bluuebunny/crossref_metadata_2025 huggingface dataset:
Monoclonal Antibody against Rat Renal Cell Tumor‐associated Antigen as a New Tool for the Analysis of Renal Tumorigenesis
Authors: Konishi Noboru , Kitahori Yoshiteru , Shimoyama Taketo , Lin Jung‐Chung , Hiasa Yoshio
Published: August 2005

A monoclonal antibody, NR‐2 MAb, against one of the rat renal cell tumor‐associated antigens was developed. NR‐2 MAb belonged to IgM class and recognized a polypeptide of 81,000 daltons designated as NR‐2 antigen, which is of non‐glycoprotein nature with a pI of 4.6. NR‐2 MAb was employed to probe the histogenesis of renal cell tumors in rats treated with N‐ethyl‐N‐hydroxyethylnitrosamine followed by trisodium nitrilotriacetate monohydrate. Immunohistochemical analysis indicated that NR‐2 antigen was expressed in simple hyperplasia, adenomatous hyperplasia and renal cell tumors. Both clear cells and basophilic cells of the simple hyperplasia showed equally strong positive reactions with NR‐2 MAb, whereas the vacnolated epithelium was negative. Furthermore, the proximal tubules in nontumorous areas also expressed NR‐2 antigen, suggesting that the hyperplastic lesions which eventually lead to renal cell tumors may derive from cpithelia of proximal tubules and not directly from vacuolated epithelium. Such NR‐2 antigen‐positive epithelia of proximal tubules seem to be initiated cells. NR‐2 MAb also cross‐reacted with preneoplastic liver lesions.
Serial Measurement of Sister Chromatid Exchanges in the Peripheral Lymphocytes of Patients with Lung Cancer Receiving Chemotherapy in Relation to Bone Marrow Toxicity
Authors: Shinkai Tetsu , Saijo Nagahiro , Eguchi Kenji , Sasaki Yasutsuna , Tamura Tomohide , Fujiwara Yasuhiro , Kojima Akira , Nakagawa Kazuhiko , Minato Kouichi , Nakajima Takashi , Suemasu Keiichi
Published: August 2005

The frequencies of sister chromatid exchanges (SCE) in the peripheral lymphocytes of patients with lung cancer before and after initial chemotherapy were serially measured and the correlation between SCE frequencies and bone marrow toxicity was evaluated. The addition of mitomycin C to vindesine plus cisplatin increased SCE frequencies significantly in patients with non‐small cell lung cancer. A significant increase in frequencies of SCE was observed in patients treated with cyclophosphamide plus adriamycin plus vincristine as compared with those treated with cisplatin plus etoposide in small cell lung cancer. A significant inverse correlation was observed between SCE frequencies in the peripheral lymphocytes 7 days after treatment (x) and the nadir value/pretreatment value of platelets (y)(r= ‐0.685, P= 0.0007, y= 0.842–0.022x). The relation between SCE frequency and leukopenia showed the same trend as thrombocytopenia, but the correlation was not statistically significant (r= ‐ 0.444, P= 0.057). SCE assay may have potential clinical use for the prediction of chemotherapyinduced thrombocytopenia.
Enhancement of Both Intracellular Uptake and Antitumor Action of Cisplatinum on Human Neuroblastoma Cells by Encapsulation in Liposomes
Authors: Kamio Yoshiro , Kato Hiroshi , Kishikawa Teruaki , Toda Takashi , Sasaki Shingi , Ito Jin‐ichi , Kato Taiji , Tanaka Ryo
Published: August 2005

Phospholipid vesicles (phosphatidylcholine: phosphatidylserine: cholesterol=6:2:3 in molar ratio) with a small unilamellar structure were used as drug carriers for introducing cis‐diamminedichloroplatinum (CDDP) into human neuroblastoma cells, IMR‐32, GOTO, Nagai, and TGW. DNA synthesis of IMR‐32 cells among the human neuroblastoma cell lines was inhibited most strongly by CDDP‐liposomes. CDDP‐liposomes dose‐dependently inhibited the DNA synthesis of IMR‐32 in a similar fashion to that observed with free CDDP, but the drug concentration required to induce 50% inhibition of DNA synthesis for CDDP‐liposomes (IC50: 0.7 μg CDDP/ml) was 1/3 of the IC50 for free CDDP (2.0 μg CDDP/ml). In support of the marked growth‐inhibitory action of CDDP‐liposomes, the intracellular incorporation rate of CDDP‐liposomes was 3‐fold higher when liposomes were used as carriers than when free CDDP was directly applied. CDDP‐liposomes showed a stronger growth inhibition on IMR‐32 cells at a high cell density than at a low density in culture. CDDP‐liposomes were rapidly incorporated by IMR‐32 cells within 5 min, resulting in the inhibition of DNA synthesis to 40% of the control. Swiss albino mouse 3T3 cells were less inhibited by CDDP‐liposomes than by free CDDP, suggesting that encapsulation of CDDP in liposomes decreases cytotoxicity to normal cells.
Coexpression of Platelet‐derived Growth Factor (PDGF) A‐Chain and PDGF Receptor Genes in Human Gastric Carcinomas
Authors: Tsuda Toshitaka , Yoshida Kazuhiro , Tsujino Tetsuhiro , Nakayama Hirofumi , Kajiyama Goro , Tahara Eiichi
Published: August 2005

In this study we examined the expression of platelet‐derived growth factor (PDGF) A‐chain and PDGF receptor genes in seven human gastric carcinoma cell lines and 15 gastric carcinoma tissues. Expression of mRNA for PDGF A‐chain was found in all gastric cell lines and all gastric carcinoma tissues. Two of the seven gastric carcinoma cell lines expressed PDGF receptor mRNA. Out of the 15 gastric carcinoma tissues, eight showed enhanced expression of PDGF receptor mRNA and all of them demonstrated prominent fibrous stroma. Moreover, the incidence of enhanced expression of PDGF receptor mRNA was higher in scirrhous carcinoma than in well differentiated adenocarcinoma. These results strongly suggest that PDGF produced by tumor cells acts as a paracrine growth factor for production of fibrous stroma in gastric carcinomas
Risk Factors for Leukemia Occurrence among Growth Hormone Users
Authors: Watanabe Shaw , Yamaguchi Naohito , Tsunematsu Yukiko , Komiyama Atsushi
Published: August 2005

