bluuebunny/crossref_metadata_2025 huggingface dataset:
Relationships among Tenascin Expression, DNA Ploidy Patterns, and Multidrug Resistance Gene Product (P‐Glycoprotein) in Human Colon Carcinoma
Authors: Sugawara Isamu , Hirakoshi Jun , Kusakabe Moriaki , Masunaga Atsuko , Itoyama Shinji , Sakakura Teruyo
Published: August 2005

Relationships among tenascin expression, DNA ploidy, and P‐glycoprotein were examined in 81 primary human colon cancers and 61 metastatic lymph nodes. First, the DNA ploidy patterns of colon cancerous tissue surrounded (TN+) and not surrounded (TN−) by tenascin immunoreactivity were investigated. Then the expression of P‐glycoprotein, one of two multidrug resistance gene products, was examined in TN+ and TN− colon cancer tissues by immunohistochemistry. Aneuploid DNA patterns were observed at high frequency in TN colon cancer tissues (37/61) and metastatic lymph nodes (44/52). In contrast, diploid DNA patterns were observed predominantly in TN+ colon cancer tissues (50/56). Although P‐glycoprotein expression was observed in primary TN+ and TN− colon cancer (9/81), the level of P‐glycoprotein expression was not correlated with DNA aneuploidy in TN−colon cancer tissues. Overall, reduced tenascin expression was correlated well with DNA aneuploidy, but no significant correlation was found between DNA aneuploidy and P‐glycoprotein appearing when cancer cells become resistant to several anti‐cancer drugs. Thus, tenascin may play an important role in preventing colon cancer cells from invading surrounding tissues.
A Hospital‐based Case‐Control Study on Hepatocellular Carcinoma in Fukuoka and Saga Prefectures, Northern Kyushu, Japan
Authors: Fukuda Katsuhiro , Shibata Akira , Hirohata Itsuyo , Tanikawa Kyuichi , Yamaguchi Genjiro , Ishii Misa
Published: August 2005

Three hundred and sixty‐eight case‐control sets (male 287 pairs; female 81 sets) were collected for a hospital‐based case‐control study of primary hepatocellular carcinoma (HCC) conducted in Northern Kyushu, Japan. All incident cases of HCC were collected weekly from the inpatients (aged 40–69) of the Second Department of Internal Medicine, Kurume University Hospital between April, 1986 and May, 1992. One control for a male case and 4 controls for a female case were sampled, being matched to a case on age (same 5‐year age class), sex, residence (prefecture) and time of hospitalization (within 2 months after a case interview) from the inpatients of two general hospitals in Kurume. Information was collected by interview in person by a well‐trained interviewer and from a review of hospital records by the authors. Multivariatc analyses based on a conditional logistic regression model without an interaction term revealed that hepatitis B surface antigen (HBsAg) positive status (odds ratio (OR) = 8.67; 95% confidence Interval (95%CI) = 2.54–29.57), history of blood transfusion over 10 years previously (2.40; 1.26–4.56), parental history of hepatic diseases (2.31; 1.11–4.80) and heavy alcohol drinking (60≦ drink‐years) by age 40 (3.23; 1.61–6.51) were statistically significant risk factors of male HCC. Univariate analysis for females also showed an elevated OR of HBsAg (7.58; 1.96–29.35). Although the sample size was limited, univariate analysis indicated that anti‐hepatitis C virus antibody by c100‐3 antigen positive status had a statistically significant OR for HCC in both sexes.
HTLV‐I, HIV‐I, and Hepatitis B and C Viruses in Western Province, Papua New Guinea: A Serological Survey
Authors: Yamaguchi Kazunari , Inaoka Tsukasa , Ohtsuka Ryutaro , Akimichi Tomoya , Hongo Tetsuro , Kawabe Toshio , Nakazawa Minato , Futatsuka Makoto , Takatsuki Kiyoshi
Published: August 2005

Seven hundred and twenty‐three serum samples from individuals in 13 Gidra‐speaking villages in Western Province, Papua New Guinea were tested for evidence of infection with human T‐ lymphotropic virus type I (HTLV‐I), human immunodeficiency virus type I (HIV‐I), hepatitis B virus (HBV) and hepatitis C virus (HCV). No samples were positive for antibodies to HIV‐I. Antibodies to HTLV‐I were found in 13 samples (1.8%), HBV surface antigens (HBsAg) were found in 86 samples (11.9%), and antibodies to HCV were found in 30 samples (4.1%). Six (46.2%) of 13 HTLV‐I positive samples were positive for HCV or HBsAg. The seropositive rate varied in different villages and the incidence of HTLV‐I and HCV was higher in coastal and riverine areas than inland.
Loss of VLA‐2 Collagen Receptor in Breast Carcinoma, Facilitating Invasion and Metastasis
Authors: Arihiro Koji , Inai Kouki , Kurihara Kanji , Takeda Shinpei , Kaneko Mayumi , Kuroi Katsumasa , Toge Tetsuya
Published: August 2005

The integrin VLA‐2 as a collagen receptor and VLA‐5 as a fibronectin receptor were detected immunohistochemically in normal, benign tumor and carcinoma tissues of the breast. Both proteins were also detected by Western‐blot analysis in some carcinoma cases. Epithelial and myoepitheiial cells of both normal breast and benign tumor were in all cases immunoreactive for VLA‐2 in the plasma membrane. Carcinoma cells in the invasive component were not immunoreactive for VLA‐2 in 31 (46%) of 67 cases. Carcinoma cells in the intraductal components were negative for VLA‐2 in only 4 (11%) of 36 cases, while 20 cases (56%) showed weak expression and 12 cases (33%) showed strong expression. Metastatic carcinoma cells in the lymph nodes of 6 cases showed no immunoreactivity except in one case, whereas, again with the exception of one case, the carcinoma cells in the primary tumors did show VLA‐2 expression. With regard to VLA‐5, there was no difference in its expression in the invasive components and the intraductal components. These findings suggest that the loss of VLA‐2 plays a role in the invasion and metastasis of breast carcinoma.
Long‐term Preservation of Renin‐secreting Ability by Human Adult Juxtaglomerular Tumor Cells in Explant Culture
Authors: Armato Ubaldo , D'Agostino Domenico , Romano Flora , Salvetti Angelo , Mantero Franco
Published: August 2005

