bluuebunny/crossref_metadata_2025 huggingface dataset:
Polypoid Carcinoma of the Esophagus
Authors: Mori Masaki , Mimori Koshi , Sadanaga Noriaki , Watanabe Masayuki , Kuwano Hiroyuki , Sugimachi Keizo
Published: August 2005

Polypoid carcinoma of the esophagus is rare and little is known about its clinical and histopathologic features. We reviewed 500 surgical cases of esophageal carcinoma and analyzed 12 polypoid carcinomas. Clinical records were reviewed. Histologic examination was done on an average of 68 sections in each tumor. Immunohistochemical examination for proliferating cell nuclear antigen (PCNA) was done in selected sections. No special findings were seen with respect to age, sex, symptoms, or tumor location. The tumors, however, had several interesting features: 1) the main histologic type was squamous cell carcinoma, but other histologic features such as so–called carcinosarcoma, adenoid cystic carcinoma, and verrucous carcinoma were occasionally seen, 2) bidirectional differentiation to squamous and adcnocarcinomatous components was recognized, 3) intraepithelial spreading of the carcinoma was often present, 4) depth of invasion in the wall was often shallow, and 5) the prognosis was relatively good. The PCNA labeling index was well correlated with lymphatic or blood vessel permeation.
Comparative DNA Analysis by Image Cytometry and Flow Cytometry in Non‐small Cell Lung Cancer
Authors: Yamamoto Tatsuo , Horiguchi Hisashi , Kamma Hiroshi , Noro Masahiro , Ogata Takesaburo , Inage Yoshihisa , Akaogi Eiichi , Mitsui Kiyofumi , Hori Motokazu , Isobe Masaya
Published: August 2005

To determine whether image cytometry (ICM) is advantageous for clinical DNA analyses of tumor cells, nuclear DNA contents measured by ICM were compared with those by flow cytometry (FCM), using 46 samples of non‐small cell lung cancers. ICM was performed on smear specimens of fresh materials (f‐ICM) and cell suspensions obtained from paraffin‐embedded tumors (p‐ICM). The same cell suspensions were also analyzed by FCM (p‐FCM). Aneuploid rates/coefficient of variation (CV) of f‐ICM, p‐ICM, and p‐FCM were 76.1/4.90, 71.7/5.01 and 60.9/5.31%, respectively. There was a high correlation in the DNA indices between p‐ICM and p‐FCM (r=0.80). In the comparative DNA analysis, there were seven discordant samples. Six of them were estimated as aneuploid by p‐ICM, but they were miscounted as diploid or undefinablc (impossible) by p‐FCM. This was caused by measuring condensed nuclei or debris. All “impossible” samples in p‐FCM were squamous cell carcinoma with necrosis. In cell cycle analysis, the S and S+G2/M phase fractions in diploid samples were higher in p‐ICM than those in p‐FCM (P< 0.005), because the GO/G1 phase (2N) fraction presented by FCM was composed of cancer and non‐malignant cells in diploid cancers. In ICM, they can be separately measured by means of morphological selection. These findings indicated that ICM is superior to FCM, especially for the practical DNA measurement of a few cancer cells and in the evaluation of the proliferation rates.
Clinical Usefulness of CYFRA Assay in Diagnosing Lung Cancer: Measurement of Serum Cytokeratin Fragment
Authors: Sugama Yasuo , Kitamura Satoshi , Kawai Tadashi , Ohkubo Akiyuki , Hasegawa Shizuo , Kuriyama Takayuki , Kato Harubumi , Fukuoka Masahiro , Ohkawa Jiro
Published: August 2005

We evaluated the diagnostic usefulness of measurement of the soluble cytokeratin 19 fragment, a new tumor marker, in 391 patients with lung cancer and in 424 patients with benign lung diseases. Serum concentrations of cytokeratin 19 fragment were measured by a sandwich ELISA (CYFRA). The cut‐off value was defined as 3.5 ng/ml, which is associated with a specificity of 85% for benign lung diseases. CYFRA had a high sensitivity (57.5%) in all subjects with lung carcinoma, and had a higher sensitivity for squamous cell carcinoma (73.1%, n = 141) than squamous cell carcinoma‐related antigen (61.0%). CYFRA was associated with a relatively high sensitivity (42.1%) in early‐stage squamous cell carcinoma (stage I, based on the classification of the Japan Lung Cancer Society), but the CYFRA titer was higher in advanced squamous cell carcinoma than in early‐stage squamous cell carcinoma. Our findings suggest that CYFRA is potentially useful for diagnosis and monitoring of lung carcinoma, especially for squamous cell carcinoma.
Chemopreventive Effects of β‐Carotene, α‐Tocopherol and Five Naturally Occurring Antioxidants on Initiation of Hepatocarcinogenesis by 2‐Amino‐3‐methylimidazo[4,5‐f] qumoline in the Rat
Authors: Tsuda Hiroyuki , Uehara Nobuaki , Iwahori Yoshio , Asamoto Makoto , Ligo Masaaki , Nagao Minako , Matsumoto Kazuyuki , Ito Mitsuya , Hirono Iwao
Published: August 2005

