src‐transformed Fibroblasts
Authors: Odai Hideharu , Sasaki Ko , Hanazono Yutaka , Ueno Hiroo , Tanaka Tomoyuki , Miyagawa Kiyoshi , Mitani Kinuko , Yazaki Yoshio , Hirai Hisamaru
Published: August 2005

The c‐cbl gene was cloned as the cellular homolog of the v‐cbl oncogene that is the transforming component of a marine tumorigenic retrovirus, CAS NS‐1, though the biological roles of c‐Cbl remain to be elucidated. We have previously reported that c‐Cbl is implicated in the signal transduction triggered by granulocyte‐macrophage colony‐stimulating factor or erythropoietin in hematopoietic cells. Here, we observed tyrosine phosphorylation of c‐Cbl in cells expressing epidermal growth factor receptor depending on EGF stimulation and in v‐src transformed cells. Furthermore, c‐Cbl was revealed to associate with v‐Src in vivo. By means of binding experiments using glutathione S‐transferase fusion proteins, we have found that the SH2 and SH3 domains of many proteins bind to c‐Cbl. These findings strongly suggest that c‐Cbl is implicated in a wide variety of signal transduction pathways, including those of EGF receptor and Src protein, as well as in the signaling pathways of hematopoietic cells.
Sensitivity and Specificity of Mass Screening for Gastric Cancer Using the Measurement of Serum Pepsinogens
Authors: Hattori Yasushi , Tashiro Hiromi , Kawamoto Toshihiro , Kodama Yasushi
Published: August 2005

The aim of this study was to estimate the validity of mass screening for gastric cancer using serum pepsinogens (PG test). The study subjects were 4876 workers aged from 40 to 61 years old. Sera were obtained at the time of the health checkup and serum pepsinogen levels (PG I and PG II) were measured at the same time. PG I < 50 ng/ml and PG I/PG II ratio < 3.0 were adopted as the criteria for a positive result for the PG test. PG test‐positive subjects were examined, in principle, by endoscopy. Furthermore, all the subjects were followed up for a year to check for occurrence of gastric cancer. Among the total subjects, 911 (18.7%) were positive for the PG test and 650 (71.4%) among the positive subjects underwent further examinations, which revealed 11 cases of gastric cancer. Seven gastric cancer cases were diagnosed during the follow‐up period within one year after the PG test. Six out of the 7 cancer cases had been negative in the PG test. When the results of one year's follow‐up from the time of screening were defined as the gold standard, the sensitivity and specificity of the PG test were estimated at 66.7% and 81.5%, respectively. The authors conclude that the validity of the PG test as a mass screening method may be comparable to that of X‐ray screening, if optimum criteria of a positive test are selected.
Human T‐Lymphotropic Virus Type II in Japan
Authors: Yoshizaki Hitomi , Kitamura Katsuhiko , Oki Atsuko , Yahara Seiji , Yamanaka Retsuji , Fukushima Yoshiko , Komuro Katsutoshi , Yamazaki Shudo , Honda Mitsuo
Published: August 2005

Serum specimens were assayed for human T‐lymphotropic virus type II (HTLV‐II) infection in 1,500 individuals known to be seropositive for HTLV‐I and 30,000 blood donors in Japan. All HTLV‐I‐positive individuals were negative for HTLV‐II. However, one of the blood donors was clearly seropositive for HTLV‐II. Further, the donor was shown to be positive for HTLV‐IIb. Here we report at least one case with HTLV‐II in Japan and discuss the origin of the infection.
Trends in the Incidence of Female Breast and Cervical Cancers in Miyagi Prefecture, Japan, 1959–1987
Authors: Minami Yuko , Takano Akira , Okuno Yoshi , Fukao Akira , Kurihara Minoru , Hisamichi Shigeru
Published: August 2005

Trends in the incidence of female breast and cervical cancers were examined, using the cancer registry data in Miyagi Prefecture, Japan, during 1959–1987. The age‐standardized incidence rate of breast cancer has been increasing, while that of cervical cancer has been decreasing. Age‐period‐cohort models were applied to clarify the trends in incidence. For breast cancer incidence, the age‐period model adequately represented the data, and demonstrated that the risk of developing breast cancer has been increasing in recent time periods. The effect of cohort on breast cancer incidence was insignificant and the full model containing age, period and cohort showed irregularities in the cohort effect. For cervical cancer incidence, the effect of period was significant, while the effect of cohort was marginal. The full model containing age, period and cohort showed that cervical cancer risk has been decreasing in recent time periods and younger birth cohorts. Using published reports, we investigated the trends in the prevalence of various risk factors and compared them with the trends in the incidence at both sites. It is suggested that the effects of period and cohort might be related to the changes in the prevalence of these risk factors as well as to improvements of the diagnostic procedures.
Influence of Paternal 252Cf Neutron Exposure on Abnormal Sperm, Embryonal Lethality, and Liver Tumorigenesis in the F1 Offspring of Mice
Authors: Watanabe Hiromitsu , Takahashi Tadateru , Lee Juing‐Yi , Ohtaki Megu , Roy Goutam , Ando Yasumi , Yamada Kazumasa , Gotoh Takahiko , Kurisu Kazunobu , Fujimoto Nariaki , Satow Yukio , Ito Akihiro
Published: August 2005