A matched case‐control study on leukemia occurrence among growth hormone (GH) users was performed to elucidate any risk factor for leukemia. The total doses of GH and administration of other hormones, such as thyroxine, gonadotropin, gonadal hormones, glucocorticoid and anabolic steroid, did not lead to any apparent difference between cases studied and controls. Neither doses of diagnostic X‐rays nor therapeutic cranial irradiation was related to leukemia risk. Scintigraphy for thyroid function using radioactive iodide was the only significant risk factor (P=0.03). Hematological changes and liver function before and one month after GH administration were compared and the cases studied showed more proliferative reaction of white blood cells with rapid increase of neutrophils, but lymphocyte response was variable. Lactic acid dehydrogenase, alkaline phosphatase, glutamic oxalacetic transaminase, glutamic pyruvic transaminase tended to decrease after GH treatment when the patients had shown abnormally increased levels. Other environmental and familial factors did not show any abnormal clusters.
Hematologic and Cytogenetic Findings in Myelodysplastic Syndromes Treated with Recombinant Human Granulocyte Colony‐stimulating Factor
Authors: Ohyashiki Kazuma , Ohyashiki Junko H. , Toyama Keisuke , Takaku Fumimaro
Published: August 2005

We administered recombinant human granulocyte colony‐stimulating factor (rhG‐CSF) to four patients with myelodysplastic syndrome (MDS) and three patients with non‐MDS (two malignant lymphoma and one lung cancer) as a part of a phase II trial and analyzed the effects of rhG‐CSF on the neoplastic cells of MDS by performing sequential chromosome analyses on the bone marrow cells. A greater than 3‐fold increase in neutrophil count was observed in the MDS patients after rhG‐CSF infusions, whereas the number of blasts in the bone marrow did not increase and none of them progressed into the leukemic phase. After rhG‐CSF treatment, the bone marrow cells obtained from patients without MDS did not show any particular chromosome abnormalities such as chromosomal breakage. On the contrary, two of the four MDS patients with acquired chromosome abnormalities showed a change in the frequency of marrow cells with clonal abnormalities after rhG‐CSF treatment; the proportion of metaphase cells with additional numerical chromosome abnormalities decreased in these two MDS patients. After discontinuation of the treatment, the constitution of marrow cells with chromosome changes reverted to that before treatment. The remaining two MDS patients did not show any particular chromosome changes after the rhG‐CSF treatment, indicating that rhG‐CSF may not promote the characteristics of dyshematopoiesis in MDS, and act on cells derived from an MDS clone

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The Tumor Rejection Antigen Separated from Rous Sarcoma Virus‐induced Murine Fibrosarcoma Exhibits a Molecular Weight of Approximately 60 kD but Differs from Functional pp60src
Authors: Suda Takashi , Shimizu Jun , Muramatsu Minoru , Kimura Kazuhiro , Yoshida Takato O. , Fukami Yasuo , Fujiwara Hiromi , Hamaoka Toshiyuki
Published: August 2005

The tumor antigen capable of inducing tumor resistance (tumor rejection antigen; TRA) was obtained in a solubilized form by sodium dodecyl sulfate (SDS) extraction of plasma membrane fraction from Rous sarcoma virus (RSV)‐induced CSA1M fibrosarcoma cells (BALB/c origin). Analyses by Sephacryl S‐300 gel filtration and SDS‐polyacrylamide gel electrophoresis revealed that TRA activity was recovered in the fraction with a molecular weight of approximately 60 kD. Unfractionated crude SDS‐solubilized preparation contained gp70 as detected by rabbit anti‐gp70 antiserum, whereas such reactivity was lost in the fraction exhibiting the molecular weight of about 60 kD. Since this fraction retained pp60src activity, the relation of TRA to pp60src was further investigated. pp60v‐src was also obtained from the lysate of v‐src‐expressing yeast transformant. Immunization of BALB/c mice with such pp60v‐src‐containing lysate failed to induce any significant tumor protection. The above 60 kD fraction of CSA1M solubilized antigens was allowed to bind to Sepharose beads coupled with anti‐pp60src monoclonal antibody and separated into the bead‐bound and bead‐unbound fractions. The bead‐bound fraction that was recovered from pp60src‐binding beads (pp60src‐positive fraction) did not exhibit the TRA activity. In contrast, immunization with the fraction depleted of pp60src activity (bead‐unbound fraction) resulted in potent tumor protection. These results indicate that the solubilized membranous component(s) of CSA1M with a molecular weight of approximately 60 kD, which is distinct from functional pp60src, functions as the TRA against RSV‐induced CSA1M tumor cells.
Enhancement of Cytotoxicity of Cisplatin in vitro by Recombinant Human Tumor Necrosis Factor and/or Recombinant Human Interferon‐α, β and ‐γ
Authors: Kim Chang‐Min , Hong Weon‐Seon , Lee Jhin‐Oh , Rang Tae‐Woong , Kim Young‐Whan , Song Jae‐Kwan , Yun Taik‐Koo , Kim Chung‐Yong
Published: August 2005