Studies on cultured human renin(R)‐producing tumors cells are few. In this work the R secretion by a human Juxtaglomerular tumor (JGT) in various tissue culture models was evaluated by a new immunoradiometric assay. Freshly isolated JGT cells actively secreted total R (tR; about 70% of which is proR) into the perfusion media of very short‐term cultures (tR concentration, 100–400 ng/ml/106 cells), independently of factors stimulating or inhibiting R output by normal JG cells. Primary monolayer cultures of the same JGT rapidly lost their tR‐secreting capability and died by apoptosis within two months. Conversely, a JGT explant survived for up to 22 months in vitro. During the first year of culture, this explant increased in volume and generated, at 3‐ to 4‐monthly intervals, several self‐limited cellular outgrowths, from which it became detached. Meanwhile, tR secretion by the explant decreased very slowly, though its decline was transiently and partly reversed by various combinations of growth factors, hormones, a prostaglandin, and selenous acid added to either a serum‐enriched or a synthetic medium. By the 12th month in vitro, tR secretion had faded away. Like the primary monolayers, the various explant outgrowths, once detached, stopped secreting tR and died in a few weeks. Hence, the preservation of a histiotypic relationship and the actions of several mitogenic and/or differentiating agents are essential for the long‐term survival and the continuance of R secretion by human JGT cells in vitro.
Head and Neck Carcinomas in Patients under the Age of Forty Years
Authors: Tsukuda Mamoru , Ooishi Kiminao , Mochimatsu Izumi , Sato Hirohisa
Published: August 2005

From 1973 to 1988, 1, 104 patients with head and neck carcinomas (excluding thyroid carcinoma) were admitted to the Yokohama City University Hospital, and among them, 48 (4.3%) were under the age of 40 years. We made a retrospective survey of these young patients. There were 36 males and 12 females. An increasing ratio of male to female was seen with increasing age. The primary site was most frequently the nasopharynx (29 cases), followed by the maxillary sinus (9 cases), and the tongue (4 cases). Squamous cell carcinoma in these young patients characteristically showed a low grade of differentiation.
A Novel Human Monoclonal Antibody Directed to a Tumor‐associated Antigen
Authors: Kudo Toshio , Kobayashi Syunsuke , Murakami Kazuhiro , Takano Ryo , Nose Masato , Sato Nobuyuki , Saeki Hisaaki , Saijyo Susumu , Hiyamuta Syuichi , Kadota Akihiko , Tachibana Takehiko
Published: August 2005

Twelve human monoclonal antibodies (HuMAb) were established by the fusion of (mouse × human) heteromyeloma cells with B‐lymphoblastoid cells derived from the regional lymph nodes of three patients with squamous cell carcinoma of the lung. They were tested for reactivity to two kinds of proteins (purified protein derivatives and bovine serum albumin) by ELISA, Sq‐19 (squamous cell carcinoma) culture cells by indirect membrane immunofluorescence tests, and Sq‐19 tumor xenograft by immunohistological study. Among them, one HuMAb 904F (IgM, λ) was selected. In indirect membrane immunofluorescence tests, this 904F antibody reacted with various kinds of cell lines, e.g. lung cancer, esophageal cancer, endometrial cancer, and stomach cancer. It did not react with malignant hematopoietic and diploid fibroblast cell lines. Immunohistologically, it stained the tumor nests of squamous cell carcinoma, adenocarcinoma, and large cell carcinoma of the lung. It also stained those of esophagus and colon, but not those of small cell carcinoma of lung, or stomach. On frozen‐section specimens of normal tissues from various organs, it showed only limited areas of positive staining. Limited positive findings were observed at a reticular zone of the adrenal gland, at the esophagus as weak staining, and at islets of the pancreas as very weak staining. Western blotting analysis demonstrated that it recognized a 54 kDa trypsin‐sensitive molecule which is expressed on the surface of tumor cells. These results suggest the 904F monoclonal antibody detects a novel tumor‐associated antigen which is recognized by the human immune system.
Generation of Monoclonal Antibodies to the Rabbit Interleukin‐2 Receptor a Chain (CD25) and Its Distribution in HTLV‐1‐transformed Rabbit T Cells
Authors: Kotani Masaharu , Yamamura Yasuko , Tsudo Mitsuru , Tamatani Takuya , Kitamura Fujiko , Miyasaka Masayuki
Published: August 2005

Rabbits can be infected with human retroviruses such as human T‐cell leukemia virus‐1 (HTLV‐1) and human immunodeficiency virus (HIV), and provide useful animal models to study retroviral diseases such as adult T‐cell leukemia and HIV. Previously we have succeeded in generating monoclonal antibodies (mAbs) against rabbit CD4, CD5 and CD11a antigens. To make this animal species more amenable to cellular and molecular studies, we have attempted to extend the panel of mAbs against rabbit CD antigens. Here we report on the generation of three neutralizing mAbs against interleukin‐2 receptor α chain (IL‐2Rα) (CD25), Kei‐α1 (IgG2b), Kei‐α2 (IgG2a) and Kei‐α3 (IgG1). They specifically recognize the rabbit Mr 55,000 IL‐2 binding protein, IL‐2Rα, and completely inhibit both high‐ and low‐affinity IL‐2 binding to F648b cells that express IL‐2Rα as well as IL‐2Rβ. The use of mAb Kei‐α1 confirmed that the rabbit IL‐2Rα is not only a low‐affinity IL‐2R on its own but also an essential component of high‐affinity IL‐2R as found in other animal species, and that rabbit activated T cells including HTLV‐1‐transformed cell lines express high levels of the IL‐2Rα. Together with mAbs against various rabbit CD antigens that we reported previously, these neutralizing mAbs to IL‐2Rα will be valuable for studies of human retrovirus infections, such as those induced by HTLV‐1 or HIV, in rabbits.
Effect of 5‐Fluorouracil and UFT on Experimental Liver Metastasis Model of Colorectal Cancer Using Mouse Colon 26 Cells
Authors: Tominaga Takeshi , Yoshida Yoko , Kosaki Goro , Kurihara Minoru
Published: August 2005