Experiments were conducted to determine whether neutron‐induced genetic damage in parental germline cells can lead to the development of cancer in the offspring. Seven‐week‐old C3H male mice were irradiated with 252Cf neutrons at a dose of 0, 50, 100, or 200 cGy. Two weeks or 3 months after irradiation, the male mice were mated with virgin 9‐week‐old C57BL females. Two weeks after irradiation, the irradiated male mice showed an increased incidence of sperm abnormalities, which led to embryo lethalities in a dose‐dependent manner when they were mated with unirradiated female mice. Furthermore, liver tumors in male offspring of male mice in the 50 cGy group were significantly increased in 19 of 44 (43.2%) animals, in clear contrast to the unirradiated group (1 of 31; 3.2%) (P < 0.01). In the 100 cGy group, 6 of 39 (15%) mice had lesions. At 3 months after irradiation abnormal sperm and embryonal lethality were not significantly increased. The incidences of liver tumors in male offspring from the 50 cGy, 100 cGy and 200 cGy groups were 6 of 20 (30%), 5 of 22 (23%) and 1 of 19 (5%), respectively, which are not significantly increased compared with the control. It is concluded that increased hepatic tumor risk in the F1 generation may be caused by genetic transmission of hepatoma‐associated trait(s) induced by 252Cf neutron irradiation.
Establishment of an Undifferentiated Leukemia Cell Line (Kasumi‐3) with t(3;7)(q27;q22) and Activation of the EVI1 Gene
Authors: Asou Hiroya , Suzukawa Kazumi , Kita Kenkichi , Nakase Kazunori , Ueda Haruo , Morishita Kazuhiro , Kamada Nanao
Published: August 2005

A novel human leukemia cell line (Kasumi‐3) was established from the blast cells of a 57‐year‐old man suffering from myeloperoxidase‐negative acute leukemia. The cell line had five distinctive features, as follows. 1) Flow cytometric analyses showed cell surface expression of CD7, CD4, CD13, CD33, CD34, HLA‐DR and c‐Kit. This phenotype is compatible with that of acute myelocytic leukemia cells with the M0 subtype in the French‐American‐British classification. 2) Kasumi‐3 cells carried chromosomal abnormalities of t(3;7)(q27:q22), del(5)(q15), del(9)(q32), and add(12)(pll). The breakpoint of 3q27 was located near the EVI1 gene, and a high level of expression of the EVI1 gene was observed. 4) Kasumi‐3 cells treated with TPA showed maturation to monocytic lineage. 5) Treatment with either interleukin (IL)‐2, IL‐3, IL‐4, granulocyte‐macrophage colony‐stimulating or stem cell factor induced the proliferation of Kasnmi‐3 cells. Thus, the Kasumi‐3 cell line shows the characteristic features of undifferentiated leukemia. It should, therefore, be useful both for studying the biological characteristics of acute myelogenous leukemia M0 subtype and for investigating the role of the EVI1 gene in leukemogenesis.
Flat Serrated Adenomas of the Colorectal Mucosa
Authors: Rubio Carlos A. , Jaramillo Edgar
Published: August 2005

A total of 47 flat serrated neoplasias of the colorectal mucosa are presented: 44 were flat serrated adenomas and the remaining 3 flat serrated adenocarcinomas arising in flat serrated adenomas. These lesions were found among 600 flat mucosal lesions removed at colonoscopy during a 3‐year period (1992 and 1994) at the Karolinska Hospital. Thirty‐five of the 47 patients (74%) were males and the remaining 12 (26%), females. Depending upon the degree of cellular dysplasia within the epithelium, serrated adenomas were divided into those with low‐grade dysplasia (LGD), when the dysplastic nuclei were present in the deeper half of the epithelium, and those with high‐grade dysplasia (HGD), when the dysplastic nuclei were found even in the upper half of the epithelium. LGD was present in 37 (84.1%) of the 44 serrated adenomas and HGD in the remaining 7 (15.9%). Depending upon the topographic distribution of the dysplastic epithelium within the crypts, flat serrated adenomas were divided into type I, when the dysplastic epithelium was limited to the lower half of the serrated crypts, and type II, when the dysplastic epithelium was even present in the superficial half of the serrated crypts. Of the 44 serrated adenomas, 38 (86.1%) were type I and the remaining 6 (13.9%) type II. The dysplastic epithelium seemed to originate at the base of the crypts and to progress upwards, replacing the scalloped, serrated epithelium of the sides of the crypts. Invasive adenocarcinomas (i.e., with submucosal extension) were seen to arise from flat serrated adenomas with LGD type I (n=2) or with HGD type II (n= 1). This preliminary survey suggests that flat serrated adenomas of the colorectal mucosa may be lesions with a propensity to evolve into invasive adenocarcinoma, irrespective of the degree of the epithelial dysplasia or of their extension along the crypts.
Antimetastatic Effect of a Novel Indolocarbazole (NB‐506) on IMC‐HM Murine Tumor Cells Metastasized to the Liver
Authors: Arakawa Hiroharu , Matsumoto Hiroyoshi , Morita Masashi , Sasaki Minoru , Taguchi Kazuhiro , Okura Akira , Nishimura Susumu
Published: August 2005