Inhibitory effects of naturally occurring antioxidants on the initiation stage of hepatocarcinogenesis were studied. Group 1 rats were given a diet containing β‐carotene (β8‐CT, 0.02%), α‐tocopherol (α‐TP, 1.5%), glutathione (GLT, 5%), vanillin (VNL, 1%), quercetin (QCT, 1%) or ellagic acid (ELA, 1%), or 3 doses of diallyl sulfide (DAS, 200 mg/kg, i.g) over an 8‐day period. On day 7, the animals received a single dose of 2‐amino‐3‐methylimidazo[4,5‐f]jumoline (IQ, 100 mg/kg, i.g.), 12 h after two‐thirds partial hepatectomy for initiation and 2 weeks thereafter, were placed on promotion regimen comprising phenobarbital (0.05% in diet) and a single dose of D‐galactosamine (100 mg/kg, i.p.). Groups 2 and 3 were treated as described for Group 1, but without test material or IQ, respectively. Survivors were killed at week 11 and antioxidant influence was assessed by comparing values for preneoplastic glutathione S‐transferase placental form‐positive (GST‐P+) foci between Groups 1 and 2. All lesions larger than 70 βm in diameter consisting of approximately 5 cells in cross section were counted. Numbers of GST‐P+ foci/cm2 in Group 1 were: β ‐CT, 7.99; α‐TP, 8.21; GLT, 9.71; DAS, 10.37; VNL, 10.57; QCT, 11.1; ELA, 12.5 (n = 11‐15). All, except ELA, showed a significant decrease as compared with the Group 2 value of 14.54 (n=15). Only β ‐CT showed a significant decrease for the area value. This is the first report to show that β ‐CT, α‐TP, GLT, DAS, VNL, QCT exert inhibitory effects on initiation of hepatocarcinogenesis by the food carcinogen IQ, suggesting that these antioxidants might find application as chemopreventive agents. Furthermore, the current protocol proved practical for the assessment of chemopreventive agents within 11 weeks, a relatively Short period.
Rearrangement of bcl‐2 Is Detectable in Hodgkin's Disease by Polymerase Chain Reaction
Authors: Mitani Shōko , Oka Teruaki , Aoki Naoto , Hōjō Isao , ōta Ukiko , Mori Shigeo
Published: August 2005

The authors examined the occurrence of the t(14;18) chromosomal translocation in 44 cases of Hodgkin's disease (HD) using the polymerase chain reaction and Southern blot hybridization with non‐radioactive oligonucleotide probes. DNAs were extracted from unfixed, fresh‐frozen and formalin‐fixed, paraffin‐embedded biopsy specimens. Southern blot hybridization of the amplification product showed that, of 44 HD DNAs, three had a detectable t(14;18) breakpoint at the mbr (major breakpoint region), while none had a detectable t(14;18) breakpoint at the mcr (minor cluster region). Of the three cases positive for a t(14;18) breakpoint at the mbr, two were of lymphocyte predominance type, and the other was of mixed cellularity type.
Subtype Analysis of HTLV‐1 in Patients with HTLV‐1 Uveitis
Authors: Ono Ayako , Miura Tomoyuki , Araki Shinji , Yamaguchi Kazunari , Takatsuki Kiyoshi , Mori Sigeo , Hayami Masanori , Mochizuki Manabu , Watanabe Toshiki
Published: August 2005

The hypothesis that HTLV‐1 uveitis, a recently identified disease entity associated with human T‐cell leukemia virus type I (HTLV‐1), is caused by a specific subtype of the virus was tested. The nucleotide sequences of the long terminal repeat of HTLV‐1 from five patients with HTLV‐1 uveitis (HU) and four with adult T‐cell leukemia were phylogenetically analyzed. Our results showed that both subtypes which had been identified in Japan were associated with HU, indicating that there was no difference in pathogenicity between these phylogenetic subtypes. One of the subtypes was more frequently isolated in Okinawa than in Kyushu, suggesting a bias in the prevalence of each subtype among the inhabitants of these two areas of Japan.
Site‐specific Mutation of the Human c‐Ha‐ras Transgene Induced by Dimethylbenzanthracene Causes Tissue‐specific Tumors in Mice
Authors: Doi Satoru‐Takahiro , Kimura Minoru , Katsuki Motoya
Published: August 2005

Forestoraach squamous cell carcinomas, lung adenocarcinomas and spleen angiosarcomas were induced by dimethylbenzanthracene (DMBA) in the rasH2 transgenic mouse line carrying human c‐Ha‐ras genes with their own promoter, encoding the prototype p21 gene product. Fifteen out of 21 mice (71%) developed forestomach squamous cell carcinomas, while 15 out of 21 (71%) had lung adenocarcinomas and 3 out of 21 (14%) showed spleen angiosarcomas within 8 weeks after a single administration of 50 mg/kg DMBA intraperitoneally. Somatic mutation at the 61st codon of the transgenes, from CAG(Gln) to CTG(Len), was detected in all these newly developed tumors. However, non‐transgenic littermates demonstrated no tumors at all. These findings provide strong evidence that the somatic mutational activation of human c‐Ha‐ras genes is a critical event in tumorigenesis and a close relationship is therefore strongly suggested between the tissue‐specific development of tumors and the somatic mutation of human c‐Ha‐ras genes in these rasH2 transgenic mice.
Gastrin Receptor Gene Expression in Several Human Carcinomas
Authors: Matsushima Yumi , Kinoshita Yoshikazu , Nakata Hirohisa , Inomoto‐Naribayashi Yoko , Asahara Masakyo , Kawanami Chiharu , Nakamura Akira , Ito Mitsuhiro , Matsui Toshimitsu , Fujiwara Takato , Watanabe Hidenobu , Chiba Tsutomu
Published: August 2005

Gastrin has been shown to enhance the growth of various human tumors. The present study was designed to examine the gastrin receptor gene expression in various human carcinoma cell lines and in surgically resected carcinoma tissues. By Northern blot analysis, gastrin receptor mRNA was detected in 3 out of 7 small cell lung carcinoma cell lines. Gastrin receptor mRNA was also expressed in one out of 8 colon carcinoma cell lines and 2 out of 10 colon carcinoma tissues. Moreover, one of two small cell carcinoma cell lines of the stomach clearly expressed gastrin receptor mRNA. However, none of the gastric adenocarcinoma cell lines or surgically resected gastric adenocarcinomas tested had any detectable expression of gastrin receptor gene. These findings may suggest a role of gastrin receptor in the growth and differentiation of certain human carcinomas.
Nuclear Accumulation of p53 Protein Correlates with Mutations in the p53 Gene on Archival Paraffin‐embedded Tissues of Human Breast Cancer
Authors: Umekita Yoshihisa , Kobayashi Keiko , Saheki Takeyori , Yoshida Hiroki
Published: August 2005

