bluuebunny/crossref_metadata_2025 huggingface dataset:
Serum Levels of Laminin, Type IV Collagen and Type III Procollagen Peptide as Markers for Detection of Metastasis
Authors: Yudoh Kazuo , Matsui Hisao , Kanamori Masahiko , Ohmori Kazuo , Tsuji Haruo , Tatezaki Shinichiro
Published: August 2005

We investigated the clinical usefulness of serum laminin, type IV collagen and type III procollagen peptide (PIIIP) as markers for detection of metastasis in patients with primary or mctastatic hone and soft part tumors. The subjects consisted of 28 patients with metastatic bone tumors, 18 with primary bone tumors (benign; 10, malignant; 8), 22 with primary soft part tumors (benign; 12, malignant; 10), 18 with cancer without metastasis (as controls to metastatic bone tumor) and 60 healthy controls. Elevated levels of serum laminin, type IV collagen and PIIIP were not associated with any specific histological subtype, tumor size or location, and were clearly related to evidence of metastasis. Mean serum concentrations of laminin, type IV collagen and PIIIP were significantly higher in patients with metastasis than in patients without metastasis. Positive correlations were observed among serum laminin, type IV collagen and PIIIP levels in tumor patients. The sensitivity values for laminin, type IV collagen and PIIIP in detecting metastasis were 83.7%, 83.3% and 80.5%, respectively, with specificity of 90.0%, 86.1% and 86.1%. When two of the three markers were evaluated in identical blood samples, combined sensitivity and specificity values exhibited further increases as compared to the sensitivity and specificity of each marker. The use of all three markers led to the best combined sensitivity and specificity. These findings suggest that the combination of these markers would be a valuable screening test in predicting metastasis.
The WD Gene for Wilson's Disease Links to the Hepatitis of LEG Rats
Authors: Ono Takao , Takada Shigeyuki , Yoshida Michihiro C.
Published: August 2005

LEC rats develop an autosomal recessive hepatitis and subsequently liver cancer associated with copper accumulation in the liver similar to that of Wilson's disease. Using 71 backcross [(WKAH x LEC) x LEC] rats, linkage analysis of the hepatitis with the WD gene for Wilson's disease revealed identical segregation and no recombination event between these two genes. This result indicates that the WD gene is a prime candidate for the hts gene responsible for the hepatitis of LEG rats, and suggests that the hepatitis of LEC rats may be caused by a defect in a copper‐transporting ATPase expressed in the liver.
Variant Translocation of the BCL6 Gene to Immunoglobulin k Light Chain Gene in B‐Cell Lymphoma
Authors: Suzuki Ken , Miki Tohru , Kawamata Norihiko , Hirosawa Shinsaku , Yoshizawa Kaname , Kiyosawa Kendo , Aoki Nobuo
Published: August 2005

A lymphoma cell line with a variant type of translocation, t(2;3)(pll;q27), was established from a patient who had received liver transplantation. To elucidate the molecular mechanism of the t(2;3)(pll;q27) chromosomal translocation, we compared the structures of both derivative (der) chromosomal breakpoints with those of their germline predecessors. We noted that the BCL6 gene on chromosome 3 was juxtaposed with the immunoglobulin k light chain (Igk) gene on chromosome 2 in a head‐to‐head configuration. The breakpoint of the BCL6 gene was within a previously reported breakpoint cluster region. The breakpoint on chromosome 2 was within the intron between the leader (L) and variable (V) sequences of one of the Vk genes, which was fused to the Jk3 (J=joining) segment. At chromosomal junctures, a direct repeat duplication of chromosome 3 sequences and a deletion of chromosome 2 sequences were discovered. These results are consistent with a translocation model with illegitimate pairing of staggered double‐stranded DNA breaks at 3q27 and 2pll, repair, and ligation to generate der(3) and der(2) chromosomes.
SlOO‐positive Histiocytes in T‐Cell‐dependent Area in Human Lymph Nodes Express P‐Glycoprotein
Authors: Nishii Kazuhiro , Kita Kenkichi , Miwa Hiroshi , Ohno Toshiyuki , Yamaguchi Motoko , Oka Kohji , Shirakawa Shigeru , Fukumoto Manabu
Published: August 2005

In the present study, we examined P‐glycoprotein (P‐gp)‐expressing cells in human lymph nodes (LN) from patients with reactive lymph adenitis and various malignant lymphomas (ML). MDRl mRNA was detected in all reactive LN and in 8 of 10 ML samples examined hy reverse transcription polymerase chain reaction (RT‐PCR). The signal intensity of the bands was stronger in samples from T‐cell ML LN than in those from LN of reactive lymph adenitis and B‐celt ML. Immunohisto‐chemistry also showed positive reactivity with cellular components in the LN. The P‐gp‐positive cells, detected by UIC2, in reactive lymph adenitis and ML samples were large in size in the T‐cell‐dependent paracortical area in LN. Further, morphological characteristics were similar in UIC2‐positive cells between reactive and ML LN. Double staining with UIC2 and a hematopoietic cell lineage‐specific monoclonal antibody showed that UIC2‐positive cell co‐expressed S‐100, but not other lymphocyte‐ and ML cell‐specific antigens, indicating that these cells correspond to histiocytes in the T‐cell‐dependent paracortical area. Our results show that P‐gp was expressed in histiocytes but not ML cells in LN, and suggest that this molecule may play an important role in the biological function of histiocytes and/or for maintenance of homeostatic levels in T‐cell ML LN.
Synergistic Enhancement of Cisplatin Cytotoxicity by SN‐38, an Active Metabolite of CPT‐11, for Cisplatin‐resistant HeLa Cells
Authors: Minagawa Yukihisa , Kigawa Junzo , Ishihara Hiroshi , Itamochi Hiroaki , Terakawa Naoki
Published: August 2005