Effects of 5‐fluorouracil (5‐FU) and UFT on an experimental liver metastasis model were compared at equi‐effective dosage levels against subcutaneous tumor of mouse colon 26. 5‐FU at the dosage level of 40 mg/kg suppressed the subcutaneous tumor growth by 70.0% and 45.0% on day 13 and day 18, respectively, and UFT at 20 mg/kg provided almost equal suppression (63.0% and 48.0%). In the liver metastasis model, 5‐FU at 40 mg/kg showed more potent prevention of the formation of metastatic foci (94.9%) than did UFT (60.4%) at 20 mg/kg. 5‐FU at 40 mg/kg produced a much higher peak serum level of 5‐FU than did UFT at 20 mg/kg and also showed a much higher AUC (area under the curve) level in the portal blood. These results suggest that oral administration of 5‐FU might be useful in prevention of liver metastasis of colorectal cancer.
Enhancement of Antitumor Activity of Cisplatin on Human Gastric Cancer Cells in vitro and in vivo by Buthionine Sulfoximine
Authors: Saikawa Yoshiro , Kubota Tetsuro , Kuo Tsong‐Hong , Furukawa Toshiharu , Tanino Hirokazu , Watanabe Masahiko , Ishibiki Kyuya , Kitajima Masaki
Published: August 2005

An attempt was made to evaluate the enhancement of the antitumor activity of cisplatin (DDP) by buthionine sulfoximine (BSO) in vitro and in vivo. In the in vitro study, pre‐treatment with BSO (5, 10 and 25 mM) increased the antitumor activity of DDP against the gastric cancer cell lines MKN‐28 and MKN‐45, whereas BSO alone exhibited only slight antitumor activity (inhibition rate, 20–30%). In the in vivo study, the antitumor effects of DDP against human gastric cancer xenografts St‐15 and SC‐l‐NU in BALB/c nu/nu mice were enhanced pretreatment with BSO, which was administered intraperitoneally at a dose of 500 mg/kg according to a schedule of qd × 3. BSO alone showed no antitumor effects against these tumors in nude mice. The side effects (assessed in terms of death rate and body weight loss) associated with the maximum tolerated dose of DDP (9 mg/kg) were not increased by BSO pretreatment. As BSO increased the antitumor activity of DDP without a corresponding increment of its toxicity, BSO appears to be a promising agent for further study.
Synergistic Antitumor Effects of Carboplatin and Interferons on Hepatic Metastases of Colon Carcinoma 26 and M5076 Reticulum Cell Sarcoma
Authors: Iigo Masaaki , Shimizu Ishinori , Sagawa Katsuhiko
Published: August 2005

The effects of combination therapy including various antitumor agents and interferon on mice bearing hepatic metastases of colon carcinoma 26 were determined. Combined treatment with interferoh‐αA/D and carboplatin (CBDCA) was associated with a considerably more pronounced antitumor effect than was treatment with either drug alone. Marine interferon‐β and ‐γ each also potentiated the antitumor activity of CBDCA. Combination therapy with interferon‐αA/D and CBDCA also resulted in marked inhibition of hepatic metastasis of M5076 reticulum cell sarcoma. However, interferon‐β did not potentiate the antitumor activity of CBDCA against either subcutaneously implanted colon carcinoma 26 or pulmonary metastases of this tumor. Thus, in our model the combined administration of interferon and CBDCA was associated with a synergistic antitumor effect on hepatic metastases alone.
Inhibitory Effects of (—)‐Epigallocatechin Gallate on Spontaneous Hepatoma in C3H/HeNCrj Mice and Human Hepatoma‐derived PLC/PRF/5 Cells
Authors: Nishida Hiroshi , Omori Masahide , Fukutomi Yasushi , Ninomiya Mitsuo , Nishiwaki Shinji , Suganuma Masami , Moriwaki Hisataka , Muto Yasutoshi
Published: August 2005

The inhibitory effect of (—)‐epigallocatechin gallate (EGCG), a main constituent of Japanese green tea, on spontaneous hepatoma in C3H/HeNCrj mice was investigated. A total of 72 mice were divided into three groups; the control group without EGCG, and two experimental groups receiving 0.05% (w/w) or 0.1% EGCG in drinking water. EGCG reduced the incidence of hepatoma‐bearing mice from 83.3% (control) to 56.0% (0.05% EGCG) and 52.2% (0.1% EGCG), and also reduced the average number of hepatomas per mouse from 1.83 (control) to 0.72 (0.05% EGCG) and 0.91 (0.1% EGCG) at week 65. Ridit analysis of the distribution of the number of hepatomas in each group revealed that EGCG significantly increased the rate of mice without hepatoma in the two EGCG groups as compared to the control. EGCG did not affect body weight gain, food consumption or any serum biochemical parameter. EGCG inhibited the growth and secretion of α‐fetoprotein by human hepatoma‐derived PLC/PRF/5 cells without decreasing their viability. These results indicate that EGCG may be a practical, nontoxic preventive agent against human hepatoma.
Mortality among Inhabitants of an HTLV‐I Endemic Area in Japan
Authors: Iwata Kokichi , Ito Shin‐ichiro , Saito Hiroshi , Ito Mizuko , Nagatomo Masazumi , Yamasaki Tomihiro , Yoshida Shigeyuki , Suto Hisatsugu , Tajima Kazuo
Published: August 2005