Fifty invasive ductal carcinomas of the breast were analyzed by immunohistochemistry, polymerase chain reaction‐single strand conformationl polymorphism (PCR‐SSCP) and direct sequencing after microdissection of conventional formalin‐fixed, paraffin‐embedded tissues. A highly significant association between the presence of p53 gene mutation and nuclear accumulation of p53 protein was found (P< 0,0001). Of 13 tumors that demonstrated p53 gene mutations, 11 (84.6%) showed nuclear accumulation of p53 protein. However, of 37 tumors in which gene mutations were not detected, only 5 (13.5%) showed nuclear accumulation of p53 protein. There was a statistically significant association between the nuclear accumulation of p53 protein and a higher histological grade (P< 0.001) or mitotic index (P<0.01). In addition, gene mutations had a statistically significant association with a higher histological grade (P<0.05) or mitotic index (P> 0.0001). Therefore, p53 abnormalities might be associated with an aggressive phenotype in breast cancer. We conclude that the immunohistochem‐ical detection of nuclear p53 protein accumulation is highly associated with p53 gene mutations in archival paraffin‐embedded tissues, and that this method is useful for rapid screening of p53 abnormalities.
Transformation‐associated 86 kDa Natural Killer Target Molecule Expressed on the Mouse, Rat and Human Cell Surface
Authors: Qi Weimin , Tamura Yasuaki , Takashima Satoru , Sato Noriyuki , Kikuchi Kokichi
Published: August 2005

We previously reported on the 86 kDa natural killer target molecule associated with transformation of the oncogene‐transfected rat fibroblasts. This molecule may participate in the lethal hit phase of cytotoxicity by natural killer (NK) cells. Originally, this molecule was defined by mAb109, hut mAb109 could react only with rat tumor lines. In this report, to determine whether the 86 kDa molecule could be utilized as a natural killer target molecule in mammalian cells, we developed a polyclonal anti‐86 kDa antibody (pAh109). Our data indicated that pAh109 preferentially reacted with NK‐susceptible lines such as mouse YAC‐1, rat W31 and human K562 cells, but reacted only weakly with NK‐resistant mouse EL‐4, rat fetal fibroblast WFB and human fetal fibroblast HEPM. In a cytotoxicity experiment, pAb109 F(ab')2 fragments could inhibit the cytolysis by NK cells of W31 and K562 cells. However, these fragments did not inhibit the cytotoxicity of non‐NK cells such as CD3+, CD4−, CD8− T cell receptor αβ− ‐T cells (presumably γδ T cells) to W31 cells. Taken together, these data suggest that the cell transformation‐associated 86 kDa molecule may be critical in NK cytotoxicity, and a candidate for the NK target structure in mammalian tumor cells.
Detection of HTLV‐II‐seropositive Blood Donors in South Vietnam but Not in North Vietnam
Authors: Yamamoto Shinji , Nakata Susumu , Nakasone Tadashi , Chosa Tohru , Song Pham , Due Dao Dinh , Hien Bui , Quang Nguyen Xuan , Trinh Thi Ngoc , Ono Tomomichi , Honda Mitsuo
Published: August 2005

Approximately 1% (4/500) of blood donors exhibited seropositivity for HTLV‐II in South Vietnam, but none (0/500) did in North Vietnam. Further, all individuals seropositive for HTLV‐II were intravenous drug abusers who were seronegative for HIV‐1 and HTLV‐I. These findings suggest that HTLV‐II infection may be specifically prevalent in drug abusers in South Vietnam.
Germ Line Mutations of the ret Proto‐oncogene in Japanese Patients with Multiple Endocrine Neoplasia Type 2A and Type 2B
Authors: Maruyama Shoichi , Iwashita Toshihide , Imai Tsuneo , Funahashi Hiroomi , Ceccherini Isabella , Luo Yin , Romeo Giovanni , Matsuo Seiichi , Matsuyama Mutsushi , Takahashi Masahide
Published: August 2005

We investigated mutations of the ret proto‐oncogene in Japanese patients with multiple endocrine neoplasia (MEN) type 2A and type 2B. DNAs from pheochromocytomas and/or medullary thyroid carcinomas (MTCs) of five MEN 2A and three MEN 2B patients were amplified by a polymerase chain reaction (PCR) and analyzed. Tumors of four MEN 2A patients had missense mutations in Cys 634 in the extracellular domain of the ret proto‐oncogene. The same mutations were detected in normal tissues of the patients, indicating that the mutations had arisen in the germ line. Using a reverse transcriptase(RT)‐PCR, both normal and mutant transcripts of the ret proto‐oncogene were detected in a tumor of one patient with MEN 2A mutation. In addition, three MEN 2B patients examined had the same point mutation (ATGÂG) at codon 918 in the tyrosine kinase domain of the ret proto‐oncogene. Since all mutations identified in this study generated new restriction enzyme sites or eliminated a restriction site, the mutant alleles of affected family members could be readily detected without sequencing.
Role of Orthotopic Fibroblasts in the Development of Scirrhous Gastric Carcinoma
Authors: Yashiro Masakazu , Chung Yong‐Suk , Sowa Michio
Published: August 2005

We examined the interaction between scirrhous gastric cancer cells and organ‐specific fibroblasts in vivo and in vitro. Co‐inoculation of scirrhous gastric cancer cells with gastric fibroblasts into nude mice specifically increased tumorigenicity, compared with that of gastric cancer cells alone. Furthermore, the histologic findings of the xenograft produced by co‐inoculation with gastric fibroblasts was similar to that of human scirrhous gastric carcinoma. Conditioned medium from gastric fibroblasts significantly stimulated the growth of gastric cancer cells. These findings suggest that the growth of scirrhous gastric cancer cells was affected by orthotopic fibroblasts.
Apoptotic Cell Death in Human Gastric Carcinoma: Analysis by Terminal Deoxynucleotidyl Transferase‐mediated dUTP‐biotin Nick End Labeling
Authors: Kasagi Noriko , Gomyo Yoshihito , Shirai Hiroyuki , Tsujitani Shunichi , Ito Hisao
Published: August 2005