A cisplatin (cis‐diamininedichloroplatinuin(II); CDDP)‐resistant HeLa cell line (HeLa/CDDP cells), which showed more than 8‐fold resistance to CDDF compared to the parent cells, was newly established for this study. HeLa/CDDP cells accumulated 50% less platinum than the parent cells. There was no difference in intracellular glutathione (GSH) content between the parent and HeLa/ CDDP cells. The dose modification factor by DL‐buthionine‐S, R‐sulfoximine (BSO) pretreatment was similar in both cell lines. HeLa/CDDP cells had cross‐resistance to diammine (l, l‐cyclobutanedicarboxylato)platinum(II) (CBDCA), (cis‐diammine (glycolato)platinum (254‐S), but not to (‐)‐(R)‐2‐aminomethylpyrrolidine(1,1‐cyclobutanedicarboxylato)platinum(II) (DWA2114R), adriamycin, or VP‐16. HeLa/CDDP cells showed a collateral sensitivity to 7‐ethyl‐10‐hydroxycampto‐thecin (SN‐38), an active metabolite of 7‐ethyl‐10‐[4‐(l‐piperidino)‐l‐piperidino]carbonyloxycamptothecin (CPT‐11). Furthermore, isobologram analysis indicated synergistic interaction of CDDP and SN‐38 only for HeLa/CDDP cells. The present study suggests that combination therapy with CDDP and CPT‐11 may he potentially useful in the treatment of some patients with CDDP‐resistant cancer.
Point Mutations of ras and Gsα Subunit Genes in Thyroid Tumors
Authors: Horie Hidetaka , Yokogoshi Yutaka , Tsuyuguchi Masaru , Saito Shiro
Published: August 2005

We studied 43 thyroid tumors including 5 adenomatous goiters, 7 follicular adenomas, 22 papillary carcinomas, and 9 medullary carcinomas with regard to the presence of point mutations in the genes of Gs alpha subunit (Gsα), Gi2 alpha subunit (Gi2α), H‐ras, K‐ras, and N‐ras by a polymerase chain reaction‐direct sequencing method. An adenomatous goiter and a follicular adenoma showed double mutations at codon 227 and 231, and 4 papillary carcinomas showed mutation at codon 231 of the Gsα gene. An adenomatous goiter, a follicular adenoma, and a papillary carcinoma showed a missense mutation in codon 13 of the K‐ras gene. There were no such missense mutations of these G‐protein or ras genes in medullary carcinomas. These data indicate that the genetic events involved in the oncogenesis of parafollicular C‐cells are different from those of thyroid follicular cells, in which missense mutations of Gsα and ras genes seem to play important roles in tumorigenesis.
Microsatellite Instability and Other Molecular Abnormalities in Human Prostate Cancer
Authors: Suzuki Hiroyoshi , Komiya Akira , Aida Sara , Akimoto Susumu , Shiraishi Taizo , Yatani Ryuichi , Igarashi Tatsuo , Shimazaki Jun
Published: August 2005

Microsatellites are highly polymorphic, short‐tandem repeat sequences dispersed throughout the genome. Instability of these repeat sequences at multiple genetic loci may result from mismatch repair errors, and occurs in hereditary nonpolyposis colorectal carcinoma and certain sporadic cancers. To examine microsatellite instability during the pathogenesis of human prostate cancer, we screened 48 prostate cancer cases (20 stage B, 10 stage C and 18 endocrine therapy‐resistant cancer‐death cases) for replication error at 17 microsatellite marker loci on 9 chromosomes. Microsatellite instabilities were found in 7 of 48 cases (14.6%), and all 7 cases showing the instability were poorly differentiated adenocarcinomas. Moreover, microsatellite instabilities were more frequently observed in cancer‐death cases (6/18, 33%) than in stage B+C cases (1/30, 3.3%). These data suggest that micro‐satellite instability is an important genetic change related to the progression of a subset of human prostate cancer cases. It is suggested to be associated with extensive, concurrent molecular changes including androgen receptor gene mutations, as well as frequent loss of heterozygosity at chromosomal regions 8p, 10q, and 16q.
Biomodulation by Hyperthermia of Topoisomerase II‐Targeting Drugs in Human Colorectal Cancer Cells
Authors: Hirohashi Yoshimi , Hidaka Katsuhiko , Sato Seiji , Kuwano Michihiko , Kohno Kimitoshi , Hisatsugu Takeharu
Published: August 2005

We examined whether heat stress could enhance the sensitivity of human colon cancer WiDr cells to topoisomerase II‐targeting anticancer agents, etoposide (VP‐16) and teniposide (VM‐26), and also determined the most effective timing for the drug administration after exposure to hyperthermia. Both topoisomerase II contents and topoisomerase II activity were significantly increased in WiDr cells 3 to 12 h after heat stress at 43°C for 1 h, in comparison with those immediately after the heat stress. Cytotoxicity by VP‐16 was most significantly enhanced 3 to 12 h after exposure to 43°C for 1 h, but no synergistic effect was observed when the drug was administered immediately after the heat stress. A combination of VM‐26 with heat stress, but not that of a topoisomerase I‐targeting camptothecin derivative (CPT‐11), or vincristine, showed a synergistic cytotoxic effect on WiDr cells. VP‐16 alone induced cellular accumulation at the G2+M phase, whereas the combination of VP‐16 and heat stress further increased the cell population at the G2+M phase, and decreased S‐phase cells. A possible application of the combination of VP‐16 and hyperthermia in clinical use is discussed.
Inhibition by Differentiation‐inducing Agents of Wild‐type p53‐dependent Apoptosis in HL‐60 Cells
Authors: Noguchi Kohji , Nakajima Motowo , Naito Mikihiko , Tsuruo Takashi
Published: August 2005