A community‐based cohort study was conducted to clarify the risk of human T‐cell leukemia virus type I (HTLV‐I) infection for cause‐specific deaths. A total of 1,997 individuals (751 men and 1,246 women) aged 30 or older in A‐IsIand, Nagasaki Prefecture, Japan who had voluntarily attended annual mass health examinations, including serum HTLV‐I antibody test, were followed up for a mean period of 5.3 years. In a Cox proportional hazards analysis adjusted for age at baseline, the HTLV‐I seropositivity was found to be associated with mortality from all causes in men (hazard ratio (HR) 1.89; 95% confidence interval (CD 1.01–3.54) and women (HR 1.94; 95% CI 1.16–3.22). When the effects of 2 deaths (1 man and 1 woman) from adult T‐cell leukemia/lymphoma (ATL) were excluded, the mortality risk decreased slightly but was still significantly or marginally significantly greater than 1 in both men (HR 1.77; 95% CI 0.93–3.37) and women (HR 1.87; 95% CI 1.12–3.12). Further analysis of cause‐specific deaths revealed a significant increase in the risk for non‐neoplastic diseases but not for neoplasms excluding ATL. These findings suggest that long‐term HTLV‐I infection represents a health hazard greater than just that for the development of ATL. It was difficult, however, to draw a conclusion regarding the association between HTLV‐I infection and cancer risk, because the number of cancer deaths was small and the incidence of cancer was not investigated.
Growth and Differentiation Properties of Normal and Transformed Human Keratinocytes in Organotypic Culture
Authors: Tsunenaga Makoto , Kohno Yohko , Horii Izumi , Yasumoto Shigeru , Huh Nam‐ho , Tachikawa Tetsuhiko , Yoshiki Shusaku , Kuroki Toshio
Published: August 2005

The growth and differentiation of human normal keratinocytes and their transformed counterparts were examined in organotypic cultures in which the keratinocytes were grown at the air‐liquid interface on top of contracted collagen gel containing fibroblasts. We developed a modified culture procedure including the use of a mixed medium for keratinocytes and fibroblasts. Normal keratinocytes formed a three‐dimensional structure of epithelium that closely resembled the epidermis in vivo, consisting of basal, spinous, granular and cornified layers. Cells synthesizing DNA were located in the lowest basal layer facing the collagen gel. Expressions of proteins involved in epidermal differentiation were examined by immunohistochemical staining and compared with those in skin in vivo. In the organotypic culture, transglutaminase, involucrin and filaggrin were expressed, as in the epidermis in vitro, most prominently in the granular layer. Type IV collagen, a component of basement membrane, was expressed at the interface between the keratinocyte sheet and the contracted collagen gel. Keratinocytes transformed by simian virus 40 or human papilloma virus (HPV) exhibited a highly disorganized pattern of squamous differentiation. In particular, HPV‐transformed cells invaded the collagen gel. Organotypic culture is unique in that regulatory mechanisms of growth and differentiation of keratinocytes can be investigated under conditions mimicking those in vivo.
Frequent Loss of α Catenin Expression in Scirrhous Carcinomas with Scattered Cell Growth
Authors: Ochiai Atsushi , Akimoto Shingo , Shimoyama Yutaka , Nagafuchi Akira , Tsukita Shoichiro , Hirohashi Setsuo
Published: August 2005

To investigate the mechanisms of disruption of cell‐cell contact in scirrhous carcinoma cells, the expression of both E‐cadherin and α catenin, which is an intracellular cadherin‐binding molecule, were determined in scirrhous‐type adenocarcinomas of the stomach and breast using immunohistochemical and immunoblotting techniques. The losses of E‐cadherin expression in gastric and breast scirrhous adenocarcinomas were 18.1% and 0%, respectively, and those of α catenin expression were 54.6% and 75%, respectively. Frequent loss of α catenin expression occurred in scirrhous carcinomas with scattered cell growth in the stomach and the breast and showed no organ specificity. In addition, all the infiltrating lobular carcinomas, which also infiltrate the stroma as single cells, showed no E‐cadherin or α catenin expression. These findings suggest that down‐regulation of either α catenin or E‐cadherin plays a critical role in the disruption of cell adhesion in carcinomas with scattered cell growth.
Inhibition of Mammary Tumors by Pretreatment with 17β‐Estradiol in F344 Rats Induced with N‐Methyl‐N‐nitrosourea
Authors: Ito Akihiro , Okamoto Taro , Fujimoto Nariaki , Ogundigie Peter Osa , Watanabe Hiromitsu
Published: August 2005

The influence of 17β‐estradiol (E2) and prolactin was studied on N‐methyl‐N‐nitrosourea (MNU)‐induced mammary carcinomas (MCAs) in rats. MNU was intravenously injected once into seven‐week‐old female F344 rats at a dose of 50 mg/kg body weight. Groups of rats also received either 2.5 mg of E2 or a continuous supply of prolactin and/or growth hormone via transplanted MtT/F84 (mammo‐somatotropic pituitary tumor). Rats were observed for up to 36 weeks after MNU administration. Although simultaneous administration of MNU and E2 did not much affect the occurrence of MCAs as compared to administration of MNU alone, rats treated with 2.5 mg of E2 for two weeks before MNU administration had significantly reduced occurrence of MCAs compared to those given MNU alone. In contrast, rats with MNU plus MtT/F84 showed high incidence and shortened latency of MCAs and they also had a high incidence of clitorial gland hyperplasias. Average pituitary weights and serum prolactin levels in E2‐treated rats were greatly increased compared to those of MNU‐alone rats. Average serum E2 levels were about 100 ng/ml in E2‐treated rats and 0.05 ng/ml in rats without E2 treatment. Serum prolactin levels were greatly increased in rats with MtT/F84. The results indicated that pretreatment with E2 before MNU administration was inhibitory while increased prolactin caused by grafting MtT/F84 after MNU injection was promotive for the occurrence of MCAs in female F344 rats.
Characterization of a Taxol‐resistant Human Small‐cell Lung Cancer Cell Line
Authors: Ohta Sei , Nishio Kazuto , Kubota Naohiro , Ohmori Tohru , Funayama Yasunori , Ohira Tatsuo , Nakajima Hiroaki , Adachi Mitsuru , Saijo Nagahiro
Published: August 2005