We have examined the occurrence of apoptotic cell death in formalin‐fixed, paraffin‐embedded human gastric carcinoma specimens by the terminal deoxynucleotidyl transferase (TdT)‐mediated dUTP‐biotin nick end labeling (TUNEL) method. The specificity of the TUNEL signals was confirmed by the omission of either TdT or biotinylated dUTP as negative controls, and by pretreatment with DNase I as a positive control. Careful observation of routine hematoxylin and eosin‐stained sections showed a few tumor cells with apoptosis, especially in well‐differentiated carcinomas. Intense TUNEL signals were frequently observed even in ordinary, non‐pyknotic nuclei of tumor cells, and occasionally also in nuclear fragments corresponding to apoptotic bodies. Apoptotic indices (number of apoptotic cells/total number of tumor cells) ranged between 7.7 and 14.5% (mean, 10.9%) in nine well‐differentiated carcinomas and between 2.7 and 7.5% (mean, 4.0%) in five which were poorly differentiated, the mean number being significantly higher in the former (P<0.01). No apparent correlation was found between apoptosis and the expression of proliferating cell nuclear antigen, P53 or Ley in the present study. This high frequency of apoptosis, implying cell loss, may be related to the slow‐growing nature of well‐differentiated carcinomas. Poorly differentiated carcinomas, including scirrhous gastric carcinomas, showed a lower incidence of apoptosis, indicating the existence of an escape mechanism from the process.
Inhibition of Tumor Necrosis Factor‐α and β Secretion by Lymphokine Activated Killer Cells by Transforming Growth Factor‐β
Authors: Naganuma Hirofumi , Sasaki Atsushi , Satoh Eiji , Nagasaka Mitsuyasu , Nakano Shin , Isoe Shiro , Tasaka Kachio , Nukui Hideaki
Published: August 2005

Transforming growth factor‐β (TGF‐β) has a variety of immunosuppressive properties. We investigated the effect of TGF‐β secreted by glioblastoma (T98G) cells on the secretion of tumor necrosis factor‐α and ‐β (TNFs) by lymphokine activated killer (LAK) cells stimulated with tumor cells. The supernatant from T98G cells was preincubated with anti‐TGF‐βl and ‐β2 neutralizing antibodies or untreated, and added to a coculture of LAK and Daudi cells. The neutralizing antibodies were added to LAK/Daudi and LAK culture, and natural human TGF‐β and recombinant human TGF‐β were also added to the LAK/Daudi culture. LAK cells were also cultured with T98G cells, of which the supernatant contained both active and latent forms of TGF‐/β1 and TGF‐β2, and the neutralizing antibodies were added to the coculture. TNFs activity in the supernatants from LAK/ Daudi cultures was examined by a specific bioassay. Addition of the supernatant from T98G cells to LAK/Daudi culture resulted in the inhibition of TNFs secretion by LAK cells. The inhibition was abrogated by the pretreatment of the supernatants with the anti‐TGF‐β antibodies. Addition of TGF‐β and TGF‐β to LAK/Daudi culture inhibited TNFs secretion by LAK cells in a dose‐dependent manner. Addition of anti‐TGFβ‐ antibodies to LAK culture resulted in an increase of TNFs secretion. These results suggest that, if tumor cells have the capacity to convert TGF‐β from a latent to an active form, the active TGF‐β suppresses TNFs secretion by LAK cells stimulated with the tumor cells, and that TGF‐β secreted and activated by glioblastoma cells suppresses the propagation of immune reaction by inhibiting TNFs secretion by activated lymphocytes adjacent to tumor cells.
Ovarian Teratomas in Mice Lacking the Protooncogene c‐mos
Authors: Furuta Yasuhide , Shigetani Yasuyo , Takeda Naoki , Iwasaki Kuniko , Ikawa Yoji , Aizawa Shinichi
Published: August 2005

Parthenogenesis has been suggested to be tightly coupled with development of ovarian teratomas. Indeed, ovarian tumors developed in c‐mos‐delieicnt female mice, which are characterized by the parthenogenetic activation of oocytes. The tumors appeared at a frequency of 30% between 4 and 8 months of age, and did not develop in younger or older mice. Most of the tumors were benign and consisted of multi‐focal cysts most notably with mature ectodermal components, but also with mesodermal and endodermal components. One among 17 tumors observed consisted of extraembryonic tissues alone, and two bore malignant components with metastasis to peritoneal organs. The results strongly suggest the involvement of c‐mos mutations in human germ cell tumors.
Frequent Amplification of the Cyclin E Gene in Human Gastric Carcinomas
Authors: Akama Yoshihiko , Yasui Wataru , Yokozaki Hiroshi , Kuniyasu Hiroki , Kitahara Kenji , Ishikawa Takenori , Tahara Eiichi
Published: August 2005

We searched for genetic alterations of the cyclin D1 and cyclin E genes in 45 human gastric carcinoma tissues. Expression of cyclin E mRNA and protein was also analyzed in eight of them by Northern and Western blots and immunohistochemical staining. The cyclin E gene was amplified 3–10 fold in seven gastric cancer tissues (15.6%), of which six were advanced gastric cancers. All of the cases with the cyclin E gene amplification displayed lymph node metastasis. Moreover, the case with the gene amplification overexpressed the cyclin E mRNA and protein. One of eight gastric cancer cell lines, MKN‐7, shared the cyclin E gene amplification, and all of the gastric cancer cell lines expressed high levels of the cyclin E mRNA and protein even without gene amplification. Amplification of the cyclin D1 gene was not observed in any of the gastric carcinoma tissues or gastric carcinoma cell lines. These results suggest that the gene amplification and overexpression of cyclin E play an important role in the abnormal growth and progression of gastric carcinoma.
Elevated Serum Growth Hormone Accelerates Gastric Tumorigenesis in F344 Rats after Treatment with N‐Methyl‐N‐nitrosourea in Their Drinking Water
Authors: Watanabe Hiromitsu , Fujimoto Nariaki , Kawamoto Keiichi , Ando Yasumi , Yamada Kazumasa , Okaraoto Taro , Kanin Georgie N. , Ito Akihiro
Published: August 2005