The product of the p53 tumor‐suppressor gene has been shown to function in apoptosis and cell cycle regulation. However, there is little information regarding the regulation of apoptosis in cell differentiation. We investigated the relationship between p53‐dependent apoptosis and differentiation induction using human promyelocytic leukemia HL‐60 cells transfected with pMAMneo expression vectors containing dexamethasone‐inducible wild‐type p53 (wt‐p53) cDNA inserts. Continuous exposure of the pMAMneo/wt‐p53 transfectants to 1 μM, dexamethasone for more than 24 h caused overexpression of wt‐p53 followed by cell death with morphological changes typical of apoptosis. Using the wt‐p53‐inducible HL‐60 cells, we examined the effects of differentiation inducers on the wt‐p53‐dependent apoptosis. All‐trans retinoic acid (all‐trans RA) at 1 nM or granulocyte macrophage colonystimulating factor (GM‐CSF) at 35 pM inhibited the wt‐p53‐induced apoptosis over a 42‐h treatment. The apoptosis inhibition by GM‐CSF, but not alltrans RA, was abolished by specific inhibitors of protein kinase C. These results suggest that extracellular signals involved in the differentiation induction could modulate the wt‐p53‐dependent apoptosis through protein kinase C‐dependent and independent pathways.
Long‐term Exposure to the Anti‐inflammatory Agent Phenylbutazone Induces Kidney Tumors in Rats and Liver Tumors in Mice
Authors: Kari Frank , Bucher John , Haseman Joseph , Eustis Scot , Huff James
Published: August 2005

Long‐term toxicity and carcinogenicity of phenylbutazone, a nonsteroidal anti‐inflammatory drug, were evaluated in F344/N rats and B6C3F1 mice. In 2‐year studies, phenylbutazone was given in corn oil by gavage 5 days per week to groups of 50 rats of each sex at doses of 0, 50, or 100 mg/kg body weight, and to groups of 50 mice at doses of 0, 150, or 300 mg/kg body weight. Body weights and survival were similar among groups. Major target organs are kidneys in rats, and liver in mice. Kidney: inflammation, papillary necrosis, and mineralization in both sexes of rats, and hyperplasia and dilatation of the pelvis epithelium, and cysts in female rats. Uncommon tubular cell tumors of the kidney were found in 13 exposed rats: 5 in the 50 mg group and 4 in the 100 mg group of males; 4 in dosed female rats; none in controls. In female rats, dose‐related increases in hyperplasia of the pelvis transitional epithelium, and 2 carcinomas were discovered. Urinary bladder: papillomas of the transitional epithelium were seen in 2 low‐dose male and in 1 low‐dose female rats. Forestomach: ulcers in rats, with acanthosis, hyperkeratosis, and basal cell hyperplasia in female rats; however, no neoplasms were associated with these lesions. Liver: primarily in male mice exposed to. phenylbutazone, hemorrhage, centrilobular cytomegaly and karyomegaly, fatty metamorphosis, cellular degeneration, and coagulative necrosis were seen; clear cell foci were observed in male mice. In summary, under the conditions of these 2‐year oral intubation studies, phenylbutazone is associated with renal carcinogenicity in rats, as evidenced by increases in tubular cell neoplasms in both sexes. Evidence of carcinogenicity for male mice was shown by increased incidences and multiplicity of liver tumors. No carcinogenic activity was found for female mice.
Comparison of the Chase Effects of Avidin, Streptavidin, Neutravidin, and Avidin‐Ferritin on a Radiolabeled Biotinylated Anti‐tumor Monoclonal Antibody
Authors: Kobayashi Hisataka , Sakahara Harumi , Endo Keigo , Hosono Makoto , Yao Zheng‐Sheng , Toyama Sakuji , Konishi Junji
Published: August 2005

Injection of avidin can decrease the background radioactivity due to a radiolabeled biotinylated monoclonal antibody. We compared the chase effects of avidin, streptavidin, neutravidin, and avidin‐conjugated ferritin on a radiolabeled antitumor monoclonal antibody in tumor‐bearing nude mice. A radioiodine‐labeled biotinylated monoclonal antibody (OST7) was administered to athymic mice bearing osteogenic sarcomas. After 24 h, an avidin, streptavidin, neutravidin or avidin‐conjugated ferritin chaser was intravenously injected into the mice. At 2 h after the chase, the biodistribution of the radiolabeled monoclonal antibody was determined. Clearance from the blood was dose‐dependently accelerated by avidin and its effect was 10‐fold stronger than that of neutravidin or avidinferritin. Streptavidin did not promote clearance of the biotinylated antibody. Avidin was the most effective chasing agent for improving the biodistribution of the radiolabeled biotinylated monoclonal antibody among the four avidin derivatives tested.
Comparison of False Negative Rates among Breast Cancer Screening Modalities with or without Mammography: Miyagi Trial
Authors: Ohuchi Noriaki , Yoshida Koichi , Kimura Michio , Ouchi Akio , Shiiba Ken‐ichi , Ohnuki Koji , Fukao Akira , Abe Rikiya , Matsuno Seiki , Mori Shozo
Published: August 2005

False negative rates were compared in two screening modalities, physical examination with or without mammography, in an intervention study for women aged over 50 in Miyagi Prefecture. Thirty‐five breast cancers were detected in 12,515 subjects who participated in the trial consisting of physical examination and mammography, whereas 44 breast cancers were detected in 50,105 subjects who received physical examination alone, so that the detection rates were 0.28% and 0.09%, respectively. Among 50,061 subjects who received physical examination alone, 8 women were diagnosed as having breast cancer within 12 months after the screening, while only one of 12,480 screenees receiving the combined modality was so diagnosed, implying false negative rates of 15.4% and 2.8%, respectively. When the screening sensitivity in the combined system was analyzed according to each single modality, the false negative rate provided by physical examination with mammography turned out to be 2.8%, significantly lower than that (33.3%) by the physical examination alone. Minimal breast cancers represented 25.7% of all screen‐detected cancers in the combined modality, compared with 9.1% in the modality without mammography. The trial thus indicates that physical examination combined with mammography may be an appropriate modality for breast cancer screening in women aged over 50 on the basis of screening sensitivity.
High‐level Expression of the CD44 Variant Sharing Exon v10 in Renal Cancer
Authors: Kan Masaharu , Aki Masashi , Akiyama Kinya , Naruo Seiichi , Kanayama Hiro‐omi , Kagawa Susumu
Published: August 2005