This study was conducted to investigate the modulatory effects of recombinant human tumor necrosis factor (rH‐TNF) and recombinant human interferon (rH‐IFN)‐α, ‐β and ‐γ, either alone or in combination, on the cytotoxicity of cisplatin, using MTT assay, against MKN‐45 (human stomach adenocarcinoma). MKN‐45 was resistant to rH‐TNF even at doses up to 103 U/ml. rH‐IFN‐γ inhibited the survival of MKN‐45 dose‐dependently, while rH‐IFN‐α and ‐β did not inhibit the survival of MKN‐45 even at the highest concentrations tested (104 U/ml). Combination of rH‐TNF with rH‐IFN‐α, ‐β or ‐γ did not significantly inhibit the survival of MKN‐45, except for a combination of 10 U/ral of rH‐TNF and 103 U/ml of rH‐IFN‐γ (P<0.05). Cisplatin inhibited the survival of MKN‐45 dose‐dependently. By the simultaneous combination of cisplatin with rH‐TNF and/or rH‐IFN‐α, ‐β or γ, cytotoxicity of cisplatin was enhanced and the combination effects were additive. The effects of rH‐TNF and rH‐IFN‐α, β and ‐γ on the modification of cytotoxicity of cisplatin were evaluated in terms of modification index (MI), demonstrating that rH‐TNF, rH‐IFN‐α, ‐β and ‐γ all augmented the cytotoxicity of cisplatin: MI values at 103 U/ml of rH‐IFN‐α, ‐β and ‐γ were 1.4, 1.4 and 2.3, respectively; those at the same concentrations of rH‐IFN‐α, ‐β and ‐γ in the presence of 10 U/ml of rH‐TNF were 3.6, 2.5 and 5.1, respectively. These results demonstrating that the cytotoxicity of cisplatin was enhanced by rH‐TNF and/or rH‐IFN‐α, ‐β or ‐γ suggest that cancer may be more effectively treated with the combination of cisplatin with these biological response modifiers than with cisplatin alone.
Amplification of Both c‐myc and c‐raf‐1 Oncogenes in a Human Osteosarcoma
Authors: Ikeda Shogo , Sumii Hiroshi , Akiyama Kosuke , Watanabe Sekiko , Ito Shiro , Inoue Hajime , Takechi Hideo , Tanabe Gozo , Oda Takuzo
Published: August 2005

Fourteen human bone and soft part tumor tissues were screened by Southern blot hybridization using five oncogene probes (c‐myc, c‐K‐ras, c‐fos, c‐raf‐l, and N‐myc). Amplification of c‐myc was found in two osteosarcomas and one malignant fibrous histiocytoma. One of these osteosarcomas had amplified c‐raf‐1 gene. Rearrangement of the amplified gene was not observed. This is the first report of c‐raf‐1 amplification in human cancer tissues.
Insulin and Insulin‐like Growth Factor 1 Stimulate Proliferation of Metastatic Variants of Colon Carcinoma 26
Authors: Koenuma Mitsuo , Yamori Takao , Tsuruo Takashi
Published: August 2005

The proliferation rate of malignant cells in vivo is one of the important factors which affect the formation of tumor metastasis. A highly metastatic variant of mouse colon adenocarcinoma 26 (NL‐ 17) grew more rapidly than a low‐metastatic variant (NL‐44) both in vitro and in vivo. The effect of growth factors on the proliferation of NL‐17 and NL‐44 cells was examined in serum‐free medium. Among growth factors examined, human insulin and insulin‐like growth factor 1 (IGF‐1), which were produced by gene engineering techniques, stimulated the growth of metastatic NL‐17 and NL‐44 cells as determined by thymidine incorporation and cell counts. DNA synthesis and cell proliferation of the high‐metastatic NL‐17 was stimulated to a greater extent by insulin and IGF‐1 than those of the low‐metastatic NL‐44. These findings suggest that circulating growth factors could enhance the formation of tumor metastasis. Scatchard analysis of [125I]IGF‐1 binding to NL‐17 and NL‐44 showed that each cell line had an almost equal number of IGF‐1 receptors (1.37 × 105/cell and 1.26 × 105/cell, respectively), which had similar dissociation constants (8.94×10−10M and 9.54×10−10M, respectively). Since the number and affinity of IGF‐1 receptors are equivalent between low‐ and high‐metastatic cells, the intracellular events which result in the cell growth after binding of IGF‐1 may differ between NL‐17 and NL‐44 cells.
Toxicological Aspects of a Novel 9‐Aminoanthracycline, SM‐5887
Authors: Morisada Shinya , Yanagi Yoshikazu , Kashiwazaki Yasuo , Fukui Masaru
Published: August 2005

The toxicological characteristics of SM‐5887 were evaluated in mice after a bolus intravenous injection, and compared with those of adriamycin (ADR). The acute toxic signs observed after SM‐5887 administration were body weight decrease, ataxia, hair loss, and myelosuppression. They were qualitatively comparable to those induced by ADR. The 50% lethal dose values determined by 14‐day observation after drug administration were in the range of 32 to 50 mg/kg for SM‐5887 and 16 to more than 20 mg/kg for ADR in four strains of mice. The maximum tolerated doses (MTD) were estimated to be 25 mg/kg for SM‐5887 and 12.5 mg/kg for ADR (no death or body weight loss of more than 3 g occurred). When 14‐day survivors were further observed until 90 days after drug administration, ADR frequently and dose‐independently showed delayed‐type lethal toxicity at doses of more than 10 mg/kg, whereas SM‐5887 did not. The myelosuppression of SM‐5887 was more severe even at a half of the MTD than that of ADR at the MTD, but its recovery was more rapid than that after ADR, In addition, when the drugs were injected into the subplantar region of mouse hind paws, ADR induced a severe inflammatory reaction, whereas SM‐5887 yielded only a slight one. The data suggest that toxic effects of SM‐5887 are more reversible and more controllable than those of ADR.
Clonal T‐cell Lymphoproliferation Containing Epstein‐Barr (EB) Virus DNA in a Patient with Chronic Active EB Virus Infection
Authors: Ishihara Shigehiko , Tawa Akio , Yumura‐Yagi Keiko , Murata Mitsunori , Hara Jun‐ichi , Yabuuchi Hyakuji , Hirai Kanji , Kawa‐Ha Keisei
Published: August 2005

A 10‐year‐old boy with chronic active Epstein‐Barr virus (EBV) infection developed T‐cell lymphoproliferation in the terminal stage of hepatic failure. The phenotypes of the proliferating lymphocytes were CD3±, CD4± and HLA‐DR±. The genomic DNAs from these cells demonstrated two rearranged T‐cell receptor μ‐chain genes and contained the EBV genome. These findings indicate that EBV can infect T lymphocytes and might cause clonal T‐cell lymphoproliferation.
Mutations of N‐ras Oncogene in Myelodysplastic Syndromes and Lcukemias Detected by Polymerase Chain Reaction
Authors: Mano Hiroyuki , Ishikawa Fuyuki , Hirai Hisamaru , Takaku Fumimaro
Published: August 2005