Taxol is a novel anticancer agent with activity against a broad range of tumors. It has a unique ability to stabilize polymerized tubulin into microtubule bundles within the cell. We have established a taxol‐resistant human small‐cell lung cancer cell line (H69/Txl) by exposing H69 cells to stepwise increases in taxol concentration. The resistance of H69/Txl cells to taxol was 4.7‐fold that of the original H69 cells: the IC50 values for H69 and H69/Txl were 113.7 ± 56.54 nM and 538.7 ± 214.7 nM by the tetrazolium dye assay, respectively. Removal of the drug from the medium resulted in a 38% decrease in the growth rate of H69/Txl as compared with that in the presence of 30 nM taxol, suggesting that the growth of H69/Txl was partially dependent on taxol. H69/Txl showed higher sensitivity to vinca alkaloids such as vindesine, vincristine and vinblastine than the parental H69. There was no significant difference in intracellular [3H]taxol content between H69 and H69/Txl cells. No MDR‐1 mRNA was detected in H69/Txl by the reverse transcription polymerase chain reaction. There was no significant difference of total and polymerized tubulin content between H69 and H69/Txl cells. Altered mobility of one of the α‐tubulin isoforms in H69/Txl was revealed by using isoelectric focusing and Western blotting with anti‐α‐tubulin antibody. In H69, two α‐tubulin isoforms were observed, whereas three were evident in H69/Txl, two of them comigrating with the isoforms of H69 and the other being more acidic. We observed the increased acetylation of α‐tubulin in H69/Txl cells as compared with that in H69 cells. The acetylation of α‐tubulin may be responsible for the taxol resistance and/or taxol‐dependent growth of H69/Txl.
Opposing Effects of Indomethacin and Nordihydroguaiaretic Acid on Macrophage Function and Tumor Growth
Authors: Suzuki Michihiro , Li Jey , Asakura Toshio , Arai Kazuhiko
Published: August 2005

The effects of indomethacin (INDO) and nordihydroguaiaretic acid (NDGA) were compared on macrophage function and tumor growth. Intraperitoneal injections (5 mg/kg/day, 6, 8 or 9 days) of INDO in C57BL/6 mice stimulated the cytotoxicity of peritoneal macrophages and inhibited growth of subcutaneous B16 melanoma, whereas NDGA injections suppressed macrophage cytotoxicity, increased macrophage prostaglandin‐E2 (PGE2) release, and enhanced the tumor growth. Further, pretreatment of macrophages in vitro with the INDO group serum increased cytostasis against B16 tumor cells, while the use of NDGA group serum reduced it. Incubation of tumor‐bearer's macrophages in vitro with INDO (10‐6M and 10‐7M) or 10‐6M NDGA for 4 h reduced PGE2 release, but 10 ‐7M NDGA markedly enhanced the PGE2 release. Our data indicate that INDO and NDGA are able to modulate the macrophage function directly, and produce opposing effects on macrophage function and tumor growth. Inhibitory actions of INDO and NDGA on the cyclooxygenase or lipoxygenase pathway of arachidonic acid metabolism appear to contribute to their effects.
Enhancement of Blood Stasis and Vascular Permeability in Meth‐A Tumors by Administration of Hyperthermia in Combination with Tumor Necrosis Factor
Authors: Umeno Hiroshi , Watanabe Naoki , Yamauchi Naofumi , Tsuji Naoki , Okamoto Tetsuro , Niitsu Yoshiro
Published: August 2005

Blood stasis and vascular permeability induced by tumor necrosis factor (TNF) in Meth‐A tumors transplanted in BALB/c mice were significantly enhanced by hyperthermia at 40°C for 30 min immediately following TNF administration. A dose‐dependent, sustained decline in the intratumoral blood flow rate occurred following the administration of TNF alone (i.v.; 5 × 103, 5 × 104, or 5 × 105 JRU/kg) and was enhanced by the administration of hyperthermia in combination with the TNF, even though no decline occurred with hyperthermia alone. The combination of TNF at 5 × 105 JRU/kg and hyperthermia resulted in a blood flow ratio (ratio of blood flow after administration to that before) of 0.47 at 1 h compared with a ratio of 0.65 at 1 h after TNF alone. The blood flow in normal skin sites did not decrease in any case. The permeability of the intratumoral vasculature similarly increased in a dose‐dependent manner after the administration of TNF alone and was further increased by combination with hyperthermia, even though no increase occurred with hyperthermia alone. The mean permeability in mice receiving TNF alone at 5 × 105 JRU/kg was 1.35 times that in untreated mice. In mice receiving TNF at the same dose together with hyperthermia, the ratio was increased to 1.65. The results suggest that TNF selectively suppresses intratumoral blood flow, that this effect is enhanced by mild hyperthermia, and that the mechanism of the suppression by TNF with or without hyperthermia partly involves an increase in blood vessel permeability.
Identification of 29 Rat Genetic Markers by Arbitrarily Primed Polymerase Chain Reaction
Authors: Canzian Federico , Ushijima Toshikazu , Toyota Minoru , Hosoya Yoko , Sugimura Takashi , Nagao Minako
Published: August 2005

The number of genetic markers for the rat is still limited, in spite of its wide use in cancer research. To facilitate accurate mapping of both established and novel rat genetic markers, we constructed a linkage map by genotyping 105 F2 rats from ACI/N (ACI) and BUF/Nac (BUF) crosses. This map consists of 120 genetic markers that had been previously reported, mainly by two research groups, but had not been integrated. To find new genetic markers, the arbitrarily primed polymerase chain reaction (AP‐PCR) was applied to detect polymorphic bands between ACI and BUF rats. After testing 56 single primers and 12 combinations of primers, we found 36 bands produced by 16 single primers and two combinations to be reliably polymorphic between ACI and BUF rats. The 36 bands were typed in the 105 F2 rats, and 29 of them could be linkage‐mapped. AP‐PCR is thus useful to detect new genetic markers in laboratory strains of rats.
A Pilot Study for a Randomized Controlled Trial to Prevent Gastric Cancer in High‐risk Japanese Population: Study Design and Feasibility Evaluation
Authors: Tsugane Shoichiro , Tsubono Yoshitaka , Okubo Shunji , Hayashi Masato , Kakizoe Tadao
Published: August 2005