We examined the effects of growth hormone on tumorigenesis in F344 rats treated with N‐methyl‐N‐nitrosourea (MNU). Four‐week‐old male F344 rats were exposed to 100 ppm MNU in their drinking water for 15 weeks. Thereafter Group II animals received 100 μCi/100 g body weight of 131I (radiothyroidectomy, Tx) injected i.p. and Group III rats were implanted with pituitary tumors (MtT) secreting growth hormone while Group I received no further treatment after MNU. Non‐carcinogen control animals received MtT, Tx or no treatment. Animals were killed at 39 weeks after starting MNU administration. Gastric tumors were present in 13 of 31 (43%), 15 of 32 (47%) and 17 of 32 (53%) rats in Groups I to III, respectively. All tumors were of well‐differentiated type. Spinal cord tumors appeared in 15 of 31 (47%) in Group I, 10 of 32 (32%) in Group II and 10 of 32 (32%) in Group III, most being malignant schwannomas. Thymic lymphornas also appeared in 10 of 31 (32%), 5 of 32 (16%) and 6 of 32 (19%) animals in Groups I to III, respectively. There were no significant differences among the groups. However, tumors in Group III developed significantly earlier than in Groups I or II. This was mainly due to gastric tumors, and cumulative incidence curves for spinal cord tumors or thymic lymphomas were similar in all groups. The results indicate that gastric tumors induced by MNU in F344 male rats are influenced by elevated levels of growth hormone.
Increased Midkine Gene Expression in Human Gastrointestinal Cancers
Authors: Aridome Kuniaki , Tsutsui Jun‐ichiro , Takao Sonshin , Kadomatsu Kenji , Ozawa Masayuki , Aikou Takashi , Muramatsu Takashi
Published: August 2005

Midkine (MK) is a product of a retinoic acid‐responsive gene, and is a novel growth differentiation factor. We examined the expression of the MK gene in specimens of 47 surgically removed human carcinomas of the gastrointestinal organs, namely, gastric, colorectal, hepatocellular, pancreatic, esophageal, ampullary duodenal and bile duct carcinomas. In most cases, the MK mRNA level was higher in cancer specimens than in the corresponding non‐cancerous tissues. Furthermore, MK mRNA was more highly expressed in the colon adenocarcinoma lesion than in the adenoma lesions, in the two familial polyposis cases. While MK mRNA was not detected in the normal liver, it became detectable in cirrhotic tissues in 2 of 4 cases, and its expression was increased in the cancerous tissues. Thus, the increase of MK mRNA level is a phenomenon seen in many human gastrointestinal carcinomas. The increased expression of the MK gene in gastric carcinoma was significantly more prominent in well and moderately differentiated adenocarcinomas than in poorly differentiated adenocarcinomas and signet ring cell carcinomas.
Induction of Invasive Growth in a Gallbladder Cancer Cell Line by Hepatocyte Growth Factor in vitro
Authors: Shimura Hideo , Date Kazuhiko , Matsumoto Kunio , Nakamura Toshikazu , Tanaka Masao
Published: August 2005

To study the mechanism of invasion and metastasis of gallbladder cancer cells, we established a cancer cell line, GB‐d1, from a metastatic lymphnode of poorly differentiated adenocarcinoma of the gallbladder. GB‐d1 cells proliferate well in a dish culture and form small cystic cell clusters in a collagen gel containing 10% fetal bovine serum. A conditioned medium of human embryonic lung fibroblasts (HEL) stimulated the proliferation of GB‐d1 cells and induced cell scattering in the dish culture. In the gel culture, the conditioned medium induced a transformation of the spherical clusters to arborizating colonies with tubular projections that mimicked an invasion of cancer cells into the surrounding tissue. Similar results were obtained when 10 ng/ml of human recombinant hepatocyte growth factor (h‐rHGF) was added to the culture medium. The proliferative and morphological changes induced by the conditioned medium were inhibited by antiserum against h‐rHGF. HEL and human gallbladder stromal fibroblast‐like cells produced substantial levels of HGF in the culture media, while GB‐d1 did not produce any detectable level of HGF. These results suggest that HGF promotes the invasive growth of gallbladder cancer cells in vitro, and it was also suggested that stromal fibroblasts may play an important role in the invasive progression of gallbladder cancer in a paracrine fashion.
Stimulatory Effect of Interleukin‐1α on Proliferation through a Ca2+/Calmodulindependent Pathway of a Human Thyroid Carcinoma Cell Line, NIM 1
Authors: Inokuchi Nobuo , Zeki Kazuya , Morimoto Isao , Nakano Yoichiro , Fujihira Takashi , Yamashita Uki , Yanagihara Nobuyuki , Izumi Futoshi , Eto Sumiya
Published: August 2005

NIM 1 cells, a human thyroid cell line established from a patient with thyroid papillary adenocarcinoma, produce cytokines such as interleukin‐1α (IL‐1α) and granulocyte‐colony stimulating factor. In the present study, we investigated the signal transduction pathway in the proliferation of NIM 1 cells evoked by IL‐1α. Incubation of NIM 1 cells with IL‐1α for 48 h increased the incorporation of 3H‐thymidine (3H‐TdR). The stimulatory effect of IL‐1α was evident at 0.01 ng/ml and the maximal effect was seen at 10 ng/ml. IL‐1α evoked an influx of 45Ca into NIM 1 cells within 3 min in a concentration‐dependent manner (0.01–1 ng/ml). These stimulatory effects of IL‐1α on both 3H‐TdR incorporation and 45Ca influx were similarly inhibited by nicardipine, an inhibitor of voltage‐dependent Ca2+ channels, in a concentration‐dependent manner (10–1000 nM). The stimulatory effect of IL‐1α on 3H‐TdR incorporation was inhibited by N‐(6‐aminohexyl)‐5‐chloro‐1‐naphthalenesulfonamide (W‐7), an antagonist of calmodulin, but not by 1‐(5‐isoquinoline sulfonyl)‐2‐methylpiperazine (H‐7), an inhibitor of protein kinase C. While the culture medium initially contained 0.75 mM Ca2+, inhibition of 3H‐TdR incorporation by nicardipine and W‐7 under these baseline conditions was also recognized. These results suggest that IL‐1α stimulates cell proliferation through a Ca2+/calmodulin‐dependent pathway in NIM 1 cells.
Inhibitory Effects of Medroxyprogesterone Acetate on Mouse Endometrial Carcinogenesis
Authors: Niwa Kenji , Morishita Shigeo , Murase Toshiko , Itoh Naoki , Tanaka Takuji , Mori Hideki , Tamaya Teruhiko
Published: August 2005

