bluuebunny/crossref_metadata_2025 huggingface dataset:
Poor Prognosis in Nasopharyngeal Cancer Patients with Low Glucose‐6‐phosphate‐dehydrogenase Activity
Authors: Cheng Ann‐Joy , Chiu Daniel Tsun‐Yee , See Lai‐Chu , Liao Chun‐Ta , Chen I‐How , Chang Joseph Tung‐Chieh
Published: August 2005

Nasopharyngeal carcinoma (NPC) is endemic among well‐defined ethnic groups in several world regions, such as Southeastern China and Taiwan. Glucose‐6‐phosphate‐dehydrogenase (G6PD)‐deficiency, a sex‐linked disorder, is one of the most common enzymopathies in Taiwan. The major role of G6PD is to generate NADPH to protect cells from oxidative damage, which is a major contributing factor to certain degenerative diseases, such as aging and cancer. In view of the coincidence of epidemic distribution of NPC and G6PD deficiency, as well as the house‐keeping function of G6PD in cellular oxidative defense, we investigated the correlation of G6PD activity with NPC. The stage of NPC was classified by AJCC (1997) criteria. G6PD levels were determined in 108 NPC male patients and 75 healthy male individuals. The mean G6PD level of NPC patients was 218.9 U/1012RBC or 7.53 U/g hemoglobin (Hb), being much lower than in normal individuals (260.6 U/1012erythrocytes (RBC) or 8.92 U/gHb). The level of G6PD activity had no correlation with tumor stage or lymph node or distant metastasis, but was significantly correlated with tumor recurrence (P=0.004 when using G6PD=130 U/1012 RBC as cutoff value). These results indicated that low G6PD activity in patients with NPC is associated with poor prognosis.
Association of ANA, a Member of the Antiproliferative Tob Family Proteins, with a Cafl Component of the CCR4 Transcriptional Regulatory Complex
Authors: Yoshida Yutaka , Hosoda Eri , Nakamura Takahisa , Yamamoto Tadashi
Published: August 2005

A 35‐kDa protein, ANA, belongs to an emerging family of antiproliferative proteins consisting of Tob, Tob2, ANA/BTG3, PC3B, PC3/TIS21/BTG2, and BTG1. All of these, except ANA and PC3B, have been shown to interact with the CCR4 transcription factor‐associated protein Cafl. Here we show that ANA also associates with Cafl, ANA being the preferred partner of Cafl among the Tob family proteins. Although ANA is likely to interact with Cafl at its amino‐terminal hall', which is conserved among the family members, our data suggest that the carboxyl‐terminal half of ANA plays a role in the interaction. Finally, in situ hybridization experiments revealed that expression of Caf1 overlaps at least in part with that of ANA. Thus, ANA could function through its interaction with Caf1.
Reversal of Cisplatin Resistance by the 1,4‐Benzothiazepine Derivative, JTV‐519
Authors: Nakamura Takashi , Koizumi Fumiaki , Kaneko Noboru , Tamura Tomohide , Chiwaki Fumiko , Koh Yasuhiro , Akutagawa Shigeru , Saijo Nagahiro , Nishio Kazuto
Published: August 2005

The 1,4‐benzothiazepine derivative JTV‐519 is a new type of calcium ion channel modulator. We examined the modulatory effect of JTV‐519 on the antitumor activity of several platinum compounds (cisplatin, carboplatin, and nedaplatin) in a human cancer cell line resistant to cisplatin (PC‐14/CDDP) in vitro. PC‐14/CDDP cells showed 8‐fold resistance to cisplatin compared with the parental PC‐14 cells as determined by dye formation [3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltet‐razolium bromide, MTT] assay. In PC‐14/CDDP, but not PC‐14 cells, augmentation of cytotoxicity was observed when a nontoxic concentration (10 μ) of JTV‐519 was combined with the platinum compounds. Increased intracellular cisplatin accumulation was observed in PC‐14/CDDP cells in the presence of JTV‐519 as measured by atomic absorption assay. Therefore, increased cisplatin accumulation was considered to be a possible mechanism underlying the reversing effect of JTV‐519 on cisplatin resistance. These results suggest that JTV‐519 is a potent agent reversing cisplatin resistance.
Chemopreventive Effects of Scordinin on Diethylnitrosamine and Phenobarbital‐induced Hepatocarcinogenesis in Male F344 Rats
Authors: Watanabe Tomoyuki , Sugie Shigeyuki , Okamoto Kiyohisa , Rahman K. M. Wahidur , Ushida Jun , Mori Hideki
Published: August 2005

Modifying effects of scordinin, a biological active component in garlic, on diethylnitrosamine (DEN)‐ and phenobarbital (PB)‐induced hepatocarcinogenesis were examined in rats. Male F344 rats, 5 weeks old, were divided into 8 groups. After a week, groups 1‐5 were given DEN (100 mg/ kg body weight, i.p.) once a week for 3 weeks, whereas groups 6‐8 received vehicle treatment. Group 2 was given 600 ppm scordinin‐containing diet in the initiation phase. From 4 weeks after the start of experiment, groups 3 and 5 were fed scordinin, and groups 1‐3 and 7 received drinking water containing 500 ppm PB. Group 6 was given scordinin diet alone throughout the experiment (24 weeks). The incidences of hepatocellular adenoma and carcinoma were significantly smaller in group 3 than those in group 1 (P<0.005 and P<0.05, respectively). The average numbers of liver neoplasms in groups 2 and 3 were significantly smaller than in group 1 (P<0.01 and P<0.0001, respectively). Glutathione S‐transferase placental form‐positive foci were also significantly decreased by scordinin treatment in the initiation or promotion phase. Scordinin significantly decreased the mean number of nucleolar organizer regions’ protein (AgNORs)/nucleus in hepatocellular adenoma and carcinoma. AgNORs/nucleus in the non‐lesional area was also significantly decreased by scordinin treatment during the promotion phase. These results suggest that scordinin is a promising chemopreventive agent for liver neoplasia.
Five Quantitative Trait Loci Affecting 4‐Nitroquinoline 1‐Oxide‐induced Tongue Cancer in the Rat
Authors: Tanuma Jun‐ichi , Fujii Kei , Hirano Masato , Matsuuchi Hiroaki , Shisa Hayase , Hiai Hiroshi , Kitano Motoo
Published: August 2005