Observational epidemiological studies suggest that some nutrients reduce the risk of gastric cancer and that individuals with atrophic gastritis are at high risk of developing gastric cancer. One possible measure for gastric cancer prevention is therefore nutritional supplementation for the high risk group. Before recommending this strategy for the general public, however, a randomized controlled trial (RCT) is necessary. To evaluate the feasibility of an RCT, the authors conducted a pilot study using recipients of a health check‐up program in a general hospital in Japan. The subjects who were asked to participate in the trial had been diagnosed as having atrophic gastritis on the basis of serum pepsinogen I <70 ng/ml and the ratio of pepsinogen I to II <3.0. They were requested to ingest double‐blinded capsules containing different levels of vitamin C and β‐carotene every day. Out of the 219 subjects (118 males, 101 females) who were eligible for the study and had the required pepsinogen measurement, 90 (41%) met the criteria for atrophic gastritis. Among them, 55 (61%) (35 males, 20 females) gave their informed consent to participate in the RCT. Fifty‐four participants completed a 3‐month course of supplementation, and all of them agreed to a 5‐year supplementation period. The authors concluded that an RCT using double‐blinded nutritional supplements and targeting apparently healthy individuals is feasible in an intervention study for cancer prevention in Japan.
Prevention of Mammary Tumorigenesis in Acatalasemic Mice by Vitamin E Supplementation
Authors: Ishii Kunihiko , Zhen Li‐Xue , Wang Da‐hong , Funamori Yuka , Ogawa Katsuo , Taketa Kazuhisa
Published: August 2005

Adult male and female acatalasemic (C3H/AnLCsbCsb), hypocatalasemic (C3H/AnLCscCsc) and normal mice of C3H strain fed on regular laboratory chow for 15 months showed an increased incidence of spontaneous mammary tumor in the decreasing order of female acatalasemic, male acatalasemic, female hypocatalasemic and male hypocatalasemic mice. Normal mice did not develop mammary tumor. We conducted a prospective study with female acatalasemic mice, which showed the highest incidence of mammary tumor, to examine the preventive effect of vitamin E on mammary tumor. Female acatalasemic mice were fed on vitamin E‐deficient (28 animals) and vitamin E‐supplemented diet (25 animals) for 29 months. The incidence of mammary tumor in mice given the vitamin E‐supplemented diet was 47%, while that in mice given vitamin E‐deficient diet was 82% (P<0.002). Mammary tumors were apparent after 9 months of vitamin E deprivation and after 14 months of vitamin E supplementation. Female normal mice did not develop mammary tumor during a comparable period of time. The mean catalase activity of mammary gland in acatalasemic mice was 18.8% of that in normal mice. The results indicate that vitamin E protects acatalasemic mice against the development of mammary tumor.
Primary Malignant Lymphoma of the Brain: Demonstration of Frequent p16 and p15 Gene Deletions
Authors: Kumanishi Toshiro , Zhang Shujing , Ichikawa Tomio , Endo Sumio , Washiyama Kazuo
Published: August 2005

The p16 (MTS‐1) gene, a candidate tumor suppressor gene, was examined by means of Southern blot, PCR‐SSCP (polymerase chain reaction‐single‐strand conformation polymorphism) and nucleotide analyses in 5 cases of primary malignant lymphoma of the brain. By Southern blot analysis, the p16 gene was found to be deleted in at least 4 cases, homozygously (3 cases) or hemizygously (1 case). The p15 (MTS‐2) gene, another candidate tumor suppressor gene located in the vicinity of the p16 gene, to which it shows structural and functional similarity, was also found to be deleted in 4 cases. Our frequent detection (80%) of p16 and p15 gene deletions might suggest that these deletions are closely related to carcinogenesis in primary malignant lymphoma of the brain. SSCP and nucleotide analyses revealed no mutations of the p16 gene in any of the cases.
Immunocytochemical Detection of p53 in Cultures of Exfoliated Cells from Urine of Patients with Urothelial Cancers
Authors: Okuno Hiroshi , Kakehi Yoshiyuki , Ogawa Osamu , Yamada Hitoshi , Özdemir Enver , Okada Yusaku , Yoshida Osamu
Published: August 2005

Early diagnosis of urothelial cancer is critical for successful treatment. Mutation of the p53 gene together with allelic loss of chromosome 17p correlates well with high grade and invasiveness of urothelial cancer. Moreover, this mutation is reported to be an early event for carcinoma in situ of the urothelium. In order to develop a new non‐invasive diagnostic method for urothelial cancer, we have established a short‐term culture system for urinary exfoliated cells from patients with urothelial cancer. Immunocytochemical detection of p53 in these urine‐derived cells was conducted. Short‐term cultures of exfoliated cells from 50 ml samples of urine from 52 patients with urothelial cancers were made. Adequate cell growth (>105 cells per flask) was followed by passage onto glass chamber slides for p53 immunocytochemical staining. Successful passage was obtained in 40 of the 52 (76.9%) patients with urothelial cancers studied. The success rate for patients with tumors immunohistochemically positive for p53 nuclear accumulation was 90.5%, and 61.3% for those with tumors negative for p53 (P<0.05). Results of immunochemical analyses of the p53 in the urine cells and those in the tumor samples were identical in 92.1% of the patients. Culture of exfoliated cells from urine would be a good, non‐invasive method for the molecular diagnosis of urothelial cancer that should prove useful for the early detection and follow‐up of tumors with p53 mutation.
Triamterene Suppresses Bombesin‐enhanced Peritoneal Metastasis of Intestinal Adenocarcinomas Induced by Azoxymethane
Authors: Iishi Hiroyasu , Tatsuta Masaharu , Baba Miyako , Uehara Hiroyuki , Nakaizumi Akihiko , Akedo Hitoshi
Published: August 2005