IMC‐HM cells were isolated from spontaneously induced ascitic IMC carcinoma cells that had been maintained intraperitoneally in CDF1 mice. Metastasis to the liver of subcutaneously implanted IMC‐HM cells was detected 10 days after implantation into the flanks of mice (day 10), but metastasis to other organs was limited. Thereafter, however, tumor cells spread rapidly to lymph nodes, lung, spleen, ovary and other organs, and the mice died on day 13 to 18. We report here, together with the properties of IMC‐HM cells, the effects of adriamycin, cisplatin, etoposide and a new indolocarbazole antitumor compound (NB‐506) on this model of metastasis. Although these anticancer agents all inhibited the growth of the subcutaneous tumors, their effects on the life span of the tumor‐bearing mice varied. Treatment with NB‐506, started on day 1, more than doubled the survival period at doses 30 mg/m2 to 900 mg/m2. Further, treatment with NB‐506, started on day 4 after resection of the primary tumor, inhibited growth of the metastasized tumor in the liver and other organs. Etoposide also increased the life span at a limited range of doses. However, the life‐prolonging effects of adriamycin and cisplatin were marginal. These results demonstrate that IMC‐HM carcinoma is a good model for spontaneous metastasis to the liver followed by lethal spread to many organs. Moreover, NB‐506 was found to be highly effective against the growth not only of subcutaneous tumors, but also of tumors metastasized to the liver.
Effects of Sex Steroids and Growth Factors on Migration and Invasion of Endoraetrial Adenocarcinoma SNG‐M Cells in vitro
Authors: Ueda Masatsugu , Fujii Hideji , Yoshizawa Keiko , Abe Fuminori , Ueki Minoru
Published: August 2005

Biological effects of sex steroids (estradiol‐17β, E2; progesterone, P; medroxyprogesterone acetate, MPA; Danazol, DZ) and growth factors (epidermal growth factor, EGF; transforming growth factor, TGF‐α, β) on migration and invasion of endometrial adenocarcinoma SNG‐M cells were investigated by haptotactic migration and haptoinvasion assay. The enzymatic degradation of the extracellular matrix hy tumor cells was also examined. Tumor cell migration along a gradient of substratum‐bound fibronectin was inhibited by 0.1–10 μM MPA and DZ, but promoted by 0.1–10 nM EGF and TGF‐a in a concentration‐dependent manner. E2, P and TGF‐β did not have any effect on the motility of tumor cells. These effects were also confirmed by wound assay. The invasive activity of SNG‐M cells into reconstituted basement membrane (Matrigel) was inhibited by the presence of 0.1–10 μM MPA and DZ, but promoted by 0.1–10 nM EGF and TGF‐α in a concentration‐dependent manner. E2, P and TGF‐β did not have any effect on tumor cell invasion. The zymography of tumor‐conditioned medium showed that the treatment of SNG‐M cells with EGF and TGF‐α resulted in the increase of the 68, 72 and 92 kDa type IV collagenases (matrix metalloproteinase, MMP‐2 and 9). Sex steroids and TGF‐j9 did not have significant effects on MMP‐2 and 9. Stromelysin (MMP‐3), also secreted hy SNG‐M cells, was not affected by sex steroids and growth factors. These results suggest that EGF and TGF‐a act as positive regulators on the invasion process of endometrial adenocarcinoma cells, which may partly he associated with the induction of type IV collagenase secretion by tumor cells. The inhibitory effects of MPA and DZ on tumor cell invasion may depend at least partly on their inhibitory action on the motility of tumor cells.
Tobacco, Alcohol and Dietary Factors Associated with the Risk of Oral Cancer among Japanese
Authors: Takezaki Toshiro , Hirose Kaoru , Inoue Manami , Hamajima Nobuyuki , Kuroishi Tetsuo , Nakamura Shigeo , Koshikawa Takashi , Matsuura Hidehiro , Tajima Kazuo
Published: August 2005