By using polymerase chain reaction and differential hybridization, point mutations of N‐ras oncogene were investigated among 43 patients with hematopoietic malignancies. Six cases were revealed to carry mutational N‐ras oncogenes. One case with acute myelocytic leukemia was found to carry a mutation at codon 13, although this case was in complete remission.
The Significance of Low Serum Pepsinogen Levels to Detect Stomach Cancer Associated with Extensive Chronic Gastritis in Japanese Subjects
Authors: Miki Kazumasa , Ichinose Masao , Kawamura Norio , Matsushima Masashi , Ahmad Haron Bin , Kimura Masayoshi , Sano Junjiro , Tashiro Takao , Kakei Nobuyuki , Oka Hiroshi , Furihata Chie , Takahashi Kenji
Published: August 2005

Serum pepsinogen levels were measured in 137 stomach cancer patients and compared with those of 288 normal cancer‐free subjects. The serum pepsinogen levels of stomach cancer patients, especially pepsinogen I and the pepsinogen I/pepsinogen II ratio were significantly lower than those of normal controls and correlated well with the extent of chronic gastritis associated with the cancerous stomach. These results were in good accordance with the results of previous studies indicating that the cancer derived from the stomach where chronic gastritis/intestinal metaplasia is extensive. The high sensitivity and specificity of this non‐invasive serum test to detect chronic gastritis suggested the possibility of its application to the mass screening of stomach cancer.
Combined Effects of Differentiation‐inducing Factor and Other Cytokines on Induction of Differentiation of Mouse Myeloid Leukemic Cells
Authors: Yamamoto‐Yamaguchi Yuri , Tomida Mikio , Hozumi Motoo , Maurer Hans Rainer , Okabe Tetsuro , Takaku Fumimaro
Published: August 2005

Mouse myeloid leukemic M1 cells are induced to differentiate into macrophage‐like cells by differentiation‐inducing factors (D‐factors) and granulocyte colony‐stimulating factor. We examined the effects of recombinant human tumor necrosis factor (rTNF), lymphotoxin (rLT) and interleukin 1 (rIL‐1) on the induction of differentiation of M1 cells, compared with the effects of D‐factor purified from the conditioned medium of mouse Ehrlich as cites tumor cells and recombinant human granulocyte colony‐stimulating factor (rG‐CSF). rIL‐1 induced phagocytic activity, a typical marker of cell differentiation, in at most 30% of M1 cells at concentrations ranging from 10‐10M to 10‐7M. The differentiation‐inducing activity of rIL‐1 was similar to that of rG‐CSF and less than that of D‐factor. rTNF induced phagocytic activity in 14% of M1 cells only at a high concentration (10‐7M). rLT did not induce differentiation of the cells even at 10‐7M. rTNF stimulated induction of differentiation of M1 cells by D‐factor, rG‐CSF or rIL‐1 by two or three fold. The combination of any two of the cytokines D‐factor, rG‐CSF and rIL‐1 induced differentiation of M1 cells more efficiently than any of these cytokines alone. Moreover, the combination of three cytokines rG‐CSF, rIL‐1 and rTNF, all of which are known to be produced by macrophages, was more effective than the combination of any two of these cytokines in induction of differentiation of M1 cells.
Changes in the Quantity and Activity of Cytochrome P‐450 Isozymes in Primary Cultured Rat Hepatocytes
Authors: Namiki Masayuki , Degawa Masakuni , Masuko Takashi , Hashimoto Yoshiyuki
Published: August 2005

Hepatocytes from male Spragne‐Dawley rats pretreated with a cytochrome P‐450 inducer, 3‐methoxy‐4‐aminoazobenzene (3‐MeO‐AAB), 3‐methylcholanthrene (MC) or phenobarbital (PB), were cultured in vitro, and changes in the quantity and activity of microsomal cytochrome P‐450 isozymes in the cells were determined by means of immunochemical methods and a bacterial mutation test, respectively. The results of enzyme‐linked immunosorbent assay using monoclonal antibodies against rat P‐450 isozymes ‐ test using Salmonella typhimurium TA 98 and carcinogenic aromatic amines. These results indicate that microsomal cytochrome P‐450c in primary cultured rat hepatocytes is more stable in culture, in terms of both quantity and activity, than cytochrome P‐450d and P‐450b/e.
Tumor‐specific T Cells which Form Clusters with Dendritic Cells and Tumor Cells and Deliver Macrophage‐activating Factors
Authors: Yamaguchi Yasunori , Inaba Kayo , Kawai Jun , Kato Takuma , Nakamura Shinji , Uno Kazuko , Muramatsu Shigeru
Published: August 2005

T cells prepared from tumor (Meth A)‐bearing mice were cocultured with homologous tumor cells and splenic dendritic cells to enrich tumor‐specific T cells by the separation of clusters. T blasts generated from clusters were capable of inhibiting the in vivo tumor cell growth. The culture supernatant of clustering cells (CLSN) was effective in activating macrophages (MØ) to be cytostatic and cytocidal against tumor cells. Moreover, it was found that CLSN contains at least 3 distinct factors; one was identified as interferon‐γ (IFN‐γ), and the others are so far unidentified, but one acts synergistically with IFN‐γ, possibly as the second signal, and the other cooperates with lipopolysaccharide but not with IFN‐γ. We propose that the tumor‐specific T cells secrete soluble mediators which cooperate with each other in MØ activation against tumor cells.
Enhanced Antitumor Effect of 5′‐Deoxy‐5‐fluorouridine by Oral Administration with l‐Cysteine
Authors: Iigo Masaaki , Nakajima Yoko , Araki Eiji , Hoshi Akio
Published: August 2005