The effects of combined administration of bombesin and the diuretic triamterene on the incidence of peritoneal metastasis of intestinal cancers induced by azoxymethane (AOM) and the labeling index of intestinal cancers were investigated in male inbred Wistar rats. From the start of the experiment, rats were given weekly s.c. injections of AOM (7.4 mg/kg body weight) for 10 weeks and s.c. injections of bombesin (40 μg/kg body weight) every other day, and from week 16, s.c. injections of triamterene (10 and 20 mg/kg body weight) every other day until the end of the experiment in week 45. Bombesin significantly increased the incidence of intestinal tumors and cancer metastasis to the peritoneum in week 45. It also significantly increased the labeling index of intestinal cancers. Although administration of both doses of triamterene with bombesin had little or no influence on the enhancement of intestinal carcinogenesis by bombesin, or the location, histologic type, depth of invasion, or labeling index of intestinal cancers, it significantly reduced the incidence of cancer metastasis. These findings indicate that triamterene suppresses cancer metastasis through a mechanism that does not affect the proliferation of intestinal cancers.
Antiproliferative Effect of a Novel Cholecystokinin‐B/Gastrin Receptor Antagonist, YM022
Authors: Murayama Tohru , Matsumori Yoshinobu , Iwata Nobuko , Ito Mitsuhiro , Taniguchi Taizo , Chihara Kazuo , Matsui Toshimitsu
Published: August 2005

Cholecystokinin (CCK)‐B and gastrin receptors are expressed on a variety of human tumor cells. Recently, we have demonstrated that the human brain CCK‐B receptors are identical to the gastrin receptors derived from the stomach mucosa, and that the brain‐gut peptides, CCK‐8 and gastrin I are mitogenic for mouse NIH 3T3 fibroblasts expressing human CCK‐B/gastrin receptors (N‐hCCKBR). In this report, we evaluated the antiproliferative potency of CCK‐B/gastrin receptor antagonists by using N‐hCCKBR cells. Among several antagonists, a benzodiazepine derivative, YM022 had the most potent activities in competing with [125I]CCK‐8 or [125I]gastrin I binding, inhibition of CCK‐8‐ or gastrin I‐induced phosphoinositide hydrolysis and increasing cytoplasmic free calcium. Interestingly, a potent antagonist for rat CCK‐B/gastrin receptors did not have such activities in N‐hCCKBR cells. YM022 inhibited the CCK‐8‐ or gastrin I‐induced [methyl‐3H]thymidine incorporation of N‐hCCKBR cells in a dose‐dependent manner. In the absence of exogenous peptide ligands, YM022 also inhibited the proliferation of several human cancer cell lines expressing the genes for both gastrin and its receptor. These results suggest that YM022 could intervene in the autocrine stimulation of human tumor cell lines through CCK‐B/gastrin receptors. N‐hCCKBR cells are an excellent tool to screen for novel human CCK‐B/gastrin receptor antagonists possessing antiproliferative activity for human cancer cells.
Induction of MAGE Genes in Lymphoid Cells by the Demethylating Agent 5‐Aza‐2′‐deoxycytidine
Authors: Shichijo Shigeki , Yamada Akira , Sagawa Kimitaka , Iwamoto Osamu , Sakata Motoko , Nagai Kojiro , Itoh Kyogo
Published: August 2005

MAGE genes encoding tumor antigens recognized by cytotoxic T lymphocytes are appropriate target molecules for specific immunotherapy of cancer. We have investigated whether the demethylating agent 5‐aza‐2′‐deoxycytidine (DAC) induces MAGE‐1, ‐2, ‐3, and ‐6 in normal and malignant lymphoid cells. DAC induced these MAGE genes in both PHA/interleukin‐2 (IL‐2)‐activated T cells from healthy donors and MAGE‐negative T and B cell leukemias in most cases. It also induced MAGE‐1 in IL‐2‐dependent T cell clones and all MAGE genes tested in Epstein‐Barr virus‐transformed B cell lines. Expression of MAGE‐1 protein in the cells was confirmed by western blot analysis with anti‐MAGE‐1 polyclonal antibody. Therefore, demethylation is a potent stimulus to induce MAGE genes in both normal and malignant lymphoid cells.
Overexpression of Multidrug Resistance Protein Gene in Human Cancer Cell Lines Selected for Drug Resistance to Epipodophyllotoxins
Authors: Koike Koji , Abe Tatsuya , Hisano Terumasa , Kubo Takeshi , Wada Morimasa , Kohno Kimitoshi , Kuwano Michihiko
Published: August 2005

Overexpression of either the multidrug resistance 1 (MDR1) gene or multidrug resistance protein (MRP) gene is involved in acquisition of multidrug‐resistant phenotypes in human cancer cells. In this study we examined whether selection for resistance to the epipodophyllotoxins, etoposide/teniposide (VP16/VM26), could induce overexpression of MDR1 or MRP. We have previously isolated two VP16/VM26‐resistant KB cell lines. Two VP16/VM26‐resistant KB cell lines, KB/VM‐1 and KB/VM‐4, which were selected by stepwise exposure to VM26 had decreased accumulation of [3H]VP16 and increased levels of MRP, but no apparent expression of MDR1 gene was observed. Another VP16/VM26‐resistant KB cell line, KB/VP‐4, which was further isolated from a VP16‐resistant KB cell line, KB/VP‐2, had decreased accumulation of [3H]VP16 and showed overexpression of MRP gene, but not that of MDR1 gene. We also isolated a VP16‐resistant cell line, IN157/VP‐1, from a human glioma cell line IN157. IN157/VP‐1 cells showed decreased accumulation of [3H]VP16 and overexpression of MRP gene, but not of MDR1. These findings suggest that selection for resistance to VP16/VM26, preferentially induces overexpression of MRP gene.
Reproducibility of Diagnosis and Its Influence on the Distribution of Lung Cancer by Histologic Type in Osaka, Japan
Authors: Yamamoto Seiichiro , Sobue Tomotaka , Yamaguchi Naohito , Yamamoto Satoru , Kikui Masanori , Kusunoki Yoko , Oshima Akira , Hanai Aya
Published: August 2005