The mortality rate among Japanese men due to oral cancer is increasing, but risk factors among Japanese other than smoking and drinking have not been examined. To investigate the dietary factors involved in oral cancer, we conducted a hospital‐based case‐referent study in Aichi, Japan. Cases comprised 189 men and 77 women aged 20–79 years with one of the following cancers: tongue, mouth, oropharynx and hypopharynx. The reference group comprised 9,858 male and 26,669 female outpatients without cancer. Smoking and drinking were highly associated with an increased risk of oral cancer. Japanese sake showed a lower odds ratio (OR) than beer or hard liquor (OR=3.6, 4.5 and 4.8, respectively). In the cross analysis between smoking and drinking, smoking combined with drinking increased the risk of oral cancer to three times that of smoking only (OR=6.2 vs. 2.2). Frequent intake of raw vegetables (OR=0.5) and fruit (OR=0.5) were inversely associated with the risk of oral cancer after adjustment for age, sex, smoking, drinking and year of visit. Western‐style breakfast and salty food preference decreased the risk of oral cancer, and salty food preference was still statistically significant by multivariate analysis (OR=0.7). In conclusion, smoking cessation, drinking control and frequent intake of raw vegetables and fruit among Japanese are likely to be effective preventive measures against oral cancer.
Direct Effects of Testosterone, Dihydrotestosterone and Estrogen on 3,2′‐Dimethyl‐4‐aminobiphenyl‐induced Prostate Carcinogenesis in Castrated F344 Rats
Authors: Mori Toshio , Cui Lin , Kato Koji , Takahashi Satoru , Imaida Katsumi , Iwasaki Shogo , Ito Nobuyuki , Shirai Tomoyuki
Published: August 2005

The present experiment was carried out to explore the effect of endogenous androgen on rat prostate carcinogenesis induced by 3,2′‐dimethyl‐4‐aminobiphenyl (DMAB) and testosterone propionate (TP) or 5α‐dihydrotestosterone (DHT) with or without ethinyl estradiol (EE). In order to eliminate the influence of endogenous androgen, F344 rats were orchiectomized just after initiation with the prostate carcinogen, DMAB, and then given TP, DHT, TP plus EE or DHT plus EE for 40 weeks. The results demonstrated that while administration of TP following DMAB treatment causes invasive carcinomas in the lateral and anterior prostate and seminal vesicles, DHT does not exhibit equivalent effects. Synergistic enhancement was also evident with TP plus EE, but not with DHT plus EE. The incidences of prostatic and seminal vesicle lesions in all groups of the present experiment, except for the group given castration without hormonal supplement, were equivalent to those previously found in non‐castrated animals. Therefore, the present findings indicate that endogenous testosterone may not be required for promotion hy TP/EE of DMAB‐initiated prostate carcinogenesis and that it may not contribute to the actions of DHT.
Early Alterations of Apoptosis and Cell Proliferation in Azoxymethane‐initiated Rat Colonic Epithelium
Authors: Hirose Yoshinobu , Yoshimi Naoki , Makita Hiroki , Kara Akira , Tanaka Takuji , Mori Hideki
Published: August 2005

Alterations of apoptosis and cell proliferation in the colonic epithelium of rats after exposure to azoxymethane (AOM) were estimated by means of the terminal deoxynucleotidyl transferasemediated dUTP‐biotin nick end labeling (TUNEL) method, measurement of 5‐bromo‐2′‐deoxyuridine (BrdU) incorporation, immunohistochemical staining for proliferating cell nuclear antigen (PCNA), and counting of raitotic cells. F344 male rats were given a single s.c. injection of AOM (15 mg/kg body weight) at 6 week of age, and killed 4 h, 8 h, 3 days, and 7 days after the AOM treatment. At 4 h after the treatment, many damaged cells were already observed in the colonic epithelium, and they were positive by TUNEL staining. At 8 h, the number of TUNEL‐positive cells was largest. The reduction of DNA synthesis in the colonic epithelium, confirmed by BrdU incorporation, was not distinct in comparison with the mitotic inhibition. There was no remarkable change in PCNA labeling index, except that strong expression of PCNA was detected in many damaged cells. On the 3rd day, the appearance of cell death became infrequent and an increase of cell proliferation occurred. On the 7th day, the expression of TUNEL and the cell proliferation biomarkers were at almost normal levels. These findings suggest that AOM induces apoptosis, which is associated with synchronous inhibition of mitosis. The data also indicate that PCNA immunostaining does not reflect the true proliferation state in the early phase after AOM exposure, probably due to the occurrence of cell cycle arrest or DNA repair.
Lack of Chemoprevention Effects of the Monoterpene d‐Limonene in a Rat Multi‐organ Carcinogenesis Model
Authors: Kimura Juki , Takahashi Satoru , Ogiso Tadashi , Yoshida Yasunori , Akagi Keisuke , Hasegawa Ryohei , Kurata Mitsuo , Hirose Masao , Shirai Tomoyuki
Published: August 2005