In our previous study, Dark‐Agouti (DA) rats were found to be highly susceptible to 4‐nitroquino‐line 1‐oxide (4NQO)‐induced tongue carcinoma (TC), whereas Wistar/Furth (WF) rats were barely susceptible. Interval mapping analysis of reciprocal backcross rats showed two quantitative trait loci (QTL) on rat chromosomes (RNO) 1 and 19. In the present study, a composite interval mapping analysis was applied to 4NQO‐induced TC in 130 (DAxWF) F2 rats, demonstrating five independent QTL, Tongue squamous cell carcinoma 1‐5 (Tscc1‐5), responsible for phenotypic differences in the size and number of TCs in the two strains. Two of these QTL were mapped on RNO1, and the others were mapped on RNO4, 14, and 19. The DA allele at these loci consistently yielded semidominant susceptibility to TC. Out of the five loci detected in this F2 generation, Tscc1 and 2 were identical to Stcl and Rtc1 described in our previous study, but the other three were novel. We propose a new nomenclature consistent with their function. Genome‐wide screening of the F2 progeny also suggested the presence of three additional QTL on RNO5, 6, and 10. The possible roles of these loci in tongue carcinogenesis are discussed.
Ectopic Expression of MAFB Gene in Human Myeloma Cells Carrying (14;20)(q32;q11) Chromosomal Translocations
Authors: Hanamura Ichiro , Iida Shinsuke , Akano Yumiko , Hayami Yoshihito , Kato Miyuki , Miura Kazuhisa , Harada Shinsuke , Banno Shogo , Wakita Atsushi , Kiyoi Hitoshi , Naoe Tomoki , Shimizu Shiro , Sonta Shin‐ichi , Nitta Masakazu , Taniwaki Masafumi , Ueda Ryuzo
Published: August 2005

Chromosome 14q+, which represents a chromosomal rearrangement involving the immunoglobulin heavy chain gene (IgH) locus, is a genetic hallmark of human multiple myeloma (MM). Here, we report the identification of (14;20)(q32;qll) chromosomal translocations found in MM cells. Double color fluorescence in situ hybridization analyses pinpointed the breakpoints at the 20qll locus in two MM cell lines within a length of at most 680 kb between the KIAA0823 and MAFB gene loci. Among the transcribed sequences in the vicinity of the breakpoints, an ectopic expression of the MAFB gene, which is located at 450‐680 kb telomeric to one of the breakpoints and encodes a member of the MAF family basic region/leucine zipper transcription factor, was demonstrated to be associated with t(14;20). This finding, together with that of a previous study describing its transforming activity, suggests that the MAFB gene may be one of the targets deregulated by regulatory elements of the IgH gene as a result of t(14;20).
A Murine Osteosarcoma Cell Line with a Potential to Develop Ossification upon Transplantation
Authors: Kusumi Tomomi , Nishi Takashi , Tanaka Masanori , Tsuchida Shigeki , Kudo Hajime
Published: August 2005

An Osteosarcoma cell line has been established from a soft tissue tumor that occurred spontaneously in a BALB/c mouse. This cell line showed ossification when transplanted into syngeneic mice. To examine the mechanism of bone formation, the expression of mRNAs for osteoblastic and chon‐droblastic markers and factors associated with ossification has been investigated. In culture, the cells exhibited a spindle shape in the growth phase, but had a polygonal shape in the stationary phase. Reverse transcription‐polymerase chain reaction analysis showed that the cells expressed mRNAs for pro‐α(I) chain of type I collagen, alkaline phosphatase, osteopontin, osteocalcin, and core binding factor al, suggesting differentiation into the stage of osteoblasts during the stationary phase. After transplantation, histological examination revealed small foci of pale blue material and basophilic networks that were scattered in the tumor tissues at one week. The former stained positive with alcian blue, suggesting a chondroid matrix. Pro‐α(II) chain of type II collagen mRNA was expressed at one week. A large part of tumors at two and three weeks consisted of basophilic networks, which stained positive via von Kossa's method, indicating a calcified woven bone. In situ hybridization analysis showed strong expression of osteopontin and osteocalcin mRNAs in tumor cells surrounding the bone matrix. Bone morphogenetic protein‐6 and ‐7 mRNAs were detected in transplanted tumors, but not in cultured cells. These results suggest that the cell line has the properties of an osteoblastic lineage when cultured in vitro and has an ossifying ability through endochondral bone formation processes when transplanted in vivo.
Synergistic Effects of Highly Unsaturated Fatty Acid‐containing Phosphatidyl‐ethanolamine on Differentiation of Human Leukemia HL‐60 Cells by Dibutyryl Cyclic Adenosine Monophosphate
Authors: Hosokawa Masashi , Sato Akiyuki , Ishigamori Hideki , Kohno Hiroyuki , Tanaka Takuji , Takahashi Koretaro
Published: August 2005

Highly unsaturated fatty acid‐containing phospholipid (HUFA‐PL) has many nutritional and medical applications. We investigated the effect of HUFA‐PL on differentiation of human leukemia HL‐60 cells induced by dibutyryl cyclic adenosine monophosphate (dbcAMP). HUFA‐containing phosphatidylethanolamine (HUFA‐PE), such as salmon testis PE, significantly enhanced dbcAMP ‐induced cell differentiation. A combined treatment of 200 μM dbcAMP with 50 μM HUFA‐PE increased the nitroblue tetrazolium (NBT)‐reducing activity, which is an indicator of differentiation, to a level comparable to that in the case of 500 μM dbcAMP treatment. In contrast, HUFA‐lyso PE (a monoacyl form) did not exert an enhancing effect on dbcAMP‐induced differentiation. The enhancing effect of HUFA‐PE was suppressed by a protein kinase C inhibitor, staurosporine, while a protein kinase A inhibitor, H‐8, did not suppress the enhancing effect. These findings suggest that HUFA‐PE might enhance dbcAMP‐induced differentiation through modulation of the protein kinase C signaling pathway in HL‐60 cells.
Sequential Histopathological Changes in vivo after Suicide Gene Therapy of Gastric Cancer Induced by N‐Methyl‐N′‐nitro‐N‐nitrosoguanidine in Rats
Authors: Okino Tetsuya , Onda Masahiko , Matsukura Norio , Inada Ken‐iti , Tatematsu Masae , Suzuki Satoru , Shimada Takashi
Published: August 2005