When given orally in combination with l‐cysteine, 5′‐deoxy‐5‐fluorouridine (DFUR) brought about a significant reduction in the growth of adenocarcinoma 755 and a significant prolongation of life‐span in mice bearing Lewis lung carcinoma without increased toxicity to the host as compared with DFUR alone, though l‐cysteine alone did not show an appreciable antitnmor activity. Moreover, the combination of DFUR and l‐cysteine resulted in a marked retardation of growth of human colon tumor LS174T transplanted into nude mice. Thus, the potency of DFUR was increased by l‐cysteine. Pharmacokinetic studies revealed that after DFUR administration, plasma DFUR and 5‐fluorouracil (5‐FU) levels rapidly declined, but that, in the combination with l‐cysteine, the plasma clearances of DFUR and 5‐FU were slowed down considerably. In the tumor, DFUR and 5‐FU levels were similar to those in the plasma. Such a prolongation of DFUR and 5‐FU levels in plasma and tumor may produce the enhancement of antitumor effect seen with the combination of DFUR and l‐cysteine.
Inhibitory Action of Transforming Growth Factor‐β on Induction of Differentiation of Myeloid Leukemia Cells
Authors: Okabe‐Kado Junko , Honma Yoshio , Hayashi Moriaki , Hozumi Motoo
Published: August 2005

The effect of transforming growth factor‐β (TGF‐β) on induction of differentiation of mouse myeloid leukemia M1 cells was investigated. TGF‐β1 induced adherence of M1 cells to plastic dishes and inhibited their proliferation. However, it did not induce differentiation‐associated properties, such as phagocytic activity, lysozyme activity or morphological maturation. TGF‐β1 also caused dosedependent inhibition of dexamethasone‐induced differentiation of M1 cells. The inhibitory activity of TGF‐β1 was 20 times that of TGF‐β2 on M1 cells. These results suggest that TGF‐β1 inhibits proliferation and dexamethasone‐induced differentiation of M1 cells by interacting with receptors that can distinguish between TGF‐β1 and TGF‐β2. TGF‐β1 had a much lower inhibitory effect on the growth of a variant M1 cell clone, which was resistant to differentiation inducers, and it did not induce adherence of the resistant M1 cells.
Influence of Tamoxifen‐Medroxyprogesterone Sequential Therapy on Estrogen and Progesterone Receptor Contents of Breast Cancer
Authors: Noguchi Shinzaburo , Yamamoto Hitoshi , Inaji Hideo , Imaoka Shingi , Koyama Hiroki
Published: August 2005

The influence of tamoxifen (TAM) and medroxyprogesterone acetate (MPA) sequential administration on the estrogen receptor (ER) and progesterone receptor (PR) contents of breast cancer was studied in 68 patients with operable breast cancer. TAM was used as a primer of PR induction in order to enhance the effects of MPA. Half of the patients (n=34) were preoperatively treated with TAM (20 nig/day for 7 days) and sequentially with MPA (1200 mg/day for 17 (median) days). ER and PR of surgical specimens were assayed by enzyme immunoassay and the results were compared with those obtained from the other half of the patients (n=34), who had not received any treatment before surgery. TAM‐MPA treatment significantly lowered PR in the cytosol regardless of the menopausal status. On the other hand, TAM‐MPA treatment significantly lowered ER in the cytosol only in the postmenopausals but not in the premenopansals. These results demonstrate that reduction of ER provoked by TAM‐MPA treatment is dependent on menopausal status.
A Correlation between the Expression of CD 8 Antigen and Specific Cytotoxicity of Tumor‐infiltrating Lymphocytes
Authors: Okada Yoshiaki , Yahata Gohki , Takeuchi Shoshichi , Seidoh Tsutomu , Tanaka Kenichi
Published: August 2005

Tumor‐infiltrating lymphocytes (TIL) from six gynecologic malignant tumors (two uterine cervical cancers, two ovarian serous cystadenocarcinomas, and two uterine corpus cancers), cultured in the presence of recombinant interleukin 2, were assayed for their cytotoxic activities against various fresh tumor cells including autologous tumors. A clear correlation between phenotype and cytotoxic activity of TIL was observed. Four of six TIL preparations exhibited strong cytotoxic activity against autologous fresh tumor target cells, and were all CD8±. In contrast, cytotoxic activity was not detected in any of the CD4± TIL preparations. The cytotoxic activities of the CD8± TIL preparations were highly specific; only autologous fresh tumor cells were lysed. This result is consistent with the notion that TIL are of a different cell lineage from lympholdne‐activated killer cells which are antigen‐nonspecific and CD8‐. Instead, TIL appear to be of cytotoxic T cell lineage that is highly antigen‐specific and CD8±. To explore the potential for clinical use, we have attempted to augment the cytotoxic activities of these CD8± TIL by treatment of the target tumor cells with gamma interferon (IFN) in vitro, hoping that elevated expression of MHC class I gene products on the cell surface would enhance their recognition. It was observed that brief treatment of freshly prepared tumor cells in vitro with gamma‐IFN resulted in augmentation of the expression of MHC class I gene products, and the treated tumor cells were more susceptible to lysis by TIL than untreated cells.
Inhibition of Colony Formation of Drug‐resistant Human Tumor Cell Lines by Combinations of Interleukin‐2‐activated Killer Cells and Antitumor Drugs
Authors: Ohtsu Atsushi , Sasaki Yasutsuna , Tamura Tomohide , Fujiwara Yasuhiro , Ohe Yuichiro , Minato Koichi , Nakagawa Kazuhiko , Bungo Masami , Saijo Nagahiro
Published: August 2005

The cytotoxicity of interleukin‐2‐activated killer (LAK) cells with or without anticancer drugs against cell lines with acquired drug resistance was evaluated in vitro by colony assay. Human non‐small cell lung cancer cell lines, PC‐9 and PC‐14, human leukemia cell line, K‐562, and their sublines resistant to cisplatin (CDDP), PC‐9/CDDP and PC‐14/CDDP, and to adriamycin (ADM), K‐562/ADM, were used as target cells. PC‐9/CDDP demonstrated a marked increase in susceptibility to killing by both peripheral blood lymphocytes (PBL) and LAK cells, as compared to the parental cell line, PC‐9. The cytotoxicity of PBL and LAK cells against PC‐14/CDDP and K‐562/ADM was similar to that against their parental cell lines. Moreover, the combination of LAK and CDDP had a synergistic effect on PC‐14 and PC‐14/CDDP.
The Effects of Vincristine and Doxorubicin on the Clonogenic Cells of a Human Lung Cancer Cell Line in Methylcellulose and Suspension Culture
Authors: Yamashita Yoko , Nara Nobuo , Aoki Nobuo
Published: August 2005