Modifying effects of dietary administration of the monoterpene d‐limonene were examined using a multi‐organ carcinogenesis model. Groups of twenty F344 male rats were treated sequentially with N‐diethylnitrosamine (DEN, i.p.), N‐methyl‐N‐nitrosourea (MNU, i.p.), 1,2‐dimethylhydrazine (DMH, s.c.), N‐butyl‐N‐(4‐hydroxybutyl)nitrosamine (BBN, in drinking water) and dihydroxy‐di‐N‐propylnitrosamine (DHPN, in drinking water) during the first 4 weeks (DMBDD treatment), and then (d‐limonene was administered in the diet, at the dose of 2.0, 1.0 or 0.5%. The maximal tolerable dose was 2.0% under the present conditions. Further groups were treated with DMBDD or 2.0%d‐limonene alone as controls. All surviving animals were killed at week 28, and major organs were examined histopathologically for development of preneoplastic and neoplastic lesions. The incidences and/or multiplicities of renal atypical tubules and adenomas were increased in animals fed 2.0%d‐limonene. The immunohistochemical reactivity for α2u‐globulin in the proximal tubules was greater in rats fed d‐limonene than in the carcinogen alone group. No enhancing or inhibitory effect was noted for tumor development in other organs. The present results indicate a lack of any chemopreventive effect of (d‐limonene in any organ of male rats under the present experimental conditions.
Changes in Cell Proliferative Parameters of SCCVII and EMT6 Murine Tumors after Single‐dose Irradiation
Authors: Murata Rumi , Nishimura Yasumasa , Shibamoto Yuta , Hiraoka Masahiro , Abe Mitsuyuki
Published: August 2005

To understand better the repopulation kinetics of tumor cells after radiotherapy, we Investigated changes in cell proliferative parameters after single‐dose irradiation of SCCVII tumors in C3H/He mice and EMT6 tumors in Balb/c mice. The following parameters were determined 0–15 days after single irradiation at 20 or 30 Gy; dividing fraction (DP), potential doubling time (Tpot), number of clonogenic cells per tumor (Ncln), and volume doubling time (Tvol). DF and Tpot were determined by in vivo‐in vitro cytokinesis‐block assay with cytochalasin B, Ncln was measured by in vivo‐in vitro colony‐forming assay, and Tvol was determined by growth delay assay. In both tumors, longer Tpot and lower DF and Ncln were obtained for 3–4 days after irradiation, but in SCCVII tumors these values returned to the pretreatment levels 9 days after irradiation. In EMT6 tumors, Tpot, DF, and Ncln did not return to the pretreatment levels even 12 days after irradiation. In the regrowth phase of both tumors following irradiation at 20 Gy, Tvol was longer than the pretreatment level, although Tpot was similar in SCCVII and only slightly longer in EMT6. Therefore, the cell loss factor in the regrowth phase was considered to be higher than the pretreatment level in both tumors. From the results, recruitment of previously quiescent cells into the proliferative pool in these tumors was suggested to contribute to repopulation after radiation.
Histologic Classification of Endoscopically Removed Flat Colorectal Polyps: A Multicentric Study
Authors: Rubio Carlos A. , Kato Yo , Hirota Teruyuki , Muto Tetsuichiro
Published: August 2005

A total of 594 flat colorectal polyps, removed at endoscopy, were histologically classified into non‐neoplastic (n=49) and neoplastic (n=545) polyps. Non‐neoplastic polyps were subdivided into metaplastic (n=45) and hyperplastic (n=4), whereas neoplastic polyps were subdivided into adenomas (n=481), intramucosal carcinomas (n=28) and invasive adenocarcinomas (n=36). Several adenoma phenotypes were discerned: tubular (n=375), serrated (n=59), villous (n=39), mixed (n=7) and fenestrated (n=1). Intramucosal carcinomas were subdivided into tubular (n=26) and serrated (n=2), and invasive adenocarcinomas into tubular (n=32), serrated (n=3) and fenestrated (n=1). The microscopic characteristics of each histologic phenotype described in this communication are defined and illustrated.
Anti‐tumor Effects of Hyperthermia Plus Granulocyte Colony‐stimulating Factor
Authors: Kokura Satoshi , Yoshikawa Toshikazu , Tainaka Kenzo , Kaneko Toshiro , Iinuma Syouji , Nishimura Shunichiro , Matuyama Kiichi , Kondo Motoharu
Published: August 2005

The role of neutrophils in the anti‐tumor effects of hyperthermia was investigated in an experimental rat model, and the efficacy of hyperthermia combined with recombinant human granulocyte colony‐stimulating factor (G‐CSF) was similarly investigated. AH109A carcinoma cells were transplanted into the hind legs of Donryu rats, then heated by a radio‐frequency dielectric heater. In this study, because the myeloperoxidase (MPO) activity of neutrophils was not affected by heating or G‐CSF, MPO activity was measured as an index of neutrophil migration into tumor tissue. After hyperthermia, MFO activity in tumor tissue increased significantly, suggesting migration of neutrophils into tumor tissue. Depletion of circulating neutrophils by the intraperitoneal injection of anti‐rat neutrophil antibody decreased the anti‐tumor effects of hyperthermia. Subsequently, we used hyperthermia plus intraarterial G‐CSF to enhance the anti‐tumor effect. Hyperthermia was induced 1 h after injection of G‐CSF, a time when MPO activity in tumor tissue was maximal. A satisfactory thermal effect was noted even in cases where tissue could not be heated sufficiently. In conclusion, neutrophils have an important role in the anti‐tumor effects of hyperthermia, and administration of G‐CSF enhances these effects.
Establishment and Characterization of Choroid Plexus Carcinoma Cell Lines: Connection between Choroid Plexus and Immune Systems
Authors: Enjoji Munechika , Iwaki Toru , Hara Hideo , Sakai Hironori , Nawata Hajime , Watanabe Takeshi
Published: August 2005