The present study was undertaken to examine the effects of cyclic administration of low‐dose progestogen on endometrial carcinogenesis in mice. A total of 115 female ICR mice, 10 weeks of age, were divided into four experimental and control groups. Mice in groups 1–3 received laparotomy and were injected with N‐methyl‐N‐nitrosourea (MNU) solution at a dose of 1 mg/100 g body weight to the left uterine tube and with normal saline to the right uterine tube. From one week after the MNU exposure, groups 1 and 2 were given 5 ppm 17β‐estradiol (E2)‐containing diet throughout the experiment. Mice in group 1 received 5 s.c. injections of medroxyprogesterone acetate (MPA) (2 mg/ mouse) at intervals of 4 weeks from week 7. Group 3 was treated with MNU/normal saline alone. Group 4 consisted of mice treated with MPA alone. At the termination of the experiment (week 30), all animals were killed and autopsied for pathological examinations. It was found that adenocarcinomas and preneoplastic lesions developed in the bilateral uterine corpora in mice of groups 1–3. MPA treatment significantly decreased the weight of the uterine corpus (P< 0.05) and the incidences of endometrial adenocarcinoma and atypical or adenomatous (P< 0.001) but not cystic glandular hyperplasias in the MNU/E2‐treated groups. Additionally, MPA treatment tended to decrease the proliferating cell nuclear antigen‐labeling index in endometrial glandular cells. These data indicate that MPA, even at low dose, has an inhibitory effect on mouse endometrial carcinogenesis induced by MNU and E2.
Combination Assay of Urinary β‐Core Fragment of Human Chorionic Gonadotropin with Serum Tumor Markers in Gynecologic Cancers
Authors: Kinugasa Masato , Nishimura Ryuichiro , Koizumi Tamio , Morisue Kouichi , Higashida Taro , Natazuka Toshiki , Nakagawa Toshitaro , Isobe Takashi , Baba Shigeaki , Hasegawa Kazuo
Published: August 2005

Ectopic production of the immunoreactive β‐subunit of human chorionic gonadotropin (IR‐hCGβ) by gynecologic malignancies has been well recognized, but IR‐hCGβ has not yet been established as a clinically useful tumor marker, except for germ cell tumors. We measured the concentrations of IR‐hCGβ‐related molecules, intact hCG, free hCGβ, and β‐CF, in the sera and urine of patients with various gynecologic cancers (cervical, endometrial, and ovarian cancers) to assess their clinical usefulness as a tumor marker in comparison with serum tumor markers such as CEA, SCC, CA125, and CA19‐9. The highest incidence of IR‐hCGβ was obtained in tbe assay for β‐CF in the urine, with positive rates of 47.7% (94 of 197) for cervical, 37.8% (14 of 37) for endometrial, and 84.4% (38 of 45) for ovarian cancers with a cut‐off value of 0.2 ng/mg of creatinine. In cervical cancer, there was no significant correlation between the concentrations of urinary β‐CF and serum SCC, and 57.9% (114 of 197) of the patients were detected by the combination assay of these tumor markers. Serial determination in 22 cervical cancer patients with elevated urinary β‐CF level prior to therapy showed that its level decreased after successful treatment, but 4 of 5 patients with persistent or recurrent disease had elevated levels of urinary β‐CF. All of the ovarian cancer patients examined were detected by the combination assay of urinary β‐CF and serum CA125. The levels of urinary β‐CF showed little correlation with those of the serum tumor markers, indicating the usefulness of the combination assay of urinary β‐CF with serum tumor markers for detecting cervical and ovarian cancers.
Genetic Complementation of the Immortal Phenotype in Group D Cell Lines by Introduction of Chromosome 7
Authors: Ogata Toshihiko , Oshimura Mitsuo , Namba Masayoshi , Fujii Michihiko , Oishi Michio , Ayusawa Dai
Published: August 2005

Human immortal cell lines have been classified into at least four (A–D) genetic complementation groups by cell‐cell hybrid analysis, i.e., a hybrid derived from different groups becomes mortal. Recently we have demonstrated that introduction of human chromosome 7 suppresses indefinite division potential in the non‐tumorigenic human immortalized fibroblast lines KMST‐6 and SUSM‐1, both assigned to complementation group D. By extending our microcell‐mediated chromosome transfer, we found that chromosome 7 also suppresses division potential in the human hepatoma line HepG2 (again, assigned to group D). Chromosome 7 was thus shown to suppress indefinite growth in the above group D cell lines irrespective of their cell types, or whether they are tumorigenic or not. Since chromosome 7 had no such effect on representative cell lines derived from complementation group A, B or C, these results indicate that the senescence gene(s) commonly mutated in the group D cell lines is located on chromosome 7.
Human Alveolar Macrophages Augment Natural Killer Cell Stimulatory Factor (Interleukin‐12)‐inducible Killer Activity from Autologous Blood Lymphocytes
Authors: Haku Takashi , Sone Saburo , Nabioullin Roustem , Ogura Takeshi
Published: August 2005