The effects of vincristine (VCR) and doxorubicin (DOX) on the growth of an established line of human lung cancer cells, PC9, were studied in methylcellulose and suspension cultures. The secondary colony formation in methylcellulose and recovery of clonogenic cells in suspension were considered to reflect well the self‐renewal of the clonogenic cells. When dose‐response curves were obtained for VCR and DOX, the primary clonogenic cells (PE1) were more sensitive than secondary clonogenic cells (PE2) or clonogenic cells in suspension. Repeated exposure to VCR in suspension did not inhibit the exponential growth of the clonogenic cells. These data indicated that both drugs were relatively ineffective in specifically suppressing the self‐renewal of the clonogenic cells.
Efficacy of Two‐route Chemotherapy Using Intraperitoneal Neocarzinostatin and Its Antidote, Intravenous Tiopronin, for Peritoneally Disseminated Tumors in Mice
Authors: Hasuda Keitaro , Kobayashi Hiroaki , Kuroiwa Toshiro , Aoki Ken , Taniguchi Shun'ichiro , Baba Tsuneo
Published: August 2005

We assessed the efficacy of “two‐route chemotherapy (TRC)” using neocarzinostatin (NCS) given ip and its antidote, N‐(2‐mercaptopropionyl)‐glycine (tiopronin), given iv for peritoneally disseminated tumors in mice. Whether or not the single iv administration of tiopronin (800 mg/kg) at various times after NCS ip would decrease the lethal toxicity induced by NCS ip was given attention. When compared with the LD50 (4.4 mg/kg) of NCS ip alone, simultaneous or postadministration of tiopronin together with NCS ip increased the LD50 of NCS ip by 2.8 to 7.6 fold in a time‐dependent manner. Chemotherapy experiments on ip disseminated tumors in mice were done to compare the antitumor effects of the following treatments, at two dose levels (75 and 100% of LD10) of NCS, with or without tiopronin: treatment with NCS ip alone and combined chemotherapy using NCS ip plus tiopronin iv, simultaneously or postadministered. Based on the survival time of the treated mice, the groups given NCS plus tiopronin (postadministration, 15 or 25 min later) showed a significantly superior survival time to that of the group given NCS ip alone. The side effects, evaluated in terms of the changes in body weight and number of WBC of the mice, were not significantly different among the groups treated with 100% of LD10 of NCS.
Leukocytosis in Mice Following Therapy with a Novel Antitumor Agent, RA‐700
Authors: Kato Taketoshi , Suzumura Yasuko , Liu Fen‐Zhi , Tateno Hiroo , Ogiu Toshiaki , Ota Kazuo
Published: August 2005

Nine daily intravenous (iv) injections of RA‐700 (an antitumor cyclic hexapeptide) at doses of 2 to 6 mgAg/day caused increases of WBC counts at 4–6 days after treatment in normal C57BL/6 × DBA/ 2 (BDF1) mice. The percentages of neutrophils and lymphocytes were modified. There was a decrease of colony‐forming units in culture (CFUc) in bone marrow to 40% of the control value on day 1 but CFUc rapidly returned to normal values on day 3. The colony‐forming units in spleen (CFUs): in bone marrow decreased during treatment. On the other hand, CFUc and CFUs in spleen were increased from the initiation of treatment to the time prior to the increase of WBC count. Spleen weight increased after treatment, and histologically, increases of immature and also mature granulocytes and megakaryocytes were observed. However, RA‐700 did not stimulate the progress of hematoprogenitors in vitro. The results indicated that RA‐700 stimulates the progress of hematoprogenitors in the spleen, but this effect is probably indirect.
An Epidemiological Study on Marital Status and Cancer Incidence
Authors: Kato Ikuko , Tominaga Suketami , Terao Chikahiro
Published: August 2005

The relationship between marital status and cancer incidence was examined based on 49,191 incident cases aged 30 or over in 1980–1984 by using the data from Aichi Cancer Registry and census data. Although married and widowed people did not show increased incidence for any cancer site studied, single and divorced people showed statistically significantly increased or decreased risks for several sites of cancer. Single males showed an increased risk for esophageal cancer and a decreased risk for lung cancer. Divorced males showed increased risks for cancers of the mouth & pharynx, esophagus, liver, skin and brain. Single females showed increased risks for cancers of the esophagus, stomach, small intestine, liver, pancreas, lung, breast, corpus uteri, ovary & fallopian tube and other female genital organs and a decreased risk for cervical cancer. Divorced females showed increased risks for cancers of the larynx, breast, all parts of uterus and cervix uteri and a decreased risk for biliary tract cancer. The increased risk for breast cancer in single females was more pronounced in older age groups and the increased risks for several sites of cancer in divorced people were more pronounced in younger age groups. These findings may be partly explained by differences in reproductive factors and life style, especially smoking and drinking habits.
Effects of Age on Multiple Organ Carcinogenesis Induced by 3,2′‐Dimethyl‐4‐aminobiphenyl in Rats, with Particular Reference to the Prostate
Authors: Shirai Tomoyuki , Nakamura Atsushi , Fukushima Shoji , Takahashi Satoru , Ogawa Kumiko , Ito Nobuyuki
Published: August 2005

The effects of age on multi‐organ carcinogenesis induced by 3,2′‐dimethyl‐4‐aminobiphenyl (DMAB) in male F344 rats were examined. Groups of 5‐, 35‐, and 65‐week‐old animals were given 4 weekly sc injections of DMAB at a dose of 200 or 150 mgAg body weight. Prostate carcinomas were induced in 8 to 19% of rats treated, no significant differences in the incidence between different ages being observed. Tumors in the small intestine, skin, pancreas and peritoneum, however, developed more frequently in young than in old animals, whereas higher incidences of testis, preputial and mammary gland lesions were found in the 35‐ and/or 65‐week‐old groups. Colon and Zymbal gland carcinogenesis did not reveal any age dependence.
Lack of Carcinogenicity of Quercetin in F344/DuCrj Rats
Authors: Ito Nobuyuki , Hagiwara Akihiro , Tamano Seiko , Kagawa Masataka , Shibata Masa‐Aki , Kurata Yasushi , Fukushima Shoji
Published: August 2005