To examine whether renal cell carcinoma displays altered CD44 expression we performed reverse transcription‐polymerase chain reaction (RT‐PCR) analysis of CD44 in 38 specimens from renal cancer, normal kidney and metastases of 19 patients and 6 renal cancer cell lines. To detect the CD44 variants, we utilized the RT‐PCR Southern blot method. One out of 19 (5.3%) renal cancer specimens expressed a larger molecular weight band than 1 kb by RT‐PCR analysis, in contrast to previous findings in colon and breast cancer. The band patterns in RT‐PCR were different in 14/17 (82.4%) cases between normal kidney and tumors, and a band of about 700 bp was especially marked in 12/17 (70.6%) tumor specimens and 4/6 (66.7%) cell lines. By cloning and sequencing of the 700 bp band, we found that this variant is identical to the CD44 variant sharing only exon v10. Examination by Northern blot analysis has revealed that all tumors express a higher level of CD44 mRNA than paired normal kidneys. These findings suggested that the CD44 variants sharing exon v10 play some role in renal cancer.
A Novel Somatic Point Mutation of the RET Proto‐oncogene in Tumor Tissues of Small Cell Lung Cancer Patients
Authors: Futami Hitoyasu , Egawa Shin‐ichi , Tsukada Toshihiko , Maruyama Kouji , Bandoh Shuji , Noguchi Masayuki , Yamaguchi Ken
Published: August 2005

We examined whether the novel point mutation from GCC (Ala) to GAC (Asp) at codon 664 in exon 11 of RET proto‐oncogene, which we had found in two small cell lung carcinoma (SCLC) cell lines, existed in genomic DNA of tumor tissues of the two SCLC patients from whom these SCLC cell lines were derived. Sequence analysis revealed that point mutation identical to that of the SCLC cell lines was present in amplified alleles of single‐strand conformational variants in genomic DNA of the tumor tissues, whereas it was not detected in genomic DNA of non‐tumor tissues of the patients. These results indicate that this mutation had initially occurred in the SCLC patients and was of somatic origin.
Establishment of a Human Cell Line Secreting Neuron‐specific Enolase from a Primitive Neuroectodermal Tumor of the Retroperitoneal Cavity
Authors: Nakashima Jun , Horiguchi Yutaka , Ueno Munehisa , Nakamura Kaoru , Tachibana Masaaki , Hata Jun‐ichi , Tazaki Hiroshi
Published: August 2005

Primitive neuroectodermal tumor (PNET) is one of the small round cell malignancies of presumed neural crest origin for which an effective treatment has not yet been established. In the present study, a human cell line, designated KU‐9, was established from a 27‐year‐old male patient with PNET of the retroperitoneal cavity and has been successfully maintained in nude mice and in culture. On histological examination, the primary tumor was composed of poorly differentiated small round cells arranged in clusters showing a variety of mitotic changes, and contained Homer‐Wright rosettes. The histopathological appearance of the KU‐9 xenografts was similar to that of the primary tumor. Electron microscopy revealed neurosecretory granules and cytoplasmic processes in the xenograft. No significant amplification of N‐myc gene was observed in the KU‐9 cells. The KU‐9 cells showed chromosome numbers ranging from 56 to 61 with consistent structural abnormalities being add(2)(q31), +add(ll)(pll.2), +add(13)(pll.l), and + del(22)(q12). Cultured KU‐9 cells grew exponentially with a doubling time of about 50 h and a time‐dependent increase in medium levels of neuron‐specific enolase (NSE) was noted. Serum levels of NSE in KU‐9 tumor‐bearing nude mice were significantly elevated and a linear relationship between the serum NSE levels and the tumor NSE content or tumor volume was observed, suggesting that serum levels of NSE may reflect the PNET tumor burden and tumor extent. These results indicate that the KU‐9 cell line provides a reproducible model system which could be useful in gaining some insight into the histogenesis and oncogenesis of PNET and in establishing an effective treatment for PNET.
Expression of Thymidine Phosphorylase in Human Gastric Carcinoma
Authors: Takebayashi Yuji , Miyadera Kazutaka , Akiyama Shin‐ichi , Hokita Shuichi , Yamada Kazutaka , Akiba Suminori , Yamada Yuji , Sumizawa Tomoyuki , Aikou Takashi
Published: August 2005

The activity of thymidine phosphorylase (dThdPase) has heen reported to increase in several types of malignant tumors. Experimental evidence has shown that dThdPase is identical to platelet‐derived endothelial cell growth factor, and that dThdPase has angiogenic activity. We examined the expression of dThdPase to investigate whether the expression of dThdPase correlates with angiogenesis, clinicopathologic features and the prognosis of patients with human gastric carcinomas. Microvessels were assessed by immnnostaining endothelial cells for factor VIII. We counted microvessels in the tumors of 158 patients whose tumors were completely removed surgically. Microvessels were counted in a × 400 field in the most active areas of neovascularization. We purified a monoclonal antibody (TMA‐1) against dThdPase and studied the expression of dThdPase using TMA‐1 in the same serial sections as those used for the detection of factor VIII. The correlation between angiogenesis and dThdPase, and the clinicopathological significance of dThdPase, in patients with gastric carcinoma were examined. The positive expression of dThdPase was more frequent (P<0.001) in gastric carcinomas (67/158, 43.4%) than that in normal tissues (12/158, 7.6%). The average microvessel count in dThdPase‐positive gastric carcinomas was higher (P<0.001) than that in dThdPase‐negative carcinomas. The percentage of gastric carcinoma cells expressing dThdPase was significantly correlated with the microvessel count (P<0.001). Further, the average size of dThdPase‐positive carcinomas was significantly larger (P<0.001) than that of negative carcinomas and the mean microvessel count in dThdPase‐positive gastric carcinomas was also significantly higher (P<0.001) than that in dThdPase‐negative carcinomas. There was a significant correlation between the positive expression of dThdPase and microvessel count (P<0.001) or lymph node metastasis (P=0.013) by multivariate logistic analysis. Further, patients with dThdPase‐positive carcinoma showed a significantly worse prognosis than those with dThdPase‐negative carcinoma overall and in stage III. These findings indicate that the expression of dThdPase in gastric carcinomas is related to progression and metastasis, and this enzyme affects the prognosis of some patients with the disease.
Oncogenic Collaboration of the Cyclin D1 (PRAD1, bcl‐1) Gene with a Mutated p53 and an Activated ras Oncogene in Neoplastic Transformation
Authors: Uchimaru Kaoru , Endo Koichi , Fujinuma Haruko , Zukerberg Lawrence , Arnold Andrew , Motokura Toru
Published: August 2005