Murine choroid plexus cell lines were produced from choroid plexus carcinoma generated in transgenic mice harboring the viral oncogene simian virus 40 large tumor antigen under transcriptional control of an intronic enhancer region from the human immunoglobulin heavy chain (IgH) gene. Two morphologically distinct cell lines have been cloned. These established cell lines retained the characteristics of choroid plexus cells in that they expressed such choroid plexus cell marker or related proteins as transthyretin and α2‐macroglobulin. They were tumorigenic in nude mice. In the cell lines, the μA and μB (HE2) motifs within the IgH intronic enhancer were active and we also demonstrated the existence of the proteins binding to these motifs, suggesting a potential link between the choroid plexus and immune systems. It is considered that these binding proteins act as trans‐activators for the enhancer and may belong to the class of ETS‐related proteins. These cell lines and xenografts should be useful materials for analyses of choroid plexus functions.
A Comparative Study of Glioma Cell Lines for p16, p15, p53 and p21 Gene Alterations
Authors: Zhang Shujing , Endo Sumio , Koga Hisashi , Ichikawa Tomio , Feng Xuelian , Onda Kiyoshi , Washiyama Kazuo , Kumanishi Toshiro
Published: August 2005

A total of 10 glioma cell lines were examined for alterations of the p16, p15, p53 and p21 genes, which are tumor suppressor genes or candidates with direct or indirect CDK‐inhibitory functions. Genetic alterations (deletions or mutations) were frequently seen in the p16, p15 and p53 genes in these cell lines, but not in the p21 gene. When the states of the p16, p15 and p53 genes were compared among cell lines, all the cell lines showed abnormalities in at least 1 gene, often in 2 or 3 genes coincidentally, suggesting that dysfunction of these genes is closely related to glioma cell growth. Although alteration of all 3 genes was most frequent, there were cell lines having either p16/p15 or p53 or p16 and p53 gene alterations, suggesting that the time order of these genetic alterations was variable depending on the cell line. Among cell lines examined, one with homozygous p53 gene deletion seemed of particular practical value, since such a cell line might be useful in various studies, including investigation of the functions of various mutant p53 genes in the absence of heteromeric protein formation. On examination of the primary tumor tissues, the same alterations of the p16/p15 and p53 genes as detected in the cell lines were demonstrated in all 6 cases examined: p16/p15 gene deletion in 1, p16 gene mutation in 1 and p53 gene mutations in 5 cases. This suggested that the p16/p15 and the p53 gene alterations and their combinations in at least some glioma cell lines reflected those in the primary glioma tissues.
Glucocorticoid and Estrogen Receptors Have Elevated Activity in Human Endometrial and Ovarian Tumors as Compared to the Adjacent Normal Tissues and Recognize Sequence Elements of the H‐ras Proto‐oncogene
Authors: Zachos George , Varras Michael , Koffa Maria , Ergazaki Maria , Spandidos Demetrios A.
Published: August 2005

We examined the level of receptor binding in H‐ras elements, using nuclear extracts derived from human endometrial and ovarian lesions and from adjacent normal tissue in gel retardation assays. We found increased binding of the glucocorticoid receptor (GR) to the H‐ras GR element in more than 90% of endometrial tumors and in all ovarian tumors tested, as compared to the corresponding adjacent normal tissue. Additionally, we found elevated binding of the estrogen receptor (ER) in H‐ras ER element in all pairs of ovarian tumor/normal tissue tested, whereas in ER‐negative control breast tumor/normal tissue pairs, no differences in ER DNA‐binding levels were observed. These results suggest that steroid hormone receptor binding could directly activate the H‐ras oncogenic potency in human endometrial and ovarian lesions, providing additional evidence for the role of H‐ras expression in hormonally responsive human cancers.
Interleukin‐6 and Granulocyte Colony‐stimulating Factor Synergistically Increase Peripheral Blood Progenitor Cells in Myelosuppressive Mice
Authors: Suzuki Hideki , Okano Akira , Ejima Chieko , Konishi Atsushi , Akiyama Yukio , Ozawa Keiya , Asano Shigetaka
Published: August 2005