Quercetin was administered at dietary levels of 0(control), 1.25 and 5.0% to groups of 50 male and 50 female rats for 104 weeks, and then all animals were maintained without quercetin supplement for a further 8 weeks. At 5.0% quercetin, both sexes showed growth retardation throughout the study. There were no treatment‐ascribed effects regarding clinical signs, mortality, urinalyses or hematology. Although serum glucose in 5.0% quercetin‐treated males was significantly decreased and some relative organ weights in 5.0% groups showed statistically significant increases, these latter changes seemed to be related to the growth retardation. An increased incidence of non‐neoplastic hyperplastic polyps in the cecum was noted in the 5.0% males. The incidences of cystic changes and fibroadenomas of the mammary gland, and foci (areas) of hepatocellular alteration in the 5.0% females, and liver bile duct proliferations in the 5.0% males were significantly decreased. No proliferative lesions of the urinary bladder related to treatment with quercetin were found in any rats. The incidences of several other nonneoplastic and neoplastic lesions which demonstrated statistically significant changes appeared to be related to the growth retardation or to be within the normal range, and therefore none was considered to be significant biologically. Thus, the investigation did not demonstrate any clear carcinogenic effect of quercetin on F344 rats at dietary levels of up to 5.0%.
Enhanced Liver Metastatic Potential of Alpha‐fetoprotein‐producing Human Gastric Carcinoma after Carbon Tetrachloride‐induced Liver Damage in Nude Mice
Authors: Sawada Hidetomo , Nakatani Katsunori , Miyagi Nobuyuki , Nishiwada Takashi , Watanabe Akihiko , Okumura Tohru , Yamada Yukishige , Tsutsumi Masahiro , Nakae Dai , Nakano Hiroshige , Konishi Yoichi
Published: August 2005

The liver metastatic potential of alpha‐fetoprotein (AFP)‐producing human gastric carcinoma (NSC‐3) was examined in male, BALB/c, nude mice. Metastatic nodules in the liver were produced by intrasplenic (IS) injection of tumor cell suspension prepared by trypsinization from subcutaneous NSC‐3 tumor. The serum AFP level increased exponentially after IS injection along with the growth of metastatic nodules in the liver, and a positive correlation was observed between the estimated weight of metastatic nodules and serum AFP level. To investigate the effect of liver damage by carbon tetrachloride (CCI4) on the metastatic potential of NSC‐3 cells injected intrasplenlcally, the mice were divided into 4 groups: Group 1 received IS injection of 1×106of NSC‐3 cells without CCI4, treatment; Groups 2, 3 and 4 received IS injection 7 days, 2 days and 1 day after CCI4treatment, respectively. All mice were killed 64 days after IS injection. The incidence of liver metastasis was 80% in Group 1, but 100% in Groups 2, 3 and 4. The mean numbers of metastatic nodules per liver were 4.2 in Group 1, 16.8 in Group 2, 18.0 in Group 3 and 44.5 in Group 4. Significant differences in the mean numbers of metastatic nodules were observed between Group 4 and the other groups. It was clearly demonstrated that the metastatic potential of AFP‐producing human gastric carcinoma cells (NSC‐3) is enhanced in the situation prevailing after liver parenchymal cells are damaged by CCI4.
Altered Expression of Epidermal Growth Factor Receptor Gene in a Classical Multidrug‐resistant Variant of a Human Cancer Cell Line, KB
Authors: Takano Hiroshi , Kohno Kimitoshi , Shiraishi Norio , Sato Shin‐ichi , Asoh Kuni‐ichi , Yakushinji Miki , Ono Mayumi , Kuwano Michihiko
Published: August 2005

A variant clone resistant to high doses of colchicine (KB‐C1) derived from human cancer KB cell line is resistant to various anticancer agents. The KB‐C1 cells were much more resistant to epidermal growth factor and a chimeric toxin, EGF‐Pseudomottas exotoxin (PE), than the parental KB cells. KB‐C1 cells have decreased numbers of EGF‐receptors, though the affinity of the receptors is similar to that in the parental KB cells. A drug‐sensitive revertant (C1‐R2) partially recovered its EGF‐receptor activity. Northern blot analysis showed a decreased level of EGF‐receptor mRNA in KB‐C1 cells, while the multidrug‐resistance gene, mdr‐1, was expressed at very high levels in KB‐C1 cells, but not in KB or C1‐R2 cells. The drug‐resistant cells were less tumorigenic than the parental cells when injected into nude mice. A decreased expression of EGF‐receptor in these cells may be one of the pleiotropic properties of multidrug‐resistant cells and may perhaps represent the basis for their reduced tumorigenicity.
Differential Neutralizing Effect of Tiopronin on the Toxicity of Neocarzinostatin and SMANCS: A New Rescue Cancer Chemotherapy
Authors: Oda Tatsuya , Yamamoto Hirotoshi , Miki Toyohiko , Maeda Hiroshi
Published: August 2005

The toxic effect and antitumor activity of neocarzinostatin (NCS) and SMANCS [copoly(styrenemaleic acid)‐conjugated NCS] were greatly affected by N‐(2‐mercaptopropionyl)‐glycine [tiopronm] both in vitro and in vivo, in cultured HeLa cells and RL♂1 tumor‐bearing mice. The cytotoxicity of NCS and SMANCS against HeLa cells was remarkably reduced by the addition of tiopronin during drug treatment. Interestingly, the neutralizing effect of tiopronin on the toxicity of SMANCS was greater than that in the case of NCS. In the continuous presence of 10 mM tiopronin during a 1 h drug treatment, the 50% cell‐killing doses of NCS and SMANCS were increased 72 and 208 times as compared to those without tiopronin, respectively, whereas tiopronin itself has no cytotoxicity to HeLa cells up to 100 mM. Furthermore, more effective reduction of the lethal toxicity of SMANCS was observed by the intraperitoneal (ip) administration of tiopronin after ip injection of a lethal dose of SMANCS as compared to the same protocol in the case of NCS in mice. Therapeutic studies on RL♂1 tumor‐bearing mice revealed that delayed (time lag) ip administration of tiopronin after high‐dose SMANCS administration ip was much superior to the combination of NCS with tiopronin, or SMANCS alone. In this time‐lag combination chemotherapy of SMANCS with tiopronin, 60% of treated mice survived more than 60 days after tumor inoculation, while all the untreated control mice died within 20 days.
Adult T‐cell Leukemia in Spouses
Authors: Sanada Isao , Yamaguchi Kazunari , Yoshida Mitsuaki , Ishii Toshinori , Tsukamoto Atsuko , Sato Masahiko , Obata Shin‐ichiro , Shido Tadahiro
Published: August 2005