Interleukin‐12 (IL‐12), also known as natural killer cell stimulatory factor (NKSF), was found to induce cytotoxic activity from human blood T cells and NK cells. The present study was undertaken to examine the effect of human alveolar macrophages (AM) on induction by IL‐12 cytotoxic cells from blood lymphocytes. AM were obtained by bronchoalveolar lavage from healthy donors. Highly purified lymphocytes (>99%) and monocytes (>90%) were also isolated by centrifugal elutriation from peripheral blood of the same donors. Cytotoxicity of lymphocytes was measured by 4‐h 51Cr release assay. IL‐12 stimulated blood lymphocytes to produce interferon γ (IFNγ) and tumor necrosis factor α (TNFα), and this effect was augmented by co‐cultivation with monocytes or AM. AM‐upregulated induction of cytotoxic lymphocytes was stimulated with IL‐12, and this effect was significantly abrogated by addition of antibodies against IFNγ and TNFα. Induction by IL‐12 of IFNγ production and cytotoxic activity of CD8+ cells was also augmented by co‐cultivation with monocytes or AM. AM were more effective than monocytes in augmenting the cytotoxic activity of IL‐12‐stimulated lymphocytes and CD8+ cells. These observations suggest that in situ induction of IL‐12‐stimulated cytotoxic cells in the lung may be regulated by complex cytokine networks, depending on participation of monocytes and alveolar macrophages.
Intracellular Carboxyl Esterase Activity Is a Determinant of Cellular Sensitivity to the Antineoplastic Agent KW‐2189 in Cell Lines Resistant to Cisplatin and CPT‐11
Authors: Ogasawara Hayato , Nishio Kazuto , Kanzawa Fumihiko , Lee Yong‐Sik , Funayama Yasunori , Ohira Tatsuo , Kuraishi Yasunobu , Isogai Yukihide , Saijo Nagahiro
Published: August 2005

KW‐2189, a novel antitumor antibiotic belonging to the duocarmycins, possesses marked DNA‐binding activity upon activation by carboxyl esterase to its active form, DU‐86. Three duocarmycins, KW‐2189, DU‐86 and duocarmycin SA, were active against the cisplatin (CDDP)‐resistant human non‐small cell lung cancer cell lines PC‐9/CDDP and PC‐14/CDDP, and the multidrug‐resistant human small cell lung cancer cell line H69/VP. However, HAC2/0.1, a CDDP‐resistant human ovarian cancer cell line which is also resistant to CPT‐11 because of decreased intracellular activation of CPT‐11, was about 12.8‐fold more resistant to KW‐2189. HAC2/0.1 was not resistant to other duocarmycins as compared to its parental cell line, HAC2. There was no difference between HAC2 and HAC2/0.1 with regard to the intracellular accumulation of KW‐2189. Addition of 130 mU/ml of carboxyl esterase to the culture medium did not influence the sensitivity of HAC2 cells to KW‐2189. However, the sensitivity of HAC2/0.1 cells to KW‐2189 was enhanced to the level of HAC2. These results suggest that HAC2/0.1 is less potent than HAC2 in activating KW‐2189. The carboxyl esterase activity of whole‐cell and microsomal extracts from HAC2/0.1 was approximately 60% of that from HAC2. The cell‐free experiment revealed that KW‐2189 bound to DNA more efficiently in the presence of HAC2 than HAC2/0.1 cell extract. It was concluded that decreased intracellular carboxyl esterase activity in HAC2/0.1 cells caused decreased intracellular conversion of KW‐2189 to its active form, thus producing resistance to KW‐2189. The decreased conversion of CPT‐11 to SN‐38 in HAC2/0.1 cells might be explained by decreased carboxyl esterase activity.
Effects of Menstrual and Reproductive Factors on the Risk of Breast Cancer: Meta‐analysis of the Case‐Control Studies in Japan
Authors: Nagata Chisato , Hu Yao‐Hua , Shimizu Hiroyuki
Published: August 2005

To elucidate the magnitude of the effect of menstrual and reproductive factors on breast cancer occurrence among Japanese women, we reviewed eight case‐control studies previously conducted in Japan and used a quantitative method (meta‐analysis) to summarize the data. While individual studies have different methods and populations, the estimated odds ratios (ORs) in the studies were statistically homogeneous for all study variables. It was confirmed that early age at menarche, late age at first birth, and premenopausal status are significantly associated with risk of breast cancer; an estimated combined OR of 0.68 (95% confidence interval (CI): 0.59‐0.77) was obtained for women with onset of menstruation after age 16 compared to those before age 14. Nulliparous women had higher risk than women with first birth before age 25 (OR=1.56 95%, CI: 1.27‐1.91). The OR for women with first birth after age 35 was 2.26 (95% CI: 1.85‐2.77) compared to women at first birth before age 25. Premenopausal women had a higher risk than women with menopause before age 50 (OR=2.21, 95% CI: 1.53‐3.20). We also found a significant protective effect of high parity after controlling for age at first birth and the other menstrual factors. The OR estimate for 3 or more births compared to nulliparity was 0.68 (95% CI: 0.54‐0.86). The meta‐analysis provided quantitative estimates of breast cancer risk among Japanese women with improved precision.
Low Incidence of a Nucleotide Sequence Alteration of the Neurofibromatosis 2 Gene in Human Breast Cancers
Authors: Yaegashi Sadanori , Sachse Richard , Ohuchi Noriaki , Mori Shozo , Sekiya Takao
Published: August 2005

We investigated aberrations of the neurofibromatosis type 2 (NF2) gene in breast tumors of 60 patients by single‐strand conformation polymorphism analysis of polymerase chain reaction products followed by nucleotide sequencing. We detected a tumor‐specific single‐base substitution in codon 398 in exon 12 of the gene, resulting in an alteration of a single amino acid, in DNA from a breast cancer sample. The results indicated possible but infrequent involvement of mutations of the tumor suppressor NF2 gene in human breast cancers.
Multidrug Resistance‐associated Protein‐mediated Multidrug Resistance Modulated by Cyclosporin A in a Human Bladder Cancer Cell Line
Authors: Kim Wun‐Jae , Kakehi Yoshiyuki , Hirai Midori , Arao Shinji , Hiai Hiroshi , Fukumoto Manabu , Yoshida Osamu
Published: August 2005