Gastrointestinal cancer is the most important clinical target of gene therapy. Suicide gene therapy, such as with the herpes simplex virus type 1 thymidine kinase (HSV‐TK) gene, has been shown to exert antitumor efficacy in various cancer models in vitro. We previously reported in situ gene transfer and gene therapy for gastric cancer induced by N‐ethyl‐.N′‐nitro‐.N‐nitrosoguanidine (ENNG) in dogs. Here, we describe the sequential histopathological changes after suicide gene therapy of N‐methyl‐.N′‐nitro‐.N nitrosoguanidine (MNNG)‐induced gastric cancer in rats. Gastric tumors were induced by MNNG in 38/73 (52%) of Wistar strain rats. The suicide gene therapy group (14 rats) was subjected to in situ gene transfer with a recombinant adenovirus vector carrying the HSV‐TK gene driven by CAG promoter (Ad.CAGHSV‐TK) in gastric tumor, followed by the antiviral drug ganciclovir (GCV). To observe the histopathological changes at various tunes after HSV‐TK/GCV gene therapy, groups of animals were sacrificed at 3, 8, and 30 days after gene transfer. Apoptosis in the gastric tumors was detected by the TUNEL method to assess the efficacy of HSV‐TK/GCV gene therapy, and it was marked in the 8‐ and 30‐day treatment groups compared to the sham operation controls (P<0.001). Various histopathological changes, degeneration of cancer tissue and fibrosis after necrosis and apoptosis were significantly greater in the 30‐day treatment group. The HSV‐TK gene was detectable in peripheral blood by PCR until 30 days after gene transfer. These results may be useful in devising a method of suicide gene therapy for humans.
Effect of Combination Therapy with Matrix Metalloproteinase Inhibitor MMI‐166 and Mitomycin C on the Growth and Liver Metastasis of Human Colon Cancer
Authors: Ohta Manabu , Konno Hiroyuki , Tanaka Tatsuo , Baba Megumi , Kamiya Kinji , Oba Kouji , Kaneko Takeshi , Syouji Tsuyoshi , Igarashi Akira , Nakamura Satoshi
Published: August 2005

Several synthetic inhibitors of matrix metalloproteinases (MMPs) show antitumor, antimetastasis and antiangiogenesis effects in various models. Synergistic effects of combinations with conventional cytotoxic agents were reported previously. In this study, we examined the effects of a new selective MMP inhibitor, MMI‐166, on tumor growth, angiogenesis and metastasis in a liver meta‐static model of human xenotransplanted colon cancer (TK‐4). We also investigated the synergistic effects of MMI‐166 and a conventional cytotoxic agent, mitomycin C (MMC), in this model. Mice transplanted orthotopically with TK‐4 were divided into 4 groups; a control group (treated with vehicle solution), an MMI‐166 group in which MMI‐166 was orally administered (p.o.) at a dose of 200 mg/kg, 6 days/week for 5 weeks, an MMC group in which MMC was administered intraperitoneally (i.p.) at a dose of 2 mg/kg/week for 5 weeks, and a combination group (treated with MMI‐166 and MMC). MMI‐166 did not inhibit transplanted tumor growth, but significantly inhibited liver metastasis compared with the control group and MMC group (P<0.01). Significant antitumor and antimetastatic effects of the combination therapy were demonstrated. The microvessel density (MVD) detected by immunohistochemical staining with ER‐MP12 antibody tended to be lower in the MMI‐166 and the combination groups. These results suggest that MMI‐166 has potential antimetastatic ability and a synergistic effect with MMC.
DNA Sequence Copy Number Aberrations Associated with Histological Subtypes and DNA Ploidy in Gastric Carcinoma
Authors: Kong Gu , Oga Atsunori , Park Chan Kum , Kawauchi Shigeto , Furuya Tomoko , Sasaki Kohsuke
Published: August 2005

We have analyzed DNA sequence copy number aberrations (DSCNAs) and DNA ploidy by using comparative genomic hybridization and laser scanning cytometer in gastric carcinomas (GCs) to elucidate the genomic aberrations in relation to clinicopathological parameters. Thirty‐two out of 33 cases showed one or more DSCNAs with a mean number of 11.7 per tumor. High‐level gains were detected at 2p, 3q, 6p, 7p, 7q, 8q, 12p, 13q, 19q, and 20q. Frequency of gross genomic abnormalities and chromosome regions that have genomic aberrations were similar in both intestinal‐and diffuse‐type GCs, except aberrations at 8p, 9p, 12q, and 20q. The overall number of DSCNAs was significantly greater in DNA aneuploid tumors than that in DNA diploid tumors. We detected genomic aberrations characterized by histological subtype, tumor location, and DNA ploidy status: gain of 20q and losses of 8p and 9p in intestinal‐type GCs, gains of 8p and 12q in diffuse‐type GCs, gain of 20q in the lower third GCs, and loss of 5q, 9p, lOq, 16q, and 18q in DNA aneuploid GCs. Furthermore, 5q loss is associated with DNA aneuploidy (P=0.0001) or the total number of losses (P=0.001), gain+losses (P=0.004), and high‐level gains (P=0.001) in GCs. Among these loci, chromosome 8p was unique. Gain of 8p was more common in diffuse‐type GC, whereas loss of 8p was more frequently detected in intestinal‐type GC. In conclusion, we describe chromosomal regions of 5q, 8p, and 20q, which are of interest for further investigation of GCs.
Hypophosphorylation of Topoisomerase IIα in Etoposide (VP‐16)‐resistant Human Carcinoma Cell Lines Associated with Carboxy‐terminal Truncation
Authors: Matsumoto Yoshihito , Takano Hiroshi , Kunishio Katsuzo , Nagao Seigo , Fojo Tito
Published: August 2005