Cyclin D1 is one of the key regulators in G1 progression in the cell cycle and is also a candidate oncogene (termed PRAD1 or bcl‐1) in several types of human tumors. We report a collaboration of the cyclin D1 gene with ras and a mutated form of p53 (p53‐mt) in neoplastic transformation. Transfection of cyclin D1 alone or in combination with ras or with p53‐mt was not sufficient for focus formation of rat embryonic fibroblasts. However, focus formation induced by co‐transfection of ras and p53‐mt was enhanced in the presence of the cyclin D1‐expression plasmid. Co‐transfection of ras‐ and p53‐mt‐transformants with the cyclin D1‐expression plasmid resulted in reduced serum dependency in vitro, Furthermore, the transformants expressing exogenous cyclin D1 grew faster than those without the cyclin D1 plasmid when injected into nude mice. These observations strengthen the significance of cyclin D1 overexpression through gene rearrangement or gene amplification observed in human tumors as a step in multistep oncogenesis; deregulated expression of cyclin D1 may reduce the requirement for growth factors and may stimulate in vivo growth.
Effect of MUC1 Mucin, an Anti‐adhesion Molecule, on Tumor Cell Growth
Authors: Makiguchi Yusuke , Hinoda Yuji , Imai Kohzoh
Published: August 2005

MUC1 mucin is expressed in a wide variety of tumors and is considered to function as an anti‐adhesion molecule which inhibits cell‐to‐cell interactions. To reveal the biological significance of this activity in tumor cells, MUC1 cDNA was transfected into EJNIH3T3 cells and human colon cancer cell lines, CHCY1 and DLD1. The in vivo growth rate of MUC1+ (MUC1‐transfected) EJNIH3T3, CHCY1 and DLD1 cells in SCID mice was clearly lower than that of MUC1− (mock transfectant) cells. Several in vitro experiments using MUC1+ EJNIH3T3 cells were performed to analyze the mechanisms for the decreased in vivo tumor growth. It was found that (i) the in vitro growth rate of MUC1+ EJNIH3T3 cells was also decreased compared to that of MUC1− cells, (ii) the DNA synthesis of MUC1+ EJNIH3T3 cells after stimulation with either growth factor (fetal calf serum or bombesin) or extracellular matrix (collagen or fibronectin) was lower than that of MUC1− cells, and (iii) MUC1+ EJNIH3T3 cells grew more slowly than MUC1− cells on both collagen‐ and fibronectin‐coated dishes. These data suggest that MUC1 mucin may regulate tumor cell growth through inhibition of cell‐to‐cell, growth factor‐to‐receptor and cell‐to‐matrix interactions.
Suppressing Effects of 6‐(2,5‐Dichlorophenyl)‐2,4‐diamino‐1,3,5‐triazine and Related Synthetic Compounds on Azoxymethane‐induced Aberrant Crypt Foci in Rat Colon
Authors: Hirose Yoshinobu , Tanaka Takuji , Makita Hiroki , Yang Muzheng , Satoh Kumiko , Hara Akira , Maeda Mitsuaki , Toriyama Hiroyasu Baba , Mori Hideki , Tsuda Hiroyuki
Published: August 2005

The modifying effects of dietary administration of 6‐(2,5‐dichlorophenyl)‐2,4‐diamino‐1,3,5‐triazine and 5 related compounds on the occurrence of azoxymethane (AOM)‐induced colonic aberrant crypt foci (ACF) were investigated in rats. Male F344 rats were given s.c. injections of AOM (15 mg/kg body weight) once a week for 3 weeks to induce ACF. They also received the diet containing 200 ppm test compound for 5 weeks, starting one week before the first dosing of AOM. At the termination of experiment, all of the compounds had caused a significant reduction in ACF frequency, which might be associated with suppression of the expression of proliferation biomarkers. The apoptotic index in the colonic mucosal epithelium of rats killed at 6 h after the first AOM exposure revealed no blocking activity of the compounds.
Expression of Mucin Carbohydrates and Core Proteins in Carcinomas of the Ampulla of Vater: Their Relationship to Prognosis
Authors: Kitamura Hiroshi , Yonezawa Suguru , Tanaka Sadao , Kim Young S. , Sato Eiichi
Published: August 2005

We examined the expression of carbohydrate antigens which are associated with the earliest steps in mucin glycosylation (Tn and sialosyl‐Tn) and the expression of the mucin core protein antigens associated with MUC1 gene product (DF3 antigen) as well as MUC2 gene product (intestinal‐MRP antigen) in tissues from 38 patients with carcinoma of the ampulla of Vater, in order to determine whether these mucin antigens are available as tumor markers or not, and to evaluate whether their expression is correlated with the biological behavior of the carcinomas or not. DF3 antigen showed a relatively high expression rate (61%) in the carcinoma tissues, but was rarely expressed in the non‐neoplastic epithelium around the carcinomas in the region of the ampulla of Vater, Tn and sialosyl‐Tn antigens showed high expression rates in the carcinoma tissues (86% and 84% each), whereas they showed rare or no expression in the non‐neoplastic epithelium around the carcinomas, except for highly restricted expression in the duodenal villous epithelium. The patients with positive DF3 expression in the carcinoma showed significantly poorer survival than those with negative DF3 expression (P<0.05), whereas the patients with positive intestinal‐MRP expression in the carcinoma showed significantly more favorable survival than those with negative intestinal‐MRP expression (P<0.05). The expression rate of DF3 antigen was significantly higher in the cases with deep invasion into the pancreas (89%) than in those with no or minimal invasion (52%) (P<0.05). In contrast, the expression rate of intestinal‐MRP antigen was significantly higher in the cases with no or minimal invasion into the pancreas (38%) than in those with deep invasion (0%) (P<0.05). In conclusion, the expression of DF3, Tn and sialosyl‐Tn antigens is an effective histopathological indicator for carcinomas in the area of the ampulla of Vater, and the expression of DF3 and intestinal‐MRP antigens is a useful indicator of the prognosis of the patients.
Effect of Circulating Antigen on Immunoscintigraphy of Ovarian Cancer Patients Using Anti‐CA125 Monoclonal Antibody
Authors: Sakahara Harumi , Hosono Makoto , Kobayashi Hisataka , Yao Zhengsheng , Saga Tsuneo , Yano Shinsuke , Endo Keigo , Mori Takahide , Konishi Junji
Published: August 2005