A doxorubicin‐resistant subline (5637/DR5.5) from human bladder cancer cells (5637) was induced by stepwise increase in the doxorubicin concentration. 5637/DR5.5 cells were cross‐resistant to vinblastine and etoposide but not to mitomycin C and cisplatin. We analyzed the mdr1, MRP (multidrug resistance‐associated protein), and DNA topoisomerase II gene expression using the reverse transcription polymerase chain reaction assay (RT‐PCR) and investigated possible differences in the accumulation and efflux of radiolabeled daunorubicin. 5637/DR5.5 cells do not express the mdr1 gene, but the expression levels of MRP are markedly higher than in drug‐sensitive 5637 cells. The intracellular accumulation of radiolabeled daunorubicin was markedly decreased in the 56377 DR5.5 cells in comparison with the parent cells. This reduced drug accumulation was associated with an enhanced drug efflux, but was reversed when cells were incubated with cyclosporin A. Cyclosporin A at the concentration of 5 (M caused 3.4‐fold enhancement of daunorubicin‐sensitivity in the 56377 DR5.5 cells. On the other hand, there was no difference in DNA‐topoisomerase II activity between the parent and resistant cells. The resistance of the 5637/DR5.5 cells is therefore associated with an enhanced drug efflux mediated by the MRP gene overexpression, as distinct from P‐glycoprotein, and is modulated by cyclosporin A.
Diagnosis and Treatment of Esophageal Neoplasms
Authors: Kato Hoichi
Published: August 2005

During the last 10 years, the diagnosis and treatment of esophageal carcinoma have improved considerably. Endoscopy with Lugol staining and endoscopic ultrasonography have been newly introduced and used for early diagnosis and more accurate tumor staging. As a result, the number of patients with tumors at an early stage has increased remarkably (superficial carcinoma, 23%). In the field of treatment, surgical results have improved not only in the short term (30‐day mortality rate, 4%) but also in the long term (5‐year survival rate, 30%). The field of operation has been extended (3‐fieId lymph node dissection), with lower morbidity and mortality. On the other hand, some techniques for limited treatment such as endoscopic mucosal resection, intraluminal radiotherapy, and laser irradiation have been introduced for the treatment of esophageal carcinoma at an early stage with curative intent. However, there are still many patients with esophageal carcinoma at an advanced stage for whom these treatments fail or are futile. The role of radiotherapy has been made more significant by the introduction of brachytherapy or in combination with other treatment modalities such as surgery, chemotherapy and hyperthermia. Response rates for existing anticancer drugs used as a single agent are 0‐38%. Chemotherapy appears to have created significant improvements when used in combined modalities (response rate, 16‐76%). However, chemotherapy for patients with esophageal carcinoma still offers an unsatisfactory survival benefit and remains experimental. Studies to evaluate multimodality treatments using chemotherapy, combined with radiotherapy and/or surgery have started. The contribution of molecular biology to the diagnosis and treatment of this disease is a subject for future investigation.
Infectious Retrovirus Is Inactivated by Serum but Not by Cerebrospinal Fluid or Fluid from Tumor Bed in Patients with Malignant Glioma
Authors: Shimizu Keiji , Miyao Yasuyoshi , Tamura Masakazu , Kishima Haruhiko , Ohkawa Motohisa , Mabuchi Eiichiro , Yamada Masanobu , Hayakawa Toru , Ikenaka Kazuhiro
Published: August 2005

Intravenous gene transfer using recombinant retroviruses tends to suffer from a low infectious viral titer when conducted in vivo. This is, in part, caused by complement‐mediated proteolytic inactivation of the retrovirus in human serum. However, if the retroviruses were directly injected into the brain, they might not be inactivated. Supernatant from amphotropic retrovirus‐producing cells harboring the BAG vectors was incubated with sera or cerebrospinal fluid (CSF) of patients with gliomas or unrelated disorders. The retroviruses were severely inactivated in sera. However, no such inactivation was noted in CSF or fluid from the tumor bed of glioma patients. These data suggest that gene transfer using recombinant retroviruses could be done into the cavity after removal of the tumor in glioma patients.
Effect of Hammerhead Ribozyme against Human Thymidylate Synthase on the Cytotoxicity of Thymidylate Synthase Inhibitors
Authors: Kobayashi Hiroyuki , Takemura Yuzuru , Miyachi Hayato , Skelton Lorraine , Jackman Ann L.
Published: August 2005

One of the resistance mechanisms to folate‐based thymidylate synthase (TS) inhibitors is the increase in TS activity in tumor cells. Human B lymphoblastoid cell line (W1L2) was made resistant to a lipophilic non‐polyglutamatable TS inhibitor (ZM249148), and the subline (W1L2:R179) showed a 20‐fold increase in TS enzyme activity with concomitant overexpression of TS mRNA. To overcome the resistance, we designed a ribozyme that can cleave the CUC sequences in a triple tandemly repeated sequence of TS mRNA. Expression of this ribozyme in W1L2:R179 cells transfected with Epstein Barr virus‐based expression vector resulted in sensitization to TS inhibitors concomitantly with a decrease of TS expression. The ribozyme expressed in transfectants was shown to be functional in cleaving artificial TS RNA in vitro.
Epithelial‐cadherin Gene Is Not Mutated in Ductal Carcinomas of the Breast
Authors: Kashiwaba Masahiro , Tamura Gen , Suzuki Yasushi , Maesawa Chihaya , Ogasawara Satoshi , Sakata Ken , Satodate Ryoichi
Published: August 2005

We investigated mutations of the epithelial (E)‐cadherin gene and loss of heterozygosity (LOH) at flanking loci using three microsatellite markers on the long arm of chromosome 16 in 25 ductal carcinomas of the breast. Expression of E‐cadherin was also investigated immunohistochemically. No mutations were detected in exons 6 through 9 of the E‐cadherin gene. LOH was observed more frequently (42%) at D16S402 (16q‐ter) than at D16S421 (16q22.3‐q23.1) (17%), which is located near the E‐cadherin gene. Expression of E‐cadherin was observed at the cell borders in 92% (11/12) of the tumors examined. The absence of mutations in the E‐cadherin gene and its conserved expression suggest that inactivation of E‐cadherin does not contribute significantly to the invasion or metastatic potential of ductal carcinomas of the breast. Furthermore, the high frequency of LOH at 16q‐ter suggests the existence of another tumor suppressor gene which may play a crucial role in the genesis of ductal carcinomas of the breast.