Topoisomerase Ilα is a target for many chemotherapeutic agents in clinical use. To define mechanisms of resistance and regions crucial for the function of topoisomerase IIα, drug‐resistant cell lines have been isolated following exposure to topoisomerase II poisons. Two resistant sublines, T47D‐VP and MCF‐7‐VP, were isolated from human carcinoma cell lines following exposure to 300 or 500 ng/ml etoposide (VP‐16). Cytotoxicity studies confirmed resistance to etoposide and other topoisomerase II poisons. KCl‐sodium dodecyl sulfate (K‐SDS) precipitation assays using intact cells showed reduced DNA‐topoisomerase II complex formation following VP‐16 or amsacrine (m‐AMSA). RNAse protection analysis identified a deletion of 200 base pairs in the topoisomerase Ilα cDNA of T47D‐VP and “AA insertion” in the topoisomerase Ilα cDNA of MCF‐7‐VP. Reduced topoisomerase Ila mRNA and protein levels were observed in both cell lines. It was somewhat surprising to find that nuclear extracts from T47D‐VP and MCF‐7‐VP cells had comparable topoisomerase II activity to that of parental cells. Analysis of the extent of phosphorylation demonstrated that topoisomerase Ila from the resistant cells was relatively hypophosphorylated compared to that of parental cells. In these cell lines, hypophosphorylation secondary to loss of a portion of the C‐terminal domain of topoisomerase Ilα mediated the restored activity, despite a fall in topoisomerase Ila mRNA and protein, and this resulted in cross resistance to topoisomerase II poisons.
Epstein‐Barr Virus‐associated Gastric Carcinoma in Japanese Brazilians and Non‐Japanese Brazilians in São Paulo
Authors: Koriyama Chihaya , Akiba Suminori , Iriya Kiyoshi , Yamaguti Thiana , Hamada Gerson S. , Itoh Tetsuhiko , Eizuru Yoshito , Aikou Takashi , Watanabe Shaw , Tsugane Shoichiro , Tokunaga Masayoshi
Published: August 2005

The proportion of Epstein‐Barr virus‐associated gastric carcinoma (EB V‐GC) was examined in 149 Japanese‐Brazilian and 151 non‐Japanese‐Brazilian gastric‐carcinoma cases usingin situhybridization (ISH) assay to detect EBV‐encoded small RNA (EBER), and the results were compared with our referent Japanese data. We found that 4.7% of cases in Japanese Brazilians were EBER‐positive. This frequency was slightly lower than that of the referent Japanese, among whom 6.2% of 2038 gastric‐carcinoma cases were EBER‐positive. On the other hand, the non‐Japanese‐Brazilian series showed a significantly higher proportion of EBV‐GC (11.2%) than the referent group did (P=0.01). Although EBV‐GC was predominant in males among non‐Japanese Brazilians (M/ F=3.6, P=0.047), as was the case in Japanese (M/F=2.7), Japanese Brazilians did not show such a male predominance. The sex‐ratio difference between the Japanese Brazilians and Japanese was statistically significant (P=0.005). In conclusion, the present study in Japanese Brazilians and Japanese yielded no evidence suggesting any change in the frequency of EBV‐GC caused by migration, except the absence of male predominance, which was observed both in Japanese and non‐Japanese Brazilians.
Mitotic Checkpoint Protein hsMAD2 as a Marker Predicting Liver Metastasis of Human Gastric Cancers
Authors: Tanaka Kouji , Nishioka Junji , Kato Keiko , Nakamura Akiko , Mouri Tomomi , Miki Chikao , Kusunoki Masato , Nobori Tsutomu
Published: August 2005

hsMAD2, the human homologue of mitotic arrest deficient 2 (MAD2), is a key component of the mitotic checkpoint system. Recently, mutations and decreased expression of mitotic checkpoint genes including hsMAD2 have been reported in cancer cell lines with defective mitotic checkpoint. However, the genetic alterations in the genomic hsMAD2 gene have not been determined in gastric cancers. Moreover, the biological implications of the overexpressed hsMAD2 in primary cancers are unknown. In this study, we analyzed 32 primary gastric cancers with polymerase chain reaction (PCR) amplification of all exons, including flanking intronic sequences, of the genomic hsMAD2 gene followed by direct DNA sequencing. We also measured the hsMAD2 protein levels in cancer and normal tissues by semi‐quantitative immunoblotting. No mutations were found in the coding sequences, although three single nucleotide polymorphisms (SNPs) were identified in the noncoding sequences in 13 of 32 patients. These SNPs were not associated with either hsMAD2 expression or disease progression. The semi‐quantitative western blot analysis showed hsMAD2 was significantly overexpressed in gastric cancer tissues compared with corresponding normal tissues (P<0.001). The calculated ratio of the hsMAD2 protein in cancer tissue (C) to that in corresponding normal tissue (N) (C/N ratio) was significantly higher in patients with well differentiated adenocarcinoma (P=0.0274) or with synchronous liver metastasis (P=0.0025). A C/N ratio greater than 3 was observed more frequently in patients with synchronous liver metastasis. Therefore, C/N ratio >3 may be clinically important as a predictive indicator for metachronous liver metastasis of gastric cancers.
Pharmacokinetic Study of a Gallium‐porphyrin Photo‐ and Sono‐sensitizer, ATX‐70, in Tumor‐bearing Mice
Authors: Sasaki Kazuaki , Yumita Nagahiko , Nishigaki Ryuichiro , Sakata Isao , Nakajima Susumu , Umemura Shin‐ichiro
Published: August 2005

The tissue distribution of a gallium‐porphyrin photo‐ and sono‐sensitizer, 7,12‐bis(l‐decyloxy‐ethyl)‐Ga(III)‐3,8,13,17‐tetramethylporphyrin‐2,18‐dipropionyldiaspartic acid, ATX‐70, was phar‐niacokinetically examined in tumor‐bearing mice. The drug was administered intravenously to CDF1mice implanted with Colon 26 carcinoma. Blood and tissue samples were collected for up to 72 h after administration. The drug concentration was determined by high‐performance liquid chromatography (HPLC) with fluorescence detection. ATX‐70 was found to accumulate in tumors at a relatively high concentration that peaked between 2 h and 6 h after administration. However, modest concentrations of ATX‐70 also remained in healthy tissues for up to 6 h. We examined the distribution of ATX‐70 in the tumor in comparison with other tissues from the viewpoint of minimizing possible side effects of laser or ultrasound exposure while maintaining the treatment effect. About 24 h after administration, the tumor/plasma concentration ratio peaked, and relatively high tumor/skin and tumor/muscle concentration ratios were seen.
Organotropic Chemopreventive Effects of n‐3 Unsaturated Fatty Acids in a Rat Multi‐organ Carcinogenesis Model
Authors: Toriyama‐Baba Hiroyasu , Ligo Masaaki , Asamoto Makoto , Iwahori Yoshio , Park Cheol Beom , Han Beom Seok , Takasuka Nobuo , Kakizoe Tadao , Ishikawa Chikako , Yazawa Kazunaga , Araki Eiji , Tsuda Hiroyuki
Published: August 2005

Organotropic chemopreventive effects of n‐3 unsaturated fatty acids were studied using a multi‐organ carcinogenesis model in male rats. Rats were treated with diethylnitrosamine (DEN), N‐methyl‐N‐nitrosourea (MNU), N‐butyl‐N‐4‐hydroxybutylnitrosamine (BBN), 1,2‐dimethylhydrazine (DMH) and dihydroxy‐di‐n‐propylnitrosamine (DHPN) during the first 7 weeks, and then given unsaturated fatty acid (UFAs), docosahexaenoic acid (n‐3, C22:6) (DHA), eicosapentaenoic acid (n‐3, C20:5) (EPA), linoleic acid (n‐6, C18:2) (LA) or oleic acid (n‐9, C18:1) (OA) at a dose of 1.0 ml/rat, 3 tunes a week by gavage for the consecutive 30 weeks. All rats were fed a low LA basal diet throughout the experiment and a calorie‐restricted basal diet during the period of UFAs feeding administration. DHA significantly reduced tumor size and numbers in the large intestine as compared to OA treatment. Furthermore, DHA showed a tendency to inhibit carcinogenesis in the small intestine and lung. EPA also showed a tendency to inhibit intestinal carcinogenesis. On the other hand, LA showed a tendency to inhibit lung carcinogenesis, but to promote large intestinal carcinogenesis. However these UFAs did not influence preneoplastic and neoplastic lesion development in the liver, kidney, and urinary bladder. Levels of the administered fatty acids were clearly increased in the serum and organs. In contrast, arachidonic acid (AA) levels in the large and small intestines and liver were markedly decreased by treatment with DHA and EPA. Decreased levels of AA in the large intestine correlated well with tumor incidence, although the number of glutathione S‐transferase‐positive (GST‐P+) foci showed an inverse correlation with AA levels. The data thus provide evidence that an organotropism exists with regard to the influence of UFAs on carcinogenesis, which correlates with reduction of tissue AA levels in the target organs.
High Frequency of Deletions at the Hypoxanthine‐guanine Phosphoribosyl‐transferase Locus in an Ataxia‐telangiectasia Lymphoblastoid Cell Line Irradiated with y‐Rays
Authors: Tachibana Akira , Tatsumi Kouichi , Furuno‐Fukushi Ikuko , Sasaki Masao S.
Published: August 2005

The molecular nature of y‐ray‐induced mutations at the hypoxanthine‐guanine phosphoribosyl‐transferase (HPRT) locus in an ataxia‐telangiectasia (A‐T) lymphoblastoid cell line was investigated. Twelve of 15 y‐ray‐induced HPRT‐deficient mutants showed deletions. Eight of them had lost the entire HPRT gene, one showed a 1.9‐kb deletion, and three had deletions of about 40–150 base pairs. Of the eight mutants that lost the entire gene, five had also lost both DXS79 and DXS86, flanking markers of the HPRT locus. The spectrum of mutations induced by y‐irradiation in the A‐T cells showed a high frequency of deletions in comparison with that in a control cell line, WIL2NS. Sequence analysis of breakpoint junctions in four mutants revealed that three of them had junctions between short identical sequences at each breakpoint, leaving one copy at the junction. These results suggest that non‐homologous end‐joining is the major mechanism for deletion formation in A‐T cells.
Association of Allelic Losses at 3p25.1, 13q12, or 17p13.3 with Poor Prognosis in Breast Cancers with Lymph Node Metastasis
Authors: Haga Shunsuke , Emi Mitsuru , Hirano Akira , Utada Yoshihito , Kajiwara Tetsuro , Akiyama Futoshi , Sakamoto Goi , Takahashi Kaoru , Tada Takashi , Kasumi Fujio , Miki Yoshio , Nakamura Yusuke
Published: August 2005

To identify specific allelic losses that might correlate with postoperative mortality of patients with node‐positive breast carcinomas, we examined tumors from a cohort of 263 such patients, who were followed clinically for 5 years postoperatively, for allelic losses among 18 microsatellite markers. Patients whose tumors had lost an allele at 3p25.1, 13q12, or 17p13.3 had significantly higher risks of mortality than those whose tumors retained both alleles at those loci. At 3p25.1, the 5‐year mortality rate was 33.8% among patients with losses vs. 16.8% with retention (P=0.0154); at 13q12,30.3% vs. 13.0% (P=0.0241); and at 17p13.3, 30.4% vs. 16.2% (P=0.0243). Combined losses at 3p25.1 and 17p13.3 increased the predicted postoperative mortality risk by a factor of 4.9 (5‐year mortality rate of 38.2% vs. 8.0%, P=0.0006), and combined losses at 3p25.1 and 13q12 raised the predicted postoperative mortality risks by a factor of 2.9 (34.7% vs. 12.7%, P=0.0441). These data indicate that loss of heterozygosity (LOH) at any one or a pair of loci at 3p25.1, 13q12, or 17p13.3 is a significant predictor of postoperative mortality for breast‐cancer patients.
Macrophage Colony‐stimulating Factor Prevents Febrile Neutropenia Induced by Chemotherapy
Authors: Hidaka Takao , Fujimura Masaki , Sakai Masatoshi , Saito Shigeru
Published: August 2005

There are very few studies describing the preventive effect of macrophage colony‐stimulating factor (M‐CSF/CSF‐1) on chemotherapy‐induced infection. In this study, we evaluated the changes in superoxide anion production by granulocytes before and after chemotherapy in ovarian cancer patients and investigated the preventive effect of M‐CSF on chemotherapy‐induced febrile neutropenia. Three courses of chemotherapy [paclitaxel 180 mg/m2 and carboplatin (area under the curve; AUC 5)] were administered to 32 ovarian cancer patients, and seven patients presented febrile neutropenia. In the 25 afebrile patients, the percentage of superoxide anion production by granulocytes was significantly decreased from 86.5±7.7 (%) to 75.1±8.8 (%) at day 7 and 71.0±6.3 (%) at day 14 without administration of CSF. However, in the patients who presented febrile neutropenia, it was more severely decreased from 86.8±6.8 (%) to 60.0±9.9 (%) at day 7 and 56.8±5.0 (%) at day 14 without administration of CSF. When M‐CSF was administered to all patients in the next course with the same dose of chemotherapy, the incidence of febrile neutropenia was significantly decreased (P=0.0195), and the duration of fever (≫38.0°C) and high serum C‐reactive protein (CRP) (≫2.0 mg/dl) were also significantly shortened (P=0.0023, P=0.0051). Moreover, in these M‐CSF‐treated patients, the percentage of superoxide anion production by granulocytes was maintained at the level before chemotherapy. These findings indicate that severe impairment of granulocyte function leads to febrile neutropenia, and that M‐CSF reduces the incidence of febrile neutropenia by maintaining or improving granulocyte function.
Dietary Habits and Risk of Lung Cancer Death in a Large‐scale Cohort Study (JACC Study) in Japan by Sex and Smoking Habit
Authors: Ozasa Kotaro , Watanabe Yoshiyuki , Ito Yoshinori , Suzuki Koji , Tamakoshi Akiko , Seki Nao , Nishino Yoshikazu , Kondo Takaaki , Wakai Kenji , Ando Masahiko , Ohno Yoshiyuki ,   JACC Study Group
Published: August 2005

Lung cancer has increased and is the leading cause of cancer death among Japanese males. The associations of dietary habits with the risk of lung cancer death were evaluated by sex and smoking habits in this study. In the Japan Collaborative Cohort (JACC) Study, a cohort established in 1988–90 and consisting of 42 940 males and 55 308 females was observed for lung cancer deaths up to the end of 1997. During the observation period, 446 males and 126 females died of lung cancer. A self‐administered food frequency questionnaire was used as the baseline survey. Hazard ratios for dietary factors were calculated by Cox's proportional hazards model. Among males, a high intake of ham and sausages, cheese, green‐leafy vegetables, oranges, and other fruits significantly and dose‐dependently decreased the risk of lung cancer death. Among females, a high intake of miso‐soup, ham and sausages, and liver significantly and almost dose‐dependently increased the risk. Vegetables and fruits rich in antioxidative and carcinogenic agents reduced the risk of lung cancer deaths among male smokers more than among female nonsmokers. The results among female nonsmokers were partially consistent with the hypothesis that high fat consumption increases the risk of lung cancer, especially that of adenocarcinoma.
Aberrations of the p14ARF and p16INK4a Genes in Renal Cell Carcinomas
Authors: Kawada Yoichi , Nakamura Mitsutoshi , Ishida Eiwa , Shimada Keiji , Oosterwijk Egbert , Uemura Hirotsugu , Hirao Yoshihiko , Chul Kim Sung , Konishi Noboru
Published: August 2005

The INK4alARF locus on chromosome 9p21, which encodes two distinct genes, p14ARF and p16INK4a, is frequently altered in human neoplasms. To investigate the potential roles of p14ARF and p16IfiK4a genes in human renal cell carcinomas (RCCs), we analyzed 6 human RCC cell lines and 91 primary RCCs for homozygous deletion, promoter hypermethylation and expression of the p14ARF and p16INK4a gene products using differential PCR, methylation‐specific PCR, and immunohistochemis‐try, respectively. Five cell lines showed homozygous co‐deletion of both genes and one demonstrated promoter hypermethylation of the p16INK4a gene only. Eight of 91 RCCs showed aberrations of p14ARF or p16INK4a status and six of these featured gross extension into the renal vein. The results suggest that p14ARF and p16INK4a aberrations may play roles in the relatively late stage of renal tumorigenesis associated with tumor progression.
Expression Profile of Histone Deacetylase 1 in Gastric Cancer Tissues
Authors: Choi Jae‐Hoon , Kwon Ho Jeong , Yoon Byung‐II , Kim Jin‐Hyun , Han Sang Uk , Joo Hee Jae , Kim Dae‐Yong
Published: August 2005

Although histone deacetylases (HDACs) appear to play a crucial role in carcinogenesis, the expression status of HDACs in primary human cancer tissues has not yet been reported. In this study, we investigated the expression level of HDAC1 in 25 paired primary human gastric cancer (GC) tissues and corresponding normal tissues through semi‐quantitative RT‐PCR and immunoblot analysis. The HDAC1 expression pattern was also topologically examined through immunohisto‐chemistry. Overexpression of HDAC1 mRNA was detected in 68% of GC tissues (17 of 25), and the relative density of HDAC1 mRNA in GC tissue was increased 1.8‐fold versus the normal counterpart (P<0.01). Elevated expression of HDAC1 protein was also detected in 61% of GC samples (11 of 18), which also showed an increased mRNA level of HDAC. Immunohistochemically, overexpression of HDAC1 was predominantly localized in the nuclei of most neoplastic cells, including embolic tumor cells, whereas normal glandular epithelial cells revealed only weak HDAC1 expression that was focal in distribution. Thus, the present study clearly demonstrates that HDAC1 is overexpressed in GC and probably plays a significant role in gastric carcinogenesis.
Clonality Assay of Hematopoietic Disorders: Significance of the Buccal Epithelium as Non‐hematopoietic Control and of 95% Rejection Limit as a Novel Criterion for Monoclonality
Authors: Inagaki Hiroshi , Wakita Atsushi , Komatsu Hirokazu , Kikuchi Motoo , Inagaki Aki , Eimoto Tadaaki , Ueda Ryuzo
Published: August 2005

In clonality assays using × chromosome inactivation patterns (XCIPs), several factors such as constitutive and acquired XCIP skewing, lack of appropriate controls for hematopoietic diseases including multilineage disorders, and ambiguous criteria for monoclonality, have complicated determination of clonality. To address these issues, we studied the significance of the buccal epithelium as a non‐hematopoietic control and the usefulness of the 95% rejection limit as a criterion for monoclonality. Sixty‐nine females informative for human androgen receptor gene (HUMARA) were divided into “young,”“middle‐aged” and “elderly” groups. When XCIP correlation between the buccal epithelium, peripheral granulocytes, and peripheral lymphocytes was analyzed, the buccal epithelium showed a good correlation with granulocytes and lymphocytes in “young” and “middle‐aged” groups, whereas the correlation was poor for the “elderly” group. For all age groups, there was an excellent correlation between granulocytes and lymphocytes. When we performed clonality assay for seven “young” and “middle‐aged” patients with various leukemic phases using buccal epithelium as a non‐hematopoietic control, all cases were accurately evaluated with the aid of a novel criterion, the 95% rejection limit. Our findings suggest that the buccal epithelium may constitute an effective control, especially when a non‐hematopoietic control is required, and that the 95% rejection limit may serve as a statistically‐defined criterion for monoclonality.
Phage Display Cloning and Characterization of Monoclonal Antibody Genes and Recombinant Fab Fragment against the CD98 Oncoprotein
Authors: Itoh Kunihiko , Inoue Kazuyuki , Hirooka Kazumi , Maruyama Kumiko , Ohkawa Mayumi , Matsui Kazuhiro , Tada Hitoshi , Enomoto Takemi , Hashimoto Yoshiyuki , Suzuki Toshio , Masuko Takashi
Published: August 2005

The Fab gene of anti‐CD98 heavy chain (h.c.) monoclonal antibody (mAb) HBJ127 was cloned and expressed as a recombinant Fab (rFab) fragment by means of a phage display system. The variable heavy and light chain genes of HBJ127 were found to be derived from VOx‐1 and IgVk8‐30 germline, respectively. Extensive somatic mutation was found in the heavy chain complementarity determining region 2. rFab fragment was purified homogeneously from crude bacterial lysates by Ni‐chelate chromatography in a yield of 71.4 μg from 100 ml of culture. rFab fragment was reactive with the cell surface of CD98‐positive cells irrespective of tissues of origin, but not with CD98‐neg‐ative cells. The recognition site of the rFab fragment was identical to that of mAb since the binding of rFab fragment to HeLaS3cells was completely inhibited by pretreatment with an excess of mAb. The relative affinity values of rFab fragment and mAb were found to be 0.11x108 and 0.35X108M‐1, respectively. Three‐fold lower affinity of rFab fragment may be due to the difference of valency of the antibody preparation. Cell growth inhibition in vitro by rFab fragment preincubated with anti‐Fab suggests that the rFab fragment produced by cloned gene‐bearing Escherichia coli was identical to the Fab part of HBJ127 mAb. These results show that a small fragment with antigen binding activity similar to that of the parent mAb can easily be prepared by using a phage display system. To our knowledge, this is a first report of the production of anti‐CD98 h.c. rFab fragment.
The Citrus Flavonoid, Nobiletin, Inhibits Peritoneal Dissemination of Human Gastric Carcinoma in SCID Mice
Authors: Minagawa Akiko , Otani Yoshihide , Kubota Tetsuro , Wada Norihito , Furukawa Toshiharu , Kumai Koichiro , Kameyama Kaori , Okada Yasunori , Fujii Masato , Yano Masamichi , Sato Takashi , Ito Akira , Kitajima Masaki
Published: August 2005

The flavonoid nobiletin (5,6,7,8,3′,4′–hexamethoxyflavone), found in Citrus depressa Rutaceae, a popular citrus fruit in Okinawa, Japan, reportedly inhibits the production of pro–matrix metallo–proteinase (proMMP)–l, 3, and 9 in rabbit synovial fibroblasts in vitro. In the present study, we demonstrated the inhibitory effects of nobiletin on the proliferation of the cancer cell line, TMK–1, and its production of MMPs. In the SCID mouse model, we found that nobiletin inhibited the formation of peritoneal dissemination nodules from TMK–1. The enzymatic activity of MMP–9 expressed in culture medium obtained from a co–culture of TMK–1 and mouse fibroblastic cells was inhibited by nobiletin in a concentration–dependent manner. In the SCID mouse model, total weight of dissemination nodules was significantly lower in the treated group compared with the vehicle control group (0.07 g vs. 0.78 g, P=0.0059). The total number of dissemination nodules was also significantly lower than in the vehicle control group (7.5 vs. 69.3/body, P=0.0001). These results suggest that nobiletin may be a candidate anti–metastatic drug for prevention of peritoneal dissemination of gastric cancer.
L‐myc Genotype is Associated with Different Susceptibility to Lung Cancer in Smokers
Authors: Kumimoto Hiroshi , Hamajima Nobuyuki , Nishimoto Yoshio , Matsuo Keitaro , Shinoda Masayuki , Hatooka Shunzo , Ishizaki Kanji
Published: August 2005

We have shown that L‐myc genotype is associated with the risk of esophageal cancer from smoking and heavy drinking. In this study, we have analyzed the relationship between the L‐myc genotypes and lung cancer risk from smoking in 191 Japanese lung‐cancer patients and 241 non‐cancer controls. The odds ratios (ORs) were markedly higher in SS and LS genotypes than in LL genotype; age‐sex‐adjusted ORs were 3.19, 2.30 and 0.92, respectively. This result suggests that the L‐myc polymorphism may affect the induction of lung cancer by smoking. The OR for smoking in SS‐genotype patients diagnosed within 2 years was higher than that in other SS patients, suggesting that smoking‐related lung cancer in SS genotype might exhibit a poorer prognosis.
Enhancement by Cigarette Smoke Exposure of 2‐Amino‐3,8‐dimethylimidazo[4,5‐ f]quinoxaline‐induced Rat Hepatocarcinogenesis in Close Association with Elevation of Hepatic CYP1A2
Authors: Nishikawa Akiyoshi , Furukawa Fumio , Miyauchi Makoto , Son Hwa‐Young , Okazaki Kazushi , Koide Akihiro , Mori Yukio , Hirose Masao
Published: August 2005

The modifying effects of cigarette smoke (CS) exposure on a heterocyclic amine (HCA) 2‐amino‐3,8‐dimethylimidazo[4,5‐f]quinoxaline (MeIQx)‐induced carcinogenesis were investigated in male F344 rats. Groups 1 and 2 were fed MeIQx at a dose of 300 ppm, and simultaneously received CS and sham smoke (SS) for 16 weeks, respectively. Groups 3–5 were given the MeIQx diet for 4 weeks, and simultaneously exposed to CS for 4 weeks (group 3), exposed to CS for 12 weeks after the MeIQx treatment (group 4) or received SS for 16 weeks (group 5). Groups 6 and 7 were fed basal diet and respectively received CS and SS for 16 weeks. In terms of the mean number or area, the development of glutathione S‐transferase placental form‐positive (GST‐P+) liver cell foci was significantly (P<0.01) greater in group 1 than in group 2. The mean number of colonic aberrant crypt foci (ACFs) per animal was increased by continuous CS exposure regardless of MeIQx feeding, the differences between groups 4 and 5 (P<0.05), and between groups 6 and 7 (P<0.05) being significant. Immunoblot analysis confirmed that the hepatic CYP1A2 level in group 6 was remarkably increased as compared to that in group 7. In addition, liver S9 from rats in group 6 consistently increased the mutagenic activities of six HCAs including MeIQx as compared to those in group 7. Thus, our results clearly indicate that CS enhances hepatocarcinogenesis when given in the initiation phase via increasing intensity of metabolic activation for MeIQx and possibly colon carcinogenesis when given in the post‐initiation phase in rats induced by MeIQx.
Mammary Carcinomas Induced in Human c‐Ha‐ras Proto‐oncogene Transgenic Rats Are Estrogen‐independent, but Responsive to d‐Limonene Treatment
Authors: Asamoto Makoto , Ota Tomonori , Toriyama‐Baba Hiroyasu , Hokaiwado Naomi , Naito Akihiro , Tsuda Hiroyuki
Published: August 2005

We have previously shown that transgenic rats carrying three copies of the human c‐Ha‐ras proto‐ oncogene (Hras128) are highly susceptible to N‐methyl‐N‐nitrosourea (MNU) mammary carcino‐ genesis. All transgenic rats treated with 50 mg/kgMNU, i.v. at 50 days of age, were found to rapidly develop multiple, large mammary carcinomas within as short a period as 8 weeks. In the present study, the effects of ovariectomy and treatment with d‐limonene, known to inhibit mammary carcinogenesis in non‐transgenic female rats, were investigated in Hras128 animals treated withMNU to clarify the role of the human c‐Ha‐ras proto‐oncogene and to characterize the induced mammary carcinomas. Although ovariectomy completely inhibited development of mammary carcinomas in their wild‐type counterparts, it did not affect either the incidence or the multiplicity of the mammary carcinomas in the Hras128 rats. On the other hand, treatment with d‐limonene, an inhibitor of ras protein isoprenylation, inhibited the breast tumor development. These results indicate that aberrant c‐Ha‐ras gene expression is involved in ovarian hormone‐independent growth and c‐Ha‐ras protein isoprenylation plays an important role in mammary carcinogenesis
Chemopreventive Effect of Lactobacttlus rhamnosus on Growth of a Subcutaneously Implanted Bladder Cancer Cell Line in the Mouse
Authors: Lim Boon‐Kian , Mahendran Ratha , Lee Yuan‐Kun , Bay Boon‐Huat
Published: August 2005

Lactic acid bacteria are known to have beneficial effects on the host, such as preventing carcinogenesis. The present study was designed to evaluate the chemopreventive effects of Lactobacttlus rhamnosus strain GG (LGG) in suppressing bladder cancer formation in a murine subcutaneous model of bladder cancer involving the inoculation of MB49 cells in C57B/L6 mice. After tumor implantation, one group of mice (n=8) was fed LGG immediately. The remaining mice that had tumors between 0.03–0.1 cm3 were divided into two groups: those fed LGG after 7 days (n=7) and those fed saline (n=7). A second group of mice without any inoculation of MB49 cells was fed either LGG (n=10) or saline (n=10) and served as non‐tumor‐bearing controls. LGG was administered orally at 1.6×l08 colony‐forming units daily. Mice fed LGG immediately after tumor cell implantation formed smaller tumors and some did not develop tumors (2 out of 8 mice), when the tumor burden was small. The level of spleen CD3, CD4 and CD8a T lymphocytes, as well as natural killer cells in mice fed immediately with LGG was also higher than that in control tumor‐bearing mice. There was an increase in lymphocytes and granulocytes in tumor sections, especially from the immediately fed group as compared to the controls. Our results suggest that oral consumption of LGG may prevent tumor growth via modulation of the immune system. The potential of LGG as an adjunct therapy in the treatment of bladder cancer could be further explored.
Chemopreventive Effects of a Flavonoid Antioxidant Silymarin on N‐Butyl‐N‐(4‐ hydroxybutyl)nitrosamine‐induced Urinary Bladder Carcinogenesis in Male ICR Mice
Authors: Vinh Pham Quang , Sugie Shigeyuki , Tanaka Takuji , Hara Akira , Yamada Yasuhiro , Katayama Masaki , Deguchi Takashi , Mori Hideki
Published: August 2005

The modifying effects of dietary administration of a flavonoid antioxidant, silymarin, a mixture of three flavonoids isolated from milk thistle seeds, on N‐butyl‐.N‐(4‐hydroxybutyl)nitrosamine (OH‐ BBN)‐induced urinary bladder carcinogenesis were examined in male ICR mice. Animals were divided into 5 groups, and groups 1 to 3 were given OH‐BBN (500 ppm) in drinking water for 6 weeks. Mice in group 2 were fed a diet containing 1000 ppm silymarin for 8 weeks during the initiation phase starting 1 week before OH‐BBN exposure, and mice in group 3 were fed the diet for 24 weeks during the postinitiation phase. Animals in group 4 were given only the test compound, and those in group 5 were given the basal diet alone throughout the experiment. Animals were sacrificed at the end of week 32. The frequency of bladder lesions, cell proliferation and cell cycle progression activity estimated in terms of the 5‐bromodeoxyuridine (BrdU) labeling index or cyclin Dl‐positive cell ratio were compared among the groups. Administration of silymarin in the initiation or postinitiation phase significantly decreased the incidences of bladder neoplasms and preneoplastic lesions. Dietary exposure to this agent significantly reduced the labeling index for BrdU and the cyclin Dl‐positive cell ratio in various bladder lesions. These findings suggest that silymarin is effective in preventing OH‐BBN‐induced bladder carcinogenesis in mice.