The murine monoclonal antibody (mAb) 145–9 recognizes an epitope present on CA125 but different from the epitope defined by the mAb OC125. To evaluate the clinical usefulness of the 145–9 antibody, immunoscintigraphy was performed in ovarian cancer patients and the effect of circulating CA125 on tumor imaging was investigated. Two milligrams (74 MBq) of 111In‐labeled 145–9 was injected intravenously into 11 patients with ovarian cancer. Pre‐injection serum CA125 concentrations were between 166 U/ml and 7414 U/ml. Tumors were visualized in 10 of 11 patients. In two patients, lymph nodes that were not detected by other imaging modalities but were clinically suspected as metastases were visualized. There was no correlation between serum CA125 level and antibody uptake in the tumors. Immune complexes between the antibody and circulating antigen were observed in sera of all the patients, but the fraction of radioactivity in complex form did not correlate well with serum CA125 levels. The immune complexes survived in the circulation and the circulating radiolabel, including immune complexes, was still bound to solid‐phase CA125. The plasma clearance rate and hepatic uptake of the antibody were not significantly affected by circulating CA125. In conclusion, the antibody 145–9 formed complexes with CA125 in vivo but this did not compromise the outcome of antibody imaging. The antibody 145–9 can be used in immunoscintigraphy of ovarian cancer irrespective of serum CA125 level.
Selective Enhancement by Tumor Necrosis Factor‐α of Vascular Permeability of New Blood Vessels Induced with Agarose Hydrogel‐entrapped Meth‐A Fibrosarcoma Cells
Authors: Okada Naoki , Kaneda Yoshihisa , Miyamoto Hajime , Yamamoto Yoko , Mizuguchi Hiroyuki , Tsutsumi Yasuo , Nakagawa Shinsaku , Mayumi Tadanori
Published: August 2005

We have previously developed a simple and quantitative method for assessment of in vivo tumor cell‐induced angiogenesis by means of microencapsulation of tumor cells in agarose hydrogel and mouse hemoglobin ELISA (mHb‐ELISA). In this article, we report that the new blood vessels induced with agarose‐encapsulated tumor cells have the same sensitivity to tumor necrosis factor‐α (TNF‐α) as the original solid‐tumor vessels. Agarose beads (average diameter=200 μm), in which Meth‐A fibrosarcoma cells were microencapsulated, were subcutaneously implanted in non‐syngeneic ddY mice. Ten days later, extensive angiogenesis was observed on the implanted sites of Meth‐A agarose beads, whereas no new blood vessels were induced with cell‐free agarose beads. The vascular permeability of the new blood vessels induced with agarose‐microencapsulated Meth‐A cells was selectively and significantly enhanced by the i.v. injection of TNF‐α, and it reached the maximum level at 2 h after the injection of TNF‐α. At 4 h after the injection of TNF‐α, the vascular permeability was reduced to the basal level. This permeability profile in Meth‐A agarose beads in ddY mice is very similar to that in Meth‐A solid tumor in syngeneic BALB/c mice. On the other hand, TNF‐α‐treatment did not affect the vascular permeability of other normal tissues or inflammatory tissue in ddY mice. These results strongly suggest that the new blood vessels induced with agarose‐microencapsulated tumor cells have the specific characteristics of tumor vessels. Our in vivo angiogenesis assay system should be useful not only to screen anti‐angiogenetic agents, but also to elucidate the mechanism of tumor angiogenesis.
Clonal Heterogeneity in Human Esophageal Squamous Cell Carcinomas on DNA Analysis
Authors: Hori Hikaru , Miyake Satoshi , Akiyama Yoshimitsu , Endo Mitsuo , Yuasa Yasuhito
Published: August 2005

Cancers are thought to arise through multistep accumulation of somatic mutations in the progeny of a single cell. Multiple mutations may induce molecular intratumor heterogeneity. Therefore, we examined molecular clonal heterogeneity in esophageal squamous cell carcinomas. Twenty‐four esophageal squamous cell carcinomas and associated lymph node metastases were examined for microsatellite alterations, and abnormalities of the p53 and transforming growth factor‐β type II receptor (TGF‐β RII) genes. There were eight cases (33%) showing different patterns of loss of heterozygosity in primary tumors and metastatic lymph nodes with microsatellite markers. On the other hand, the abnormalities of p53 were identical in all these cases. No mutation was detected in the simple repeated sequences of the TGF‐β RII gene. These results indicate that molecular clonal heterogeneity exists in esophageal squamous cell carcinomas. Therefore, care is necessary in preoperative genetic diagnosis using biopsy samples.
Metastatic Potential of Lymphoma/Leukemia Cell Lines in SCID Mice Is Closely Related to Expression of CD44
Authors: Kawasaki Nobuhiro , Matsuo Yoshinobu , Yoshino Tadashi , Yanai Hiroyuki , Oka Takashi , Teramoto Norihiro , Liu Cao , Kondo Eisaku , Minowada Jun , Akagi Tadaatsu
Published: August 2005