The histologic types of lung cancer cases diagnosed in 1979–1980 (n=799) and 1987 (n=587) were independently reviewed by two pathologists in order to investigate the reproducibility of the diagnosis of the histologic type when the WHO classification (1981) was used. The specimens from 354 surgical cases and biopsy or cytology specimens from 1032 non‐surgical cases were reviewed. The inter‐observer agreement was 87.9% (k=0.79) for surgical cases and 81.4% (k=0.72) for non‐surgical cases. When compared to the original diagnosis, the agreement was 86.8% (k=0.78) for surgical and 86.4% (k=0.79) for non‐surgical cases in 1979–1980 and the agreement was 92.8% (k=0.87) for surgical and 89.1% (k=0.83) for non‐surgical cases in 1987. By histologic type, no difference in the agreement was observed except for large cell carcinoma. The distribution of histologic types after the review differed only slightly (less than 6%) from the original distribution. This suggests that in Osaka, Japan, the diagnosis based on the WHO classification (1981) had only a limited influence on the distribution of histologic types, and is not a major reason for the changing trends in lung cancer incidence by histologic type.
Role of Human Telomerase Reverse Transcriptase and Telomeric‐repeat Binding Factor Proteins 1 and 2 in Human Hematopoietic Cells
Authors: Yamada Koji , Yajima Tomomi , Yagihashi Atsuhito , Kobayashi Daisuke , Koyanagi Yoko , Asanuma Koichi , Yamada Mikako , Moriai Ryosuke , Kameshima Hidekazu , Watanabe Naoki
Published: August 2005

Telomerase, an enzyme that adds hexameric repeats of 5′‐TTAGGG‐3′, termed telomeres, to the ends of chromosomal DNA, has been implicated in cellular immortalization and cellular senescence. Recently several relevant genes have been cloned, including those encoding three major components of human telomerase: human telomerase RNA component (hTR), human telomerase reverse transcriptase (hTERT), and telomerase‐associated protein‐1 (TEP1). Also important are genes encoding human telomeric‐repeat binding factor proteins (TRF) 1 and 2. We compared 10 human malignant hematopoietic cell lines, 19 samples from patients with acute leukemia and normal granulocytes and monocytes to study telomerase activity and expression of these various genes using a reverse transcription‐polymerase chain reaction (RT‐PCR). In all 10 malignant cell lines with telomerase activity, hTR, hTERT mRNA, and TEP1 mRNA were expressed, while in normal monocytes and granulocytes without telomerase activity, expression of hTR, but not hTERT mRNA was detected. TEP1 mRNA was expressed in normal monocytes, but not granulocytes. Expression of TRF1 and TRF2 mRNAs was greater in the normal cells than in human malignant hematopoietic cell lines and in 16 samples of patients with acute leukemia. When differentiation of the malignant hematopoietic cell line HL‐60 was induced using tumor‐necrosis‐factor 471 and all‐trans retinoic acid (ATRA), telomerase activity decreased gradually during differentiation. Of the three telomerase components, only hTERT mRNA expression showed changes paralleling telomerase activity, becoming undetectable with differentiation. In contrast, initially low expression of TRF1 and TRF2 mRNAs increased during differentiation. Not only hTERT, but also TRF1 and TRF2 are important regulators of telomerase activity that represent potential targets for gene therapy against cancer.
Renal Carcinogenicity of Concurrently Administered Fish Meal and Sodium Nitrite in F344 Rats
Authors: Furukawa Fumio , Nishikawa Akiyoshi , Ishiwata Hitoshi , Takahashi Michihito , Hayashi Yuzo , Hirose Masao
Published: August 2005

The effects of long‐term concurrent administration of powdered fish meal and sodium nitrite were examined in F344 rats. A total of 600, 6‐week‐old rats were divided into 6 male and 6 female groups, each consisting of 50 animals. Rats in groups 1–3 and 7–9 were respectively fed diets supplemented with 64%, 32% and 8% (basal diet) fish meal, and simultaneously given 0.12% sodium nitrite in their drinking water. Groups 4–6 and 10–12 were respectively given 64%, 32% and 8% fish meal and tap water. At the 104th week, all surviving animals were killed and examined histopathologically. Treatment with fish meal dose‐dependently increased the incidences and multiplicities of atypical tubules, adenomas and renal cell carcinomas in sodium nitrite‐treated males. Females were less susceptible than males for renal tumor induction. In males given the 64% fish meal diet alone, the incidence and multiplicity of atypical tubules were also significantly increased as compared with the 8% fish meal alone case. Nephropathy was apparent in fish meal‐treated groups in a clear dose‐dependent manner, irrespective of the sodium nitrite treatment, and was more prominent in males than in females. Dimethylnitrosamine was found in the stomach contents after 4‐week treatment with 64% fish meal plus 0.12% sodium nitrite, at a level twice that in the 8% fish meal plus 0.12% sodium nitrite group. The results clearly indicate that concurrent administration of fish meal and sodium nitrite induces renal epithelial tumors. Further studies are required to elucidate how nephropathy and nitrosamines produced in stomach contents may contribute to the observed renal tumor induction.
L‐myc Restriction Fragment Length Polymorphism in Japanese Patients with Esophageal Cancer
Authors: Shibuta Kenji , Inoue Hiroshi , Sato Koichi , Matsuyama Ayumi , Ueo Hiroaki , Tanaka Youichi , Mafune Kenichi , Barnard Graham F. , Mori Masaki
Published: August 2005

L‐myc polymorphism is a representative genetic trait related to an individual's susceptibility to several cancers. However, there have been no reports concerning the association between esophageal cancer and L‐myc polymorphism. To analyze the distribution of polymorphism in Japanese patients with esophageal cancer, a molecular genotyping method using a polymerase chain reaction‐based restriction fragment length polymorphism (PCR‐RFLP) was used. Based on an analysis of 65 Japanese patients with esophageal cancer and 107 healthy control subjects, a significant difference was observed in either the distribution of genotypes (P=0.012) or of allele frequencies between the two groups (P 0.004). The relative risk of esophageal cancer for genotypes including the shorter allele was 2.9 compared to the longer allele homozygote. Furthermore, the patients with S‐allele had a tendency for poor prognosis among those with three genotypes. A significant difference between the distribution of genotypes and the incidence of lymph node metastasis was found based on the clinicopathological features of the cancers. These results suggest that L‐myc polymorphism may be implicated as a genetic trait affecting an individual's susceptibility to esophageal cancer, at least among Japanese patients.