We previously reported a successful peripheral blood stem cell harvest by co‐administration of recombinant human (rh) interleukin‐6 (IL‐6) and rh granulocyte colony‐stimulating factor (G‐CSF) in normal mice. In the present study, to evaluate further the utility of this observation for autologous peripheral blood stem cell transplantation, we examined the effects of rhIL‐6 and rhG‐CSF on peripheral blood granulocyte‐macrophage colony‐forming units (CFU‐GM) in carboplatin (CBDCA)‐induced and irradiation‐induced myelosuppressive mouse models. After CBDCA administration, blood cell counts decreased to the nadir, and then recovered to a normal level. In this recovery phase, the peripheral CFU‐GM level increased to 3.8‐fold higher than the pretreatment level. Administration of rhIL‐6 (10 μg/day) alone induced a 40‐fold increase in peripheral CFU‐GM from the normal level at day 14. In combination with rhG‐CSF (0.35 μg/day), which alone induced a 74‐fold increase, rhIL‐6 synergistically increased the CFU‐GM level by 1200‐fold. In irradiated mice, similar results were observed. Administration of rhIL‐6 at 3 and 10 μg/day significantly increased CFU‐GM. Interestingly, in combination with rhG‐CSF, a lower dose of rhIL‐6 (1 μg/day) could induce CFU‐GM increase. We also examined CFU‐GM distribution in bone marrow, spleen and peripheral blood. Cytokine administration induced not only a change of CFU‐GM distribution, but also an increase in total CFU‐GM counts per mouse. These results suggest that co‐administration of rhIL‐6 and rhG‐CSF may be useful for autologous peripheral blood stem cell transplantation.
Inhibition of Growth and Metastasis of Ovarian Carcinoma by Administering a Drug Capable of Interfering with Vascular Endothelial Growth Factor Activity
Authors: Mu Jie , Abe Yoshiko , Tsutsui Tateki , Yamamoto Norihiko , Tai Xu‐Guang , Niwa Ohtsura , Tsujimura Takahiro , Sato Bunzo , Terano Hiroshi , Fujiwara Hiromi , Hamaoka Toshiyuki
Published: August 2005

The present study investigates the relationship between in vivo growth/metastasis of tumor cells and their capacity to produce the vascular endothelial growth factor (VEGF), as well as the regulation of tumor growth/metastasis using an angiogenesis‐inhibitory drug. Two cloned tumor cell lines designated OV‐LM and OV‐HM were isolated from a murine ovarian carcinoma OV2944. OV‐LM and OV‐HM cells grew in cultures at comparable rates. However, when transplanted s.c. into syngeneic mice, OV‐HM exhibited a faster growth rate and a much higher incidence of metastasis to lymph nodes and lung. Histologically, intense neovascularization was detected in sections of OV‐HM but not of OV‐LM tumor. OV‐HM and OV‐LM tumor cells obtained from in vitro cultures expressed high and low levels of VEGF mRNA, respectively. A difference in VEGF mRNA expression was much more clearly observed between RNAs prepared from fresh OV‐HM and OV‐LM tumor masses: RNA from OV‐HM contained larger amounts of VEGF mRNA, whereas RNA from OV‐LM exhibited only marginal levels of VEGF mRNA. An angiogenesis‐inhibitory drug, FR118487 inhibited the VEGF‐mediated in vitro growth of endothelial cells but did not affect the expression in vitro of VEGF mRNA by OV‐HM tumor cells. Intraperitoneal injections of FR118487 into mice bearing OV‐HM tumors resulted in: (i) a subsequent growth inhibition of primary tumors; (ii) a marked decrease in neovascularization inside tumor masses expressing comparable levels of VEGF mRNA to those detected in control OV‐HM masses; and (iii) almost complete inhibition of metastasis to lymph nodes and lung. These results indicate that growth/metastasis of tumor cells correlates with their VEGF‐producing capacity and that an angiogenesis inhibitor, FR118487, inhibits tumor growth and metastasis through mechanism(s) including the suppression of VEGF function in vivo.
Radioimmunotherapy for Pancreatic Carcinoma Using 131I‐Labeled Monoclonal Antibody Nd2 in Xenografted Nude Mice
Authors: Inui Akimasa , Chung Yong‐Suk , Sawada Tetsuji , Kondo Yasuyuki , Ho Jenny J. L. , Kim Young‐S. , Sowa Michio
Published: August 2005

We investigated the biodistribution, radiolocalization, and radioimmunotherapeutic potential of 131I‐labeled Nd2 in athymic nude mice bearing human pancreatic carcinoma xenografts. 131I‐Nd2 was accumulated at high levels in the tumor, in contrast to blood, liver, spleen, and other normal organs. The tumor was clearly delineated in scintigraphs. The volumes of tumors of mice injected with 7.4 MBq of 131I‐Nd2 were 80% less than those of tumors before injection of radiolabeled Nd2. Fibrous or vacuolar degeneration was seen in histological sections of tumors of 7‐week‐treated mice. The growth of tumors in mice treated with misonidazole, a hypoxic cell radiosensitizer, and then injected twice with 3.7 MBq of 131I‐Nd2 was suppressed over 7 weeks. Neither leucocytopenia nor thrombocytopenia was severe after injection of radiolabeled Nd2. Thus 131I‐labeled Nd2 may have clinical application in the radioimmunotherapy of pancreatic cancer.
Genetic Screening in Hereditary Multiple Endocrine Neoplasia Type 1: Absence of a Founder Effect among Japanese Families
Authors: Sakurai Akihiro , Katai Miyuki , Itakura Yasunori , Nakajima Koji , Baba Kiyoshi , Hashizume Kiyoshi
Published: August 2005