To investigate whether the lymphocyte homing receptors, adhesion molecules regulating normal lymphocyte traffic, influence the dissemination of lymphoma cells, 24 lymphoma/leukemia cell lines were inoculated into SCID mice subcutaneously, and the correlation between the expression of the adhesion molecules and the metastatic potential of the cell lines was examined. Among the six adhesion molecules examined (LFA‐1, ICAM‐1, CLA, VLA‐4, L‐selectin and CD44), L‐selectin increased the incidence of lymph node metastasis, and CD44 expression was related to both lymph node and organ (hematogenous) metastasis. A monoclonal antibody to the standard form of CD44 (CD44s), Hermes‐3, inhibited the local growth and remote metastasis of CD44+ cell lines. Thus, it is concluded that at least CD44s expression is important in both lymphatic and hematogenous metastasis.
Production of Murine Leukemia RL♂1 Rejection Antigen Peptide pRL1a by Proteolysis of Natural Precursor pRL1b
Authors: Ono Toshiro , Uenaka Akiko , Nakayama Eiichi
Published: August 2005

In this study, we demonstrated that NH2‐terminal Ser and Ile residues of pRL1b (SI‐pRL1a) (SIIPGLPLSL) are not involved in the recognition by RL♂1‐specific cytotoxic T lymphocyte. The sensitization activity observed with pRL1b (SI‐pRL1a) was not greater than that of peptides substituted with irrelevant amino acids at these positions. In serum‐free medium, pRL1a retained sensitization activity, but pRL1b (SI‐pRL1a) did not. Furthermore, addition of bestatin to serumcontaining medium blocked sensitization by pRL1b (SI‐pRL1a). On the other hand, the addition of captopril enhanced it, probably by inhibiting the degradation of pRL1a by ACE. pRL1a‐D peptide with D‐Ile in place of the L‐Ile residue of pRL1a (IPGLPLSL) showed sensitization, but SI‐pRL1a‐2,3D peptide, which has D‐Iles in place of the L‐Ile residues of pRL1b (SI‐pRL1a), and which was not cleaved between the two D‐Iles, did not. The findings suggest that pRL1a is the antigenic peptide bound to Ld molecules and pRL1b (SI‐pRL1a) peptide is its natural precursor, which generates pRL1a via proteolysis.
Expression of Fas Antigen and Its Mediation of Apoptosis in Human Gastric Cancer Cell Lines
Authors: Hayashi Hisae , Tatebe Shigeru , Osaki Mitsuhiko , Goto Akira , Suzuki Yoshimasa , Ito Hisao
Published: August 2005

Fas, a member of the tumor necrosis factor receptor/nerve growth factor receptor family, induces apoptosis by crosslinking with Fas ligand or anti‐Fas antibody in a variety of cultured cells. We examined the expression of Fas antigen and its mediation of apoptosis in six human gastric carcinoma cell lines. Flow cytometric analysis and western blotting revealed relatively high expression of Fas antigen in MKN‐74 (wild‐type p53 gene) and MKN‐45 (wild‐type), followed by MKN‐1 (mutated), MKN‐7 (mutated) and KATO‐III (deleted). MKN‐28 (mutated) showed minimal expression of the antigen. The expression was apparently enhanced by interferon‐γ, except for MKN‐1 and MKN‐28. Anti‐Fas antibody (100 ng/ml) induced nuclear fragmentation characteristic of apoptosis. Apoptosis occurred in a delayed fashion and the apoptotic index at 72 h was approximately 60% in MKN‐74, 35% in MKN‐45, and 20% in MKN‐1 and KATO‐III. A DNA ladder was noted in MKN‐74 at 72 h. Expression levels of P53 and P21Wafl did not change for up to 48 h in MKN‐74. The biological effects did not correlate with endogenous Bcl‐2 expression. These results indicated that a) Fas antigen is variably expressed in human cultured gastric carcinoma cells, b) the protein transduces an apoptotic signal which leads to delayed cell death, and c) susceptibility to the antibody correlates well with the expression level of Fas antigen.
Stromal Expression of MMP‐9 and Urokinase Receptor Is Inversely Associated with Liver Metastasis and with Infiltrating Growth in Human Colorectal Cancer: A Novel Approach from Immune/Inflammatory Aspect
Authors: Takeha Satoshi , Fujiyama Yoshihide , Bamba Tadao , Sorsa Timo , Nagura Hiroshi , Ohtani Haruo
Published: August 2005

MMP‐9 (gelatinase B) and urokinase‐type plasminogen activator receptor (u‐PAR), which are involved in cancer cell invasion and metastasis, are reported to be predominantly expressed by immune/inflammatory cells in human colorectal cancers. To investigate their significance in cancer progression, we morphometrically analyzed the tissue expression of MMP‐9 and u‐PAR among different stages of colorectal cancer. The numbers of MMP‐9‐ and u‐PAR‐positive cells along the invasive margin were significantly smaller in cases with liver metastasis than in cases without liver metastasis, and were also smaller in cases with an infiltrating margin than in cases with an expanding margin. Both variables were larger in colon cancer cases with conspicuous lymphocytic infiltration.These results indicated that the degree of tissue expression of MMP‐9 and u‐PAR by host cells is inversely associated with liver metastasis and an infiltrating growth pattern in human colorectal cancers. Essentially the same results were obtained for the number of macrophages distributed along the invasive margin. We also found that the expression pattern of MMP‐9 was similar to that of MMP‐8 (polymorphonnclear leukocyte collagenase). These data are consistent with clinicopathologic studies of host cells. Therefore, our data suggest a dual role of MMP‐9 and u‐PAR expression in colon cancer tissue; i.e., not only are these proteinases cancer‐promoting factors, hut also they are related to the host defensive mechanism when they are expressed by host cells
Establishment by Adriamycin Exposure of Multidrug‐resistant Rat Ascites Hepatoma AH130 Cells Showing Low DT‐diaphorase Activity and High Cross Resistance to Mitomycins
Authors: Wakusawa Shinya , Nakamura Shigeo , Miyamoto Ken‐ichi
Published: August 2005