A married couple who developed adult T‐cell leukemia (ATL) is described. The husband presented with acute ATL and died soon after admission in spite of aggressive chemotherapy, and his wife, who is diagnosed as smoldering ATL, has been followed in the out‐patient clinic. The couple had serum antibodies against human T‐cell leukemia virus type I (MTLV‐I) and the monoclonal integration of HTLV‐I proviral DNA in their lymphocytes. The patients described represent the first reported example of ATL in a married couple.
Quantitative Measurement of Cell Motility Associated with Transformed Phenotype
Authors: Tatsuka Masaaki , Jinno Shigeki , Kakunaga Takeo
Published: August 2005

The motility of individual mammalian cells is crucial for many biological processes. This report describes a new technique to quantitate cell motility momentary alterations of cell shape, based on trace images obtained by video‐image analyses and computer techniques. By means of this system, quantitation of cell motility could be automatically done without human observation or subjective judgement. Quantitative data from transformed and nontransformed rodent fibroblasts revealed that the cell motility measured here was related to the expression of such transformed phenotypes as morphological changes and tumorigenicity.
Oncogene Amplification in Squamous Cell Carcinoma of the Oral Cavity
Authors: Saranath Dhananjaya , Panchal Rekha G. , Nair Rama , Mehta Ashok R. , Sanghavi Vikram , Sumegi Janos , Klein George , Deo Madhav G.
Published: August 2005

We have determined the prevalence of amplification of c‐myc, N‐myc, L‐myc, H‐ras, Ki‐ras, and N‐ras oncogenes in 23 cases of squamous cell carcinoma of the oral cavity, using Southern hybridization analysis of DNA extracted from the primary tumor tissues. Nick‐translated oncogene probes and oncogene inserts labeled to high specific activities were used. We observed a 5‐ to 10‐fold amplification of one or more of c‐myc, N‐myc, Ki‐ras and N‐ras oncogenes in 56% of the tumor tissue samples, with these oncogenes not being amplified in the peripheral blood cells of the same patients, L‐myc and H‐ras were not amplified in any of our samples. The oncogene amplifications seemed to be associated with advanced stages of squamous cell carcinomas, with the ras and myc family oncogenes being amplified in stages 3 and 4. Hybridization with N‐myc detected an additional 2.3 kb EcoRI fragment, along with the normal 2.1 kb fragment. Our data also demonstrated amplification of multiple oncogenes in the same tumor tissue sample. About 60% of the samples with amplified oncogenes showed simultaneous amplification of 2 or more oncogenes. The results showing different oncogene amplifications in similar tumors, as well as multiple oncogene amplifications in the same tumor, suggest that these oncogenes may be alternatively or simultaneously activated in oral carcinogenesis.
Procoagulant Activity of Human Neuroblastoma Cell Lines, in Relation to Cell Growth, Differentiation and Cytogenetic Abnormalities
Authors: Esumi Noriko , Imashuku Shinsaku , Tsunamoto Kentaro , Todo Shinjiro , Misawa Shinichi , Goto Toshihiro , Fujisawa Yoshio
Published: August 2005

Procoagulant activity (PCA) was investigated in relation to cell growth, differentiation, and cytogenetics in seven human neuroblastoma cell lines. Before 5‐bromodeoxyuridine (BrdUrd) treatment, PCA was notably heterogeneous, with the highest activity in NCG (S‐type in morphology) 40‐to 100‐fold greater than the lowest activity in SK‐N‐D2 (N‐type). PCA was not related to 1p abnormalities. After BrdUrd treatment at 5 μg/ml for 6 days, PCA increased 6.8‐fold in GOTO and 2.7‐fold in SK‐N‐DZ with associated growth inhibition and morphological changes (I‐type morphology converted to S‐type in GOTO and N‐type converted to an advanced N‐type in SK‐N‐DZ). In contrast, only growth suppression was observed in 2 other cell lines, and no changes in PCA, growth or morphology were induced in the remaining 3 cell lines.
Unstable Expression of E‐Cadherin Adhesion Molecules in Metastatic Ovarian Tumor Cells
Authors: Hashimoto Miwa , Niwa Ohtsura , Nitta Yumiko , Takeichi Masatoshi , Yokoro Kenjiro
Published: August 2005

E‐Cadherin is a member of the cadherin family, which plays a key role in intercellular adhesion in various tumors as well as in normal tissues. Here, we examined the expression of this adhesion molecule in a murine ovarian tumor line OV2944, whose sublines show different degrees of spontaneous metastasis from subcutaneous sites; sublines LM‐1 and LM‐3 exhibit a low metastatic activity but a variant subline HM‐1 has a high metastatic activity. When the expression of E‐cadherin in these cells was examined by immunoblot analysis, the highly metastatic HM‐1 cells was found to express an extremely small amount of this molecule, as compared with a high level of E‐cadherin expression in the weakly metastatic LM‐1 and LM‐3 cells. Northern blot analysis showed that the amount of tanscripts from the E‐cadherin gene is proportional to the amount of proteins detected in these cells. Immunofluorescence staining revealed that cells of the highly metastatic line were heterogeneous, that is, their cultures contained both E‐cadherin‐positive and negative cells. In contrast, cells of the weakly metastatic lines homogeneously expressed E‐cadherin. When the highly metastatic line was subcloned, all the subclones consisted of E‐cadherin‐positive and negative cells. These results suggest that the expression of E‐cadherin gene is not stably controlled in the highly metastatic line.