Ten Japanese families with hereditary multiple endocrine neoplasia type 1 (MEN1) were examined. Five DNA polymorphic markers on the long arm of chromosome 11 were analyzed for genetic screening of MEN1 in members of affected families, and disease carriers were identified before clinical manifestations. Unlike MEN1 families in Newfoundland or in Tasmania, no consistent haplotypes were segregated with the disease in the Japanese families when defined by 5 nearby markers. The identification of asymptomatic disease carriers is of substantial clinical importance for early management, genetic counseling and to avoid unnecessary screening for non‐disease carriers. However, genetic screening of family members by polymorphic markers could be useful only to each family, and no generally applicable markers were found for Japanese subjects with MEN1.
Subsite (cervix/endometrium)‐specific Risk and Protective Factors in Uterus Cancer
Authors: Hirose Kaoru , Tajima Kazuo , Hamajima Nobuyuki , Takezaki Toshiro , Inoue Manami , Kuroishi Tetsuo , Kuzuya Kazuo , Nakamura Shigeo , Tokudome Shinkan
Published: August 2005

In Japan the incidence of cervical cancer has been high, but has recently been decreasing gradually, while the incidence of endometrial cancer is running at lower levels but is gradually increasing. To clarify the common and/or specific risk and/or protective factors of cervical cancer (CC) in contrast with endometrial cancer (EC), a comparative case‐control study was conducted at the Aichi Cancer Center Hospital, Japan. In total, 556 CC cases and 145 EC cases were included and 26,751 women, confirmed as free of cancer, were chosen as the common control group. Odds ratio and its 95% confidence interval (95%CI) for each exposure variable were estimated by using an unconditional logistic regression model adjusted for age and first‐visit year. Habitual smoking and experience of pregnancy increased the risk of CC, while decreasing the risk of EC. Greater body mass index (>20), daily intake of fruit and more frequent intake of boiled or broiled fish (>1–2 times/week) decreased the risk of CC, whereas they increased the risk of EC. Daily intake of milk decreased the risk of CC. The results obtained from this study suggest that several EC‐increasing risk factors are in fact CC‐decreasing determinants. The observed risk reduction in both CC and EC by physical exercise and dietary control for health is noteworthy from the public health standpoint and warrants further investigation.
Inhibitory Effect of a Traditional Chinese Medicine, Juzen‐taiho‐to, on Progressive Growth of Weakly Malignant Clone Cells Derived from Murine Fibrosarcoma
Authors: Ohnishi Yasuharu , Fujii Hideki , Kimura Fuminari , Mishima Takaaki , Murata Jun , Tazawa Kenji , Fujimaki Masao , Okada Futoshi , Hosokawa Masuo , Saiki Ikuo
Published: August 2005

We have investigated the inhibitory effect of oral administration of Juzen‐taiho‐to, a Kampo (Chinese herbal) medicine, on progressive growth of a mouse fibrosarcoma. Spontaneously regressive QR‐32 tumor cells were able to grow progressively in vivo when coimplanted s.c. with a foreign body, gelatin sponge, whereas QR‐32 cells alone gradually grew for over 15 days after inoculation and thereafter regressed for up to 25 days. Oral administration of Juzen‐taiho‐to (40 mg/day/mouse) for 7 days after inoculation of QR‐32 cells with gelatin sponge resulted in significant inhibition of tumor growth and prolongation of the survival of the tumor‐bearing mice. This growth‐inhibitory effect of Juzen‐taiho‐to observed on day 25 was dose‐dependent over the dose range from 4 to 40 mg/day. Treatment with Juzen‐taiho‐to for 7 days before tumor inoculation with gelatin sponge also significantly suppressed tumor growth examined on day 25, as did the administration of bismuth subnitrate, which is well known to induce metallothionein, an antioxidant. On the other hand, inoculation of progressed tumor cells (QRsP) resulted in growth without gelatin sponge, leading to death in syngeneic mice. Administration of Juzen‐taiho‐to for 7 days after inoculation of QRsP cells resulted in a decrease of the tumor growth and prolongation of the survival of mice, but the effect was less than that on the growth of QR‐32 regressor tumor after coimplantation with gelatin sponge. These results suggest that the inhibitory effect of Juzen‐taiho‐to is partly associated with prevention of gelatin sponge‐elicited progressive growth, probably mediated by endogenous factors including antioxidant substances, in addition to the augmentation of host‐mediated antitumor activity.