A resistant subline (AH130/5A) selected from rat hepatoma AH130 cells after exposure to adriamycin (ADM) showed remarkable resistance to multiple antitumor drugs, including mitomycin C (MMC) and porflromycin (PFM). PFM, vinblastine (VLB), and ADM accumulated in AH130/5A far less than in the parent AH130 (AH130/P) cells. AH130/5A cells showed overexpression of P‐glycoprotein (PGP), an increase in glutathione S‐transferase activity, and a decrease in DT‐diaphorase and glutathione peroxidase activity. The resistance to MMC and VLB of AH130/5A cells was partly reversed by H‐87, an inhibitor of PGP. Buthionine sulfoximine, an inhibitor of glutathione synthase, did not affect the action of MMC. tert‐Butylhydroquinone induced DT‐diaphorase activity, increased PFM uptake, and enhanced the growth‐inhibitory action of MMC in AH130/5A cells. Dicumarol, an inhibitor of DT‐diaphorase, decreased PFM uptake and reduced the growth‐inhibitory action of MMC in AH130/P cells. These results indicated that the adriamycin treatment of hepatoma cells caused multifactorial multidrug resistance involving a decrease in DT‐diaphorase activity.
Experimental Studies on Potentiation of the Antitumor Activity of 5‐Fluorouracil with 3′‐Azido‐3‐deoxythymidine for the Gastric Cancer Cell Line MKN28 in vivo
Authors: Yasuda Chikao , Kato Michio , Kuroda Daisuke , Ohyanagi Harumasa
Published: August 2005

A new method of biochemical modulation of 5‐fluorouracil (5‐FU) with 3‐azido‐3′‐deoxythymidine (AZT)was studied experimentally. Nude mice transplanted with cells of the human gastric cancer cell line MKN28 were divided into 4 groups, i.e., control, 5‐FU, AZT, and 5‐FU plus AZT, and the antitumor activities were compared. Based on the assessment of tumor volume, significant suppression of tumor growth was observed in the 5‐FU and 5‐FU plus AZT groups (P<0.05, P<0.01, versus control, respectively). The thymidylate synthase (TS) inhibition rate, an index of inhibition of the de novo pathway, was significantly higher in the 5‐FU and 5‐FU plus AZT groups than in the control group (P<0.01), but it did not differ from the control in the AZT group. TS‐bound FdUMP tended to be higher in the 5‐FU pins AZT group than in the 5‐FU group. The activity of thymidine kinase (TK) and the uptake ratio of 5‐hromo‐2′‐deoxyuridine (BrdU), indices of salvage pathway activity, were significantly lower in the AZT and 5‐FU plus AZT groups than in the control group (TK, P<0.05, P< 0.01; uptake ratio of BrdU, P< 0.01, P< 0.05, respectively). There were slight losses of body weight in the 5‐FU and 5‐FU plus AZT groups compared with that in the control group, but no difference between the AZT and control groups in weight loss. These findings suggest that addition of AZT plays an important role in potentiating the antitumor activity of 5‐FU through both blockage of a compensatory increase of activity in the salvage pathway and also an increase in TS‐bound FdUMP, anid has no adverse effects. Thus, the combination of 5‐FU and AZT could be useful as a new modality in gastric cancer chemotherapy.
Significant Correlation of Telomerase Activity in Thyroid Papillary Carcinomas with Cell Differentiation, Proliferation and Extrathyroidal Extension
Authors: Okayasu Isao , Osakabe Toji , Fujiwara Mutsunori , Fukuda Hiroki , Kato Motonobu , Oshimura Mitsuo
Published: August 2005

Telomerase activity was examined by telomeric repeat amplification protocol assay in thyroid disease states, including adenomas and carcinomas, and correlated with clinicopathological features. Of a total of 26 papillary carcinomas, 16 cases (61.5%) were positive, with the poorly differentiated subtype being predominant (P<0.05). A significantly more shortened terminal restriction fragment length (P<0.05), higher incidence of extrathyroidal extension (P< 0.001), and more elevated Ki‐67 labeling indices (P<0.002) were also found in telomerase‐positive than in telomerase‐negative papillary carcinomas. Of four follicular carcinomas, 3 cases (75.0%) were positive. Positive telomerase activity in follicular adenomas (9/23 cases, 39.1%) and lymphocytic thyroiditis (12/22 cases, 54.5%) appeared to be mainly caused by infiltrating lymphocytes. However, three cases of atypical adenoma with relatively increased Ki‐67 labeling indices were positive, suggesting a possibility of malignant potential. The good correlations with extrathyroidal invasiveness, Ki‐67 labeling indices and poor differentiation of papillary carcinomas, established by multivariatc analysis, suggest that this parameter might have potential application in the estimation of tumor progession and prognosis, and in clinical management
MIB1‐determined Proliferative Activity in Intraductal Components and Prognosis of Invasive Ductal Breast Carcinoma
Authors: Imamura Hiroshi , Haga Shunsuke , Shimizu Tadao , Watanabe Osamu , Kajiwara Tetsuro , Aiba Motohiko
Published: August 2005

Intraductal components of breast carcinoma may have prognostic significance. In this study, we divided 181 invasive ductal breast carcinomas into comedo and non‐comedo groups based on intraductal component morphology, and differences between the two groups in clinicopathological variables, including proliferative activity and survival, were assessed. Proliferative activity was evaluated by using MIB1 antibody, which reacts with the cell‐proliferation‐associated Ki‐67 antigen, and was expressed as the number of MIBl‐positive nuclei per 1000 cancer cells in intraductal components (MIB1 labeling index). We also investigated which variables had an impact on survival. The comedo group showed a significantly higher MIB1 labeling index than the non‐comedo group (P < 0.0001). The differences in disease‐free and overall survival between the two groups were not significant (P=0.2477, P=0.2069). Multivariate analysis of the entire series showed the MIB1 labeling index to be an independent prognostic factor predicting both disease‐free survival and overall survival (P=0.0160, P=0.0035). When multivariate analysis was repeated separately for the non‐comedo and comedo groups, the MIB1 labeling index remained the most important variable predicting disease‐free and overall survival in the non‐comedo group (P=0.0122, P=0.0040). Moreover, non‐comedo patients with a high MIB1 labeling index had significantly shorter disease‐free and overall survival than those with a low MIB1 labeling index (P=0.0040, P=0.0402). These findings imply that MIB1‐determined proliferative activity of intraductal components is an independent predictor of survival, and is the most important predictor in non‐comedo cases.