bluuebunny/crossref_metadata_2025 huggingface dataset:
Cytokeratin 19 expression in hepatocellular carcinoma predicts early postoperative recurrence
Authors: Uenishi Takahiro , Kubo Shoji , Yamamoto Takatsugu , Shuto Taichi , Ogawa Masao , Tanaka Hiromu , Tanaka Shogo , Kaneda Kenji , Hirohashi Kazuhiro
Published: August 2005

Clinicopathologic features and postoperative outcomes were investigated for patients who underwent curative surgery for biliary marker (CK7 and CK19)‐positive hepatocellular carcinoma (HCC). Of 157 HCCs, 93 were CK7(‐)CK19(+), 49 were CK7(+)‐CK19(‐), 1 was CK7(‐)CK19(+), and 14 were CK7(+)‐CK19(+). Semiquantitative analysis of expression levels demonstrated a significant correlation between CK7 and CK19 expression. Of various clinicopathologic parameters, tumor differentiation exhibited a significant correlation with CK7 and CK19 expression. All 15 patients with CK19‐positive HCC also had anti‐HBc. Log‐rank test revealed that CK7 expression, CK19 expression, high aspartate aminotransferase (AST) activity, low albumin concentration, portal invasion, intrahepatic metastasis, and severe fibrosis (cirrhosis) reduced the tumor‐free survival rate. Multivariate analysis demonstrated that CK19 expression, intrahepatic metastasis, and severe fibrosis were independent predictors of postoperative recurrence, while CK7 expression was not. Twelve of 15 patients with CK19‐positive HCC had tumor recurrence within 2 years after surgery, a significantly higher incidence of early recurrence than for CK19‐negative HCC. The incidence of extrahepatic disease, especially lymph node metastasis, was significantly higher for patients with CK19‐positive HCC. These findings indicate that CK19 expression is a predictor of early postoperative recurrence due to increased invasiveness.
Population‐based study of relationship between hospital surgical volume and 5‐year survival of stomach cancer patients in Osaka, Japan
Authors: Nomura Etsuko , Tsukuma Hideaki , Ajiki Wakiko , Oshima Akira
Published: August 2005

Despite the large number of surgical operations for stomach cancer in Japan, no study based on data from population‐based cancer registries has been conducted regarding the relationship between hospital surgical volume for stomach cancer and patients’ survival. Using data from the Osaka Cancer Registry (OCR), we performed survival analyses on 28,608 patients aged 35–79 years old who underwent surgery for stomach cancer, according to the extent of disease at diagnosis (localized: cancer is confined to the original organ; regional: cancer spreads to regional lymph nodes; adjacent: cancer infiltrates to adjacent tissue; distant: cancer metastasizes to distant organs). The study was divided into four periods; 1975–79, 1980–84, 1985–89 and 1990–94, according to year of diagnosis. Hospitals were stratified into four groups by total number of stomach cancer operations during each of the study periods, so that the numbers of operations in each group would be approximately equal. Cox's proportional hazards model was used to examine the relationships between hospital volume and 5‐year survival according to the extent of disease, after adjustment for age at diagnosis and sex. Positive relationships between hospital volume and 5‐year survival were seen in the localized and regional groups during the period 1975–79. These associations, however, decreased in later periods and disappeared in 1990–94 except for the very‐low‐volume hospitals. Stomach cancer patients treated in these hospitals remained at significantly higher risk of death than in the high‐volume hospitals. In the case of the distant group, there was no clear relationship between hospital volume and 5‐year survival during the study periods.
Antitumor effect of arsenic trioxide in murine xenograft model
Authors: Kito Mariko , Matsumoto Kenji , Wada Naoko , Sera Kouichiro , Futatsugawa Shoji , Naoe Tomoki , Nozawa Yoshinori , Akao Yukihiro
Published: August 2005

Arsenic trioxide, As2O3 (ATO), has been established to be an effective agent for treating acute promyelocytic leukemia, but its effect on solid tumors has not been fully explored. In the present study in a murine xenograft system, we found that ATO significantly inhibited tumor growth of the inoculated human hepatocellular carcinoma cell line HuH7 when administered either intravenously or intratumorally. Pathological examination revealed that ATO induced extensive cell death in the tumor. Some of the dead cells in intratumorally ATO‐treated mice showed characteristic features of apoptosis, such as nuclear condensation and fragmentation, and were TUNEL‐positive. The measurement of arsenic by using particle induced X‐ray emission revealed that arsenic was accumulated more in the tumor than in brain, kidney or liver after the intravenous injection of ATO, which is consistent with the hemorrhagic cell death observed in ATO‐treated tumor tissues. Thus, ATO appears to have potential for the treatment of solid tumors, as well as hematopoietic malignancies.
Extragonadal germ cell tumors in Japan
Authors: Ebi Hiromichi , Nakata Masanobu , Tahara Makoto , Igarashi Tadahiko , Kawada Kenji , Itoh Kuniaki , Ueda Ryuzo , Minami Hironobu
Published: August 2005

Extragonadal germ cell tumors (EGCT) represent only 2–5% of adult germ cell malignancies. Some publications from Asia have reported inferior treatment outcomes compared to data from an international study group. To ascertain whether this is generally the case, here we analyze treatment outcomes for 30 Japanese patients with EGCT. The medical records of 30 patients (25 non‐seminomas and 4 pure seminomas) treated from 1992 to 2002 were reviewed retrospectively. All patients with seminoma achieved long survival except one who died of chemotherapy‐related sepsis. Ten and 11 patients with EGCT presented with mediastinal and retroperitoneal primary sites, respectively. The 5‐year overall survival (OS) and progression‐free survival (PFS) for non‐seminoma was 71% and 42%, respectively. The 5‐year OS and PFS was 60% and 44%, respectively, for 10 patients with mediastinal nonseminoma, and 91% and 48%, respectively, for patients with retroperitoneal nonseminoma. Tumor marker values on day 7 were available for 19 patients. Among the 19 patients in whom AFP or β‐HCG were measured on day 7, the values had declined in 12 patients and were transiently elevated in 7 patients compared to pretreatment values. The transient elevations of tumor markers were significantly associated with poor OS (P=0.02) and PFS (P=0.008). The treatment outcome of Japanese patients with EGCT seemed to be comparable to that reported from international studies, suggesting no difference between ethnic groups. Transient tumor marker elevations on day 7 predict poor survival in EGCT patients and may be a useful parameter for identifying patients requiring more aggressive treatment.
Significance of nodal metastatic tumor characteristics in nodal metastasis and prognosis of patients with invasive ductal carcinoma of the breast
Authors: Hasebe Takahiro , Sasaki Satoshi , Imoto Shigeru , Ochiai Atsushi
Published: August 2005

There is no study evaluating the significance of nodal metastatic tumor characteristics in tumor progression of invasive ductal carcinomas (IDCs) of the breast. The purpose of this study was to investigate whether nodal metastatic tumor characteristics play an important role in the tumor progression of IDCs. The subjects of this study were 205 IDC patients with nodal metastases. Significant associations with increased numbers of nodal metastases, and patient outcomes were evaluated by multivariate analyses, in comparison with well‐known histological parameters. The numbers of lymph nodes with extra‐nodal invasion and with extra‐nodal blood vessel tumor emboli, the distance of extra‐nodal blood vessel tumor emboli from the nodes, and the nodal tumor dimensions significantly increased the number of nodal metastases in the multivariate analysis (P<0.001). Cox multivariate analyses showed that the parameters which significantly increased hazard rates (HRs) of disease‐free survival (DFS), distant organ metastasis (DOM) and overall survival were 6 or more mi‐totic figures of nodal metastatic tumors (P<0.05). Six or more lymph nodes with extra‐nodal invasion, and an extra‐nodal blood vessel tumor emboli dimension of >0.6 mm significantly increased the HRs of DFS and DOM in multivariate analyses (P<0.05). The present study demonstrated the important roles of nodal metastatic tumors in the tumor progression of IDCs. (Cancer Sci 2003; 94: 181–187)
Analysis of p53, K‐ras, c‐kit, and β‐catenin gene mutations in sinonasal NK/T cell lymphoma in northeast district of China
Authors: Hoshida Yoshihiko , Hongyo Tadashi , Jia Xinshan , He Yanjiao , Hasui Kazuhisa , Dong Zhiming , Luo Wen‐Juan , Ham Maria Francisca , Nomura Taisei , Aozasa Katsuyuki
Published: August 2005

Recently we reported the different frequenties of p53 and c‐kit gene mutations among sinonasal NK/T cell lymphoma (NKTCL) in Korea, north China (Beijing), and Japan, suggesting some racial, environmental, or life‐style differences as a possible cause of nasal tumorigenesis. In this study, gene mutations in p53, c‐kit, K‐ras, and β‐catenin gene were analyzed by polymerase chain reaction (PCR)‐single strand conformation polymorphism (SSCP) followed by direct sequencing in 20 cases of sinonasal NKTCL from northeast China (Shen Yang). Age of patients ranged from 5 to 63 (median, 40.0) years. p53 gene mutations were found in eight of 20 cases (40%), with exon 4 involvement in 10% of cases. The majority was missense mutations and G:C to A:T transition was predominant. The frequency of the c‐kit and K‐ras gene mutations was low (5%), while that of the β‐catenin gene was six of 20 cases (30%). From these findings, it is concluded that nasal NKTCL in northeast China shared common features with that in Korea in the younger onset of disease compared to that in Japan and lower frequency of p53 gene mutations with infrequent exon 4 involvement compared to that in Japan and north China. These differences might be caused by migration of susceptible populations or some environmental confounding factors. (Cancer Sci 2003; 94: 297–301)
Frequent occurrence of CCND1 deregulation in patients with early stages of plasma cell dyscrasia
Authors: Miura Kazuhisa , Lida Shinsuke , Hanamura Ichiro , Kato Miyuki , Banno Shogo , Ishida Takashi , Kusumoto Shigeru , Takeuchi Genji , Miwa Hiroshi , Nitta Masakazu , Inagaki Hiroshi , Eimoto Tadaaki , Nomura Kenichi , Taniwaki Masafumi , Ueda Ryuzo
Published: August 2005

Chromosomal translocations involving the immunoglobulin heavy chain gene (IgH) and nonrandom protooncogene loci are the hallmark of genetic alterations found not only in multiple myeloma (MM), but also in premalignant stages of MM, including monoclonal gammopathy of undetermined significance (MGUS) and smoldering myeloma (SMM). We studied the frequency of IgH (14q32) rearrangements and their partner chromosomes in 16 Japanese patients with MGUS (13 cases), and SMM (3 cases) by means of interphase double‐color fluorescence in situ hybridization (DCFISH) applied to purified plasma cells and using CD138‐bead selection. IgH rearrangement was recognized in nine of the patients (56.3%). Protooncogene loci juxtaposed to IgH were identified in seven cases including CCND1 (11q13) in six cases and FGFR3 (4p16) in one. Four out of the six t(11;14)‐positive cases showed nuclear staining of the cyclin D1 protein, whereas none of the seven t(11;14)‐negative cases did. Moreover, neither MUM1(6p25)‐IgH nor MAFB(20q11)‐IgH fusion signals were observed. This suggests to us that cyclin D1 deregulation due to the presence of t(11;14) is involved in the early development of plasma cell neoplasms, and that this event alone is not enough for the development of symptomatic myeloma. (Cancer Sci 2003; 94: 350‐354)
An efficient method to prepare T cell receptor gene‐transduced cytotoxic T lymphocytes type 1 applicable to tumor gene cell‐therapy
Authors: Tsuji Takemasa , Chamoto Kenji , Funamoto Hiromi , Kosaka Akemi , Matsuzaki Junko , Abe Hiroyuki , Fujio Keishi , Yamamoto Kazuhiko , Kitamura Toshio , Togashi Yuji , Koda Toshiaki , Nishimura Takashi
Published: August 2005

Genes encoding 2C T cell receptor (TCR) α, β chains from H‐2b‐re‐stricted Ld‐specific CD8+ cells were successfully transduced into polyclonally activated CD8+ cells by retroviral modification to generate antigen‐specific cytotoxic T lymphocytes (CTL). Antigen‐nonspecific CD8+ T cells polyclonally expanded in the presence of interleukin (IL)‐2, Th1 cytokines (interferon (IFN)‐γ and IL‐12) and anti‐IL‐4 monoclonal antibody showed neither cytokine production nor cytotoxicity in response to Ld‐expressing P815 tumor cells. However, 2C‐TCR gene‐modified CD8+ T cells exhibited both IFN‐γ production and cytotoxicity in response to P815 tumor cells. The antitumor activity of TCR gene‐modified Tc1 cells was also demonstrated in vivo by Winn's assay. Thus, we have developed an efficient method to induce TCR gene‐modified antigen‐specific Tc1 cells that exhibit antitumor activity both in vitro and in vivo. (Cancer Sci 2003; 94: 389–393)
New aspects of IFN‐α/β signalling in immunity, oncogenesis and bone metabolism
Authors: Takaoka Akinori , Taniguchi Tadatsugu
Published: August 2005

Although interferons (IFNs) were originally identified as humoral factors that confer an antiviral state upon cells, they have been demonstrated to be multifunctional in a variety of biological systems. The IFN‐α/β system modulates not only the cellular immune response to viral and bacterial infections, but also the oncogenic process and bone metabolism. Further studies have revealed additional unique facets of the IFN‐α/β system. A weak signal by constitutively produced IFN‐α/β is critical not only for the regulation of cellular amplification of IFN‐α/β production upon viral infection or the enhancement of signalling by other cytokines, but also for the regulation of adaptive immune responses, such as the enhancement of CD8+ T cell activation. Furthermore, IFN‐β signalling is critical for the regulation of the bone‐resorbing osteo‐clasts. In this review, we focus on the newly discovered roles of the IFN‐α/β system in host defense and bone remodeling, particularly on the functions of the weak IFN‐α/β signalling in the context of what we refer to as the “rewing‐up” model. (Cancer Sci 2003; 94: 405–411)
Expression of liver‐intestine cadherin and its possible interaction with galectin‐3 in ductal adenocarcinoma of the pancreas
Authors: Takamura Masaaki , Sakamoto Michlle , Ino Yoshinori , Shimamura Takeshi , Ichida Takafumi , Asakura Hitoshi , Hirohashi Setsuo
Published: August 2005

Liver‐intestine (LI) cadherin represents a novel type of cadherin within the cadherin superfamily, and is distinguished from other cadherins by specific structural and functional features. Among normal tissues, LI‐cadherin is known to be expressed in the intestinal mucosa, while its expression in cancerous tissue has not been investigated to date, except in gastric carcinoma. In the present study we investigated LI‐cadherin expression immunohis‐tochemically using our newly established monoclonal antibody in a large set (n=102) of tumor specimens from patients with ductal adenocarcinoma of the pancreas, and correlated the findings with the patients' survival. LI‐cadherin expression was seen focally in normal pancreatic ducts. In carcinoma, well‐differentiated carcinoma cases strongly expressed LI‐cadherin, whereas less differentiated areas and poorly differentiated carcinoma cases expressed less or were negative. Kaplan‐Meier analysis for all patients demonstrated that high Ll‐cadherin expression (>25% of cells stained positive) correlated with good survival (P<0.001). Cox regression analyses demonstrated that LI‐cadherin expression was one of the strongest predictors of outcome, independent of all other variables, and low Ll‐cadherin expression correlated with tumor de‐differentiation and advanced stage. Furthermore, galectin‐3 was identified as being coimmunoprecipitated with Ll‐cadherin and this interaction was inhibited by lactose in a dose‐dependent manner, but not by sucrose. Because galectin‐3 has been observed to show a similar expression pattern to LI‐cadherin in ductal adenocarcinoma of the pancreas, expression of LI‐cadherin and this interaction could have some role in ductal adenocarcinoma of the pancreas. (Cancer Sci 2003; 94: 425–430)
A report of two bronchioloalveolar carcinoma cases which were rapidly improved by treatment with the epidermal growth factor receptor tyrosine kinase inhibitor ZD1839 (“Iressa”1
Authors: Yano Seiji , Kanematsu Takanori , Miki Toyokazu , Aono Yoshinori , Azuma Masahiko , Yamamoto Akihiko , Uehara Hisanori , Sone Saburo
Published: August 2005

Bronchioloalveolar carcinoma (BAC), a form of pulmonary adeno‐carcinoma, presents unique clinical features, such as endobron‐chial spread and bronchorrhea in advanced stages. The prognosis for BAC patients in advanced stages is poor, as is the case for patients with other non‐small‐cell lung cancer (NSCLC) types, because of low susceptibility to conventional chemotherapy. Recently, an orally active, selective epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (EGFR‐TKI), ZD1839 (“Iressa”), has been investigated in phase II clinical studies (IDEAL 1 and IDEAL 2) as monotherapy against chemotherapy‐refractory NSCLC, and provided clinically significant antitumor activity. In this study, we examined the therapeutic efficiency of ZD1839 in chemotherapy‐refractory BAC patients with bronchorrhea. Two female BAC patients with bronchorrhea were treated once daily with ZD1839 (250 mg/day). In both cases, serous sputum production was dramatically reduced within 3 days of starting the treatment, and hypoxia and radiographic signs of bilateral lung consolidation were visibly improved within 7 days. Following more than 8 months of treatment, no evidence of recurrence or severe adverse events has been observed. These results suggest that this selective EGFR‐TKI, ZD1839, may be a powerful agent for treatment of chemotherapy‐refractory BAC patients with bronchorrhea. (Cancer Sci 2003; 94: 453–458)
Overexpression of the Wilms' tumor gene WT1 in primary thyroid cancer
Authors: Oji Yusuke , Miyoshi Yasuo , Koga Satoko , Nakano Yoko , Ando Akiko , Nakatsuka Shin‐ichi , Ikeba Ai , Takahashi Eigo , Sakaguchi Nao , Yokota Asumi , Hosen Naoki , Ikegame Kazuhiro , Kawakami Manabu , Tsuboi Akihiro , Oka Yoshihiro , Ogawa Hiroyasu , Aozasa Katsuyuki , Noguchi Shinzaburo , Sugiyama Haruo
Published: August 2005

The expression levels of the Wilms' tumor gene WT1 were examined in 34 primary thyroid cancers (24 papillary, 5 follicular, 1 anaplastic, and 4 medullary carcinomas), 17 thyroid follicular adenomas, and 6 normal‐appearing thyroid tissues using quantitative real‐time reverse transcriptase‐polymerase chain reaction (RT‐PCR). In 33 of 34 thyroid cancers, the WT1 mRNA was expressed at levels ranging from 5.0×10−5 to 8.3×10−2 levels (WT1 expression level in K562 leukemic cells was defined as 1.0). The WT1 mRNA expression levels were significantly higher than those in either thyroid follicular adenomas (P<0.001) or normal‐appearing thyroid tissues (P<0.01). Immunohistochemical analysis confirmed the expression of WT1 protein in 20 of 21 thyroid cancers with WT1 mRNA expression. WT1 protein was also detected in 6 of 7 follicular adenomas with WT1 mRNA expression. However, the intensity of staining of WT1 protein in adenoma cells was weaker than that in cancer cells and its expression was restricted to approximately 30–80% of adenoma cells in the tumors examined. The direct sequencing analysis of the WT1 genomic DNA showed no mutations in any of the 10 exons of the WT1 gene in all of the 9 different thyroid cancers. These findings indicate an important role of the wild‐type WT1 gene in the tumorigenesis of primary thyroid cancer.
Reduction of CTLL‐2 cytotoxicity by induction of apoptosis with a Fas‐estrogen receptor chimera
Authors: Kametaka Minako , Kume Akihiro , Okada Takashi , Mizukami Hiroaki , Hanazono Yutaka , Ozawa Keiya
Published: August 2005

Allogeneic bone marrow transplantation and donor lymphocyte infusion are powerful treatments for chemotherapy‐resistant leukemia. Tumor eradication is attributed to a graft‐versus‐leukemia reaction by the donor‐derived cytotoxic T lymphocytes (CTLs), but the same cell population may cause severe graft‐versus‐host disease. One strategy to suppress harmful CTL activity is to incorporate a suicide gene into the donor lymphocytes prior to infusion, and to destroy these cells when they aggressively attack nonmalignant host tissues. In this study, we investigated the feasibility of using a Fas‐estrogen receptor fusion protein (MfasER) to control T cell‐mediated cytotoxicity, based on our previous finding that the chimera transmits a Fas‐mediated death signal through activation by estrogen binding. A murine CTL line CTLL‐2 was transfected with a vector encoding MfasER, and the growth, viability and cytotoxic activity of the transfected cells (CTLL/MfasER) were analyzed. The expression of apoptosis‐related proteins such as Fas ligand and perforin was also investigated. In the absence of estrogen, CTLL/MfasER showed similar growth to parental CTLL‐2, and the killing activity was preserved. Addition of 10−7M estrogen induced a rapid apoptosis of CTLL/MfasER, and the cytotoxicity was severely impaired. A decrease of Fas ligand and perforin in the estrogen‐treated CTLL/MfasER was seen in an immunoblot analysis. These functional and biochemical analyses showed that the estrogen‐inducible apoptosis in MfasER‐expressing CTLs rapidly terminated their target cell killing. The feasibility of using the MfasER‐estrogen system to control graft‐versus‐host disease was demonstrated.
Overexpression of the Wilms' tumor gene WT1 in colorectal adenocarcinoma
Authors: Oji Yusuke , Yamamoto Hirofumi , Nomura Masaya , Nakano Yoko , Ikeba Ai , Nakatsuka Shin‐ichi , Abeno Sakie , Kiyotoh Eiji , Jomgeow Tanyarat , Sekimoto Mitsugu , Nezu Riichiro , Yoshikawa Yukinobu , Inoue Yoshifumi , Hosen Naoki , Kawakami Manabu , Tsuboi Akihiro , Oka Yoshihiro , Ogawa Hiroyasu , Souda Shigeo , Aozasa Katsuyuki , Monden Morito , Sugiyama Haruo
Published: August 2005

Expression of the Wilms' tumor gene WT1 was examined in 59 cases of colorectal adenocarcinoma to examine the involvement of WT1 in tumorigenesis. Quantitative real‐time reverse transcription‐polymerase chain reaction (RT‐PCR) showed that WT1 mRNA was expressed in the range from 7.2x10−5 to 4.9x10−1 levels (WT1 expression level in K562 leukemic cells was defined as 1.0) in all (100%) of the 28 cases of colorectal adenocarcinoma examined, and that the WT1 mRNA expression levels were higher in 20 (71%) of the 28 cases compared to those of normal‐appearing mucosal tissues examined. Immunohistochemical analysis using an anti‐WT1 antibody was performed on 46 cases of colorectal adenocarcinoma (15 of the 28 cases with WT1 mRNA expression and 31 newly collected cases), and the expression of WT1 protein was detected in 41 (89%) of the 46 cases. The direct sequencing analysis of the WT1 genomic DNA showed no mutations in any of the 10 exons of the WT1 gene in any of 5 different colorectal ade‐nocarcinomas. These results may indicate an important role of the wild‐type WT1 gene in tumorigenesis of colorectal adenocarcinoma.
A naturally occurring p73 mutation in a p73‐p53 double‐mutant lung cancer cell line encodes p73α protein with a dominant‐negative function
Authors: Huqun  , Endo Yoshiyuki , Xin Hong , Takahashi Mitsu , Nukiwa Toshihiro , Hagiwara Koichi
Published: August 2005

p73, a close homolog of p53 tumor suppressor, induces growth arrest and apoptosis. However, its role in cancers is controversial because of the rarity of p73 mutations, lack of tumors in p73‐knockout mice, and the presence of multiple isotypes, among which ΔN isotypes inhibit the function of TA isotypes. We analyzed three naturally occurring p73 mutants found in lung cancer cell lines, NCI‐H1155, DMS 92 and A427. NCI‐H1155 is a cell line that has a p73 mutation [p73(G264W)] in the DNA‐binding domain, as well as a p53 mutation [p53(R273H)], which is frequently found in human cancers and has a “gain‐of‐function” characteristic. p73α(G264W) not only lacks transactivation activity itself, but also suppressed the transactivation activity of the wild‐type p73α in a dose‐dependent manner, indicating that p73α(G264W) is a dominant‐negative mutant. p73α(G264W) failed to suppress colony formation. We tested two other mutations, p73(Del418) in DMS 92 and p73(Del603) in A427. Both mutants retained similar levels of transactivation activity and suppression of colony formation to those of wild‐type p73. The biological significance of these two mutations is unclear. In NCI‐H1155 cells the coexistence of mutations that abrogate the normal functions of p73 and p53 may indicate that each mutation confers an additive growth advantage upon the cells.
Comparison of gene expression profiles of lymphoma cell lines from transformed follicular lymphoma, Burkitt's lymphoma and de novo diffuse large B‐cell lymphoma
Authors: Maesako Yoshitomo , Uchiyama Takashi , Ohno Hitoshi
Published: August 2005

To determine the specific gene expression in B‐cell lymphoma subtypes, we compared expression profiles of cell lines from transformed follicular lymphoma (tFL), Epstein‐Barr virus‐negative (EBV(‐)) Burkitt's lymphoma (BL) and EBV(+)BL. Complementary DNAs were synthesized from these cell lines and hybridized with the Atlas Human 1.2 Array membrane. Hierarchical clustering analysis based upon the levels of 43 genes highlighted characteristic expression patterns of the 3 lymphoma subtypes. Genes expressed at higher levels in tFL than EBV(‐)BL and EBV(+)BL included calcium/calmodulin‐dependent protein kinase I (CAMK1) and mitogen‐activated protein kinase 10 (MAPK10). EBV(‐)BL was characterized by high‐level expression of amyloid β precursor protein (APP), heat shock 27 kD protein 1 (HSPB1) and mothers against decapentaplegic homolog 1 (MADH1). Gardner‐Rasheed feline sarcoma viral oncogene homolog (FGR) was the most significant gene to delineate EBV(+)BL. A subtype prediction algorithm using 34 genes correctly classified 22 (92%) of 24 lymphomas into FL/tFL, EBV(‐)BL or EBV(+)BL. By comparison with normal reference B‐cell materials, the expression patterns of the selected genes were characteristic of lymphomas. We extended the clustering analysis to cell lines from de novo diffuse large B‐cell lymphoma (DLBCL). The DLBCL cell lines were either separated from the former 3 lymphoma subtypes or segregated with EBV(+)BL, possibly reflecting variable genetic abnormalities. The associations of CAMK1 with tFL, APP and MADH1 with EBV(‐)BL, FGR with EBV(+)BL, and BCL2 with tFL and DLBCL were confirmed by real‐time quantitative reverse transcriptase‐medizted polymerase chain reaction assays. This study has provided new molecular markers, expressions of which are closely associated with B‐cell lymphoma subtypes.
Selective hyperthermia using magnetoliposomes to target cervical lymph node metastasis in a rabbit tongue tumor model
Authors: Hamaguchi Shigeaki , Tohnai Iwai , Ito Akira , Mitsudo Kenji , Shigetomi Toshio , Ito Masafumi , Honda Hiroyuki , Kobayashi Takeshi , Ueda Minoru
Published: August 2005

The effect of hyperthermia on cervical lymph node metastasis of VX7 tongue cancer in female Japanese white rabbits was investigated. Magnetoliposomes (MLs) with a neutral surface charge and a size of 94.1 nm were used as heating mediators. MLs were injected into the tongue 20 days after tumor transplantation, and we examined whether they reached the metastatic deep cervical lymph node. The highest magnetite concentration 24 h after ML injection was detected in the lymph node, followed by tongue, spleen, blood, and liver. Rabbits were separated into three groups: group I as the control; group II with ML injection alone; and group III with ML injection and hyperthermia 24 h after ML injection, generated by applying an alternating magnetic field (118 kHz, 384 Oe) to the neck region. The hyperthermic effect was evaluated in terms of the percentage of necrosis in proportion to the metastatic tumor and the apoptotic index (AI), defined as the ratio of TUNEL‐positive cells. The temperature of lymph nodes in group III reached over 44°C. The mean area of necrosis in group III was 58.0%, which was significantly higher than that in group I (19.6%) or group II (20.4%). The AI in group III was 22.9%, significantly higher than in group I (1.67%) or II (1.42%). The difference between group I and II was not statistically significant. Group III tumor sites around MLs showed necrosis or apoptosis‐positive cells induced by hyperthermia. These results indicate that MLs injected into the tongue can target cervical lymph node metastases and accumulate there at concentrations sufficient to generate therapeutically effective temperatures.
Cloning of Mongolian gerbil cDNAs encoding inflammatory proteins, and their expression in glandular stomach during H. pylori infection
Authors: Matsubara Satoshi , Shibata Hideyuki , Takahashi Mami , Ishikawa Fumiyasu , Yokokura Teruo , Sugimura Takashi , Wakabayashi Keiji
Published: August 2005

Mongolian gerbils are considered to be a good animal model for understanding the development of Helicobacter pylori‐associated diseases. However, limitations regarding the genetic information available for this animal species hamper the elucidation of underlying mechanisms. Thus, we have focused on identifying the nu‐cleotide sequences of cDNAs encoding Mongolian gerbil inflammatory proteins, such as interleukin‐1 (IL‐lβ), tumor necrosis factor a (TNF‐α), cyclooxygenase‐2 (COX‐2) and inducible nitric oxide synthase (iNOS). Furthermore, we examined the mRNA expression of these genes in the glandular stomach by RT‐PCR at 1–8 weeks after H. pylori infection. The deduced amino acid homol‐ogies to mouse, rat and human proteins were 86.2%, 83.6% and 67.8% for IL‐1β, 87.2%, 85.1% and 78.4% for TNF‐α, 91.9%, 90.2% and 84.8% for COX‐2 and 90.8%, 89.1% and 80.1% for iNOS, respectively. The average stomach weight of Mongolian gerbils inoculated with H. pylori was increased in a time‐dependent manner at 1, 2, 4 and 8 weeks after inoculation. In the py‐loric region, mRNA expression levels of IL‐1β, TNF‐α and iNOS were increased in H. pylori‐infected animals at the 2 weeks time point, while in the fundic region, expression levels of IL‐1β, TNF‐α and iNOS were elevated at 4 and 8 weeks. The COX‐2 expression level in the fundic region was clearly elevated in infected animals compared with control animals at 4 and 8 weeks, but in the py‐loric region, expression levels were similar in both infected and control animals. Thus, our results indicate that oxidative stress occurs from an early stage of H. pylori infection in the glandular stomach of Mongolian gerbils.
Eradication of Helicobacter pylori induces apoptosis and inhibits proliferation of heterotopic proliferative glands in infected Mongolian gerbils
Authors: Cao Xueyuan , Tsukamoto Tetsuya , Nozaki Koji , Shimizu Nobuyuki , Mizoshita Tsutomu , Kumagai Toshiko , Kaminishi Michio , Tatematsu Masae
Published: August 2005

Mongolian gerbils infected with Helicobacter pylori (H. pylori) develop heterotopic proliferative glands (HPGs) in the glandular stomach submucosa. To investigate the effects of H. pylori eradication on cell turnover in HPGs, three antibiotics, lansoprazole, amoxicillin and clarithromycin, were administered at 50 or 75 weeks after inoculation of H. pylori, and the stomachs were excised for histological examination at 1, 2, 4, 8 or 25 weeks thereafter. The HPGs were classified into gastric type (G‐type) and others (Gl+I‐type), which included both pure intestinal (I‐type) and gastric‐and‐intestinal mixed type (Gl‐type). Apoptosis and cell proliferation were evaluated by means of TUNEL assay and BrdU labeling, respectively. At 8 weeks post‐eradication, apoptotic indices were significantly increased in the eradication group (G‐type: 2.5%; Gl+I‐type: 7.2%) compared to the non‐eradication group (G‐type: 0.6%; Gl+I‐type: 2.1%: P<0.01), while BrdU labeling indices were significantly decreased (G‐type: 1.9%; Gl+I‐type: 6.8% as compared with 4.3% and 13.2%, respectively, P<0.01 for both). At 25 weeks, the apoptotic indices were similarly higher [G‐type: 0.4 (eradication group) vs. 0.2% (non‐eradication group); Gl+I‐type: 5.8 vs. 1.1%, both P<0.01], and the BrdU labeling indices (G‐type: 0.8 vs. 2.2%, P<0.01; Gl+I‐type: 5.1 vs. 11%, P<0.05) continued to be lower in HPGs. Furthermore, there were highly significant reductions in the areas of HPGs at 8 and 25 weeks post‐eradication. These findings demonstrated that eradication results in apoptosis and reduction of proliferation of HPGs in H. pylori‐infected gerbils, these lesions thus being apparently reversible through regulation of cell kinetics.
Influence of hospital procedure volume on ovarian cancer survival in Japan, a country with low incidence of ovarian cancer
Authors: Ioka Akiko , Tsukuma Hideaki , Ajiki Wakiko , Oshima Akira
Published: August 2005

The survival of ovarian cancer patients has been reported to be superior at hospitals with a high volume of operations. A population‐based study was carried out to assess whether this is true in Japan, where the incidence rate is relatively low as compared with other developed countries. The Osaka Cancer Registry's data were used to investigate associations between hospital procedure volume and survival of ovarian cancer patients. Hospitals were ranked according to the number of operations for ovarian cancer performed per year (high/medium/low/very low). Survival analysis was restricted to the reported 2450 cases who lived in Osaka Prefecture (except for Osaka City) diagnosed in 1975–1995, or those who resided in Osaka City in 1993–1995, since active follow‐up data on vital status 5 years after the diagnosis were available. The relative 5‐year survival for all ovarian cancer cases was 38.8%, and the survival was higher with greater hospital volume (22.3%/34.2%/46.2%/55.0%). After adjustment for age, histologic type and cancer stage by the Cox regression model, patients receiving care in very‐low‐volume hospitals were seen to have a 60% higher risk of death than patients receiving care in high‐volume hospitals (P<0.01). Although some limitations existed in this study, the results indicated that further centralization of operative treatment in high‐volume hospitals might improve survival of ovarian cancer patients in Japan.
Dephosphorylation of Bcl‐2 by protein phosphatase 2A results in apoptosis resistance
Authors: Simizu Siro , Tamura Yuki , Osada Hiroyuki
Published: August 2005

The anti‐apoptotic protein, Bcl‐2 was phosphorylated at the Ser‐87 residue in normal human blood cells, while it was not phosphorylated in tumor cells. We identified protein phosphatase 2A (PP2A) as a Bcl‐2‐associated phosphatase that is responsible for dephosphorylation of Bcl‐2 in tumor cell lines. Treatment of the tumor cells with a PP2A inhibitor resulted in the appearance of Bcl‐2 phosphorylation at Ser‐87. This observation suggests that Bcl‐2 is constitutively phosphorylated, but is immediately dephosphorylated by PP2A in tumors. Phosphorylation of Bcl‐2 protein at the Ser‐87 residue resulted in a reduction in anti‐apoptotic function in human tumor cell lines. Thus, not only the expression level, but also the dephosphorylation status may have important implications for the oncogenic activity of Bcl‐2.
Isolation of p53‐target genes and their functional analysis
Authors: Nakamura Yusuke
Published: August 2005

Mutations of the p53 gene are the most common genetic alterations found in human cancers, and are known to play crucial roles in tumor development and progression. The p53 gene encodes a protein functioning as a transcription factor, and the biological functions of p53 are manifested through the activities of its downstream genes. Identification of these downstream genes involved in the p53‐signaling pathway should provide more detailed insight into the molecular mechanisms that mediate tumor‐suppressor activities, as well as various responses to cellular stress. We have been attempting to isolate p53‐target genes by means of various approaches, including differential display, cDNA microarray analysis, and direct cloning of the p53‐binding sequences from human genomic DNA. Here I review our recent work on isolation of p53‐target genes and their functional analysis. The physiological functions of p53‐target genes include apoptosis (GML, p53AIP1, and STAG1), DNA repair (p53R2), inhibition of angiogenesis (BAI1), re‐entry into the cell cycle (p53RFP), oxidative stress (CSR), and determination of cell fate (p53RDL1). (Cancer Sci 2004; 95: 7–11)
Nemo‐like kinase suppresses a wide range of transcription factors, including nuclear factor‐kB
Authors: Yasuda Jun , Yokoo Hideki , Yamada Tesshi , Kitabayashi Issay , Sekiya Takao , Ichikawa Hitoshi
Published: August 2005

Nemo‐like kinase (NLK) is a serine/threonine kinase that suppresses the transcription activity of the β‐catenin‐T‐cell factor (TCF) complex through phosphorylation of TCF. Our previous study showed that NLK overexpression induces apoptosis in DLD‐1 human colon cancer cells and that apoptosis induction presumably requires a mechanism other than the suppression of β‐catenin‐TCF complex. Luciferase reporter gene assay with pNF‐kB‐Luc revealed that NLK could suppress transcription activity of NF‐kB in a kinase‐dependent manner. However, it appeared that transcription co‐activators of NF‐kB, such as CREB binding protein (CBP)/p300, were likely to be the direct targets of NLK, rather than NF‐kB itself. Luciferase reporter gene analysis of GAL4‐CBP fusion proteins revealed that the C‐terminal region of CBP was critical for transcription suppression by NLK. In vitro kinase assay showed that NLK could phosphorylate the C‐terminal domain of CBP. However, HAT activity was not suppressed by the induction of wild‐type NLK in DLD‐1 cells. Furthermore, we observed that NLK suppressed the transcription activity of AP‐1, Smad, and p53, all of which also utilize CBP as a co‐activator. The extent of suppression by NLK was similar among the transcription factors tested (50–60% reduction). Our results suggest that NLK may suppress a wide range of gene expression, possibly through CBP. (Cancer Sci 2004; 95: 52–57)
Cloning of a G‐protein‐coupled receptor that shows an activity to transform NIH3T3 cells and is expressed in gastric cancer cells
Authors: Okumura Shun‐ichiro , Baba Hiroko , Kumada Tatsuro , Nanmoku Koji , Nakajima Hirofumi , Nakane Yasushi , Hioki Koshiro , Ikenaka Kazuhiro
Published: August 2005

The present study was directed towards the identification of novel factors involved in the transformation process leading to the formation of gastric cancer. A cDNA library from human gastric cancer cells was constructed using a retroviral vector. Functional cloning was performed by screening for transformation activity in transduced NIH3T3 cells. Six cDNA clones were isolated, including one encoding the elongation factor 1asubunit, which was already known to play a role in tumorigenesis. One cDNA (clone 56.2), which was repeatedly isolated during the course of screening, encoded a protein identical to a G‐protein‐coupled receptor protein, GPR35. In addition, another cDNA clone (72.3) was found to be an alternatively spliced product of the GPR35 gene, whereby 31 amino acids were added to the N‐terminus of GPR35. Hence, the proteins encoded by clones 56.2 and 72.3 were designated GPR35a and GPR35b, respectively. RT‐PCR experiments revealed that GPR35 gene expression is low or absent in surrounding non‐cancerous regions, while both mRNAs were present in all of the gastric cancers examined. The level of 72.3‐encoded mRNA was consistently significantly higher than that of 56.2 encoded mRNA. An expression pattern similar to that observed in gastric cancers was detected in normal intestinal mucosa. Based on the apparent transformation activities of the two GPR35 clones in NIH3T3 cells, and the marked up‐regulation of their expression levels in cancer tissues, it is speculated that these two novel isoforms of GPR35 are involved in the course of gastric cancer formation.
Treatment efficiency of a suicide gene therapy using prostate‐specific membrane antigen promoter/ enhancer in a castrated mouse model of prostate cancer
Authors: Ikegami Shusei , Tadakuma Takushi , Ono Takeshi , Suzuki Satoshi , Yoshimura Ichiro , Asano Tomohiko , Hayakawa Masamichi
Published: August 2005

Suicide gene therapy has potential for the treatment of prostate cancer under conditions of androgen deprivation. We show here that the combination of promoter/enhancer of prostate‐specific membrane antigen (PEPM) and the Cre‐loxP system is a good method to express a suicide gene, namely herpes virus thymidine kinase (TK), in prostate cancer cells. We have examined this system in a castration model in vivo, in comparison with a prostate‐specific antigen promoter/enhancer system (PP). In the castrated mice, the tumor luciferase activity with the combination of the PEPM plus the Cre‐loxP system was about 50 times greater than that with the control GL3 plasmid. A similar increase was observed in non‐castrated mice. In contrast, the luciferase activity of the plasmid PP was decreased significantly in tumors from castrated mice as compared with tumors from non‐castrated control mice. Regarding the therapeutic effect, the combination plasmid PEPM‐Cre plus CMV‐loxP‐TK exhibited a strong inhibitory effect on tumor growth in the castrated mice, as in the non‐castrated mice. In contrast, PP‐TK plasmid did not show any significant growth inhibition in the castrated mice. These findings indicate that the combination of PEPM and Cre‐loxP system may have a good treatment effect under androgen ablation conditions in vivo, and our system may therefore be applicable to patients who have previously received androgen deprivation therapy.
Estrogen enhancement of androgen‐responsive gene expression in hormone‐induced hyperplasia in the ventral prostate of F344 rats
Authors: Fujimoto Nariaki , Suzuki Tomoharu , Honda Hiroaki , Kitamura Shigeyuki
Published: August 2005

It has been postulated that, in addition to the crucial role of androgens, estrogens may be involved in development of prostate hyperplasia and cancer. In rats, combined administration of estrogen and androgen synergistically increases ventral prostate weight, and continued treatment results in the development of glandular hyperplasia. Prostate adenocarcinoma can be induced by chemical carcinogens in rats, and estrogen given together with an androgen generally shortens the latent period or increases the incidence and/or multiplicity of carcinomas. However, the mechanisms responsible for these synergistic effects of estrogen and androgen are poorly understood. In the present study, we examined the combined effects of 17β‐estradiol (E2) and testosterone (T) on gene expression in an early stage of prostate hyperplasia in an F344 rat model. ERα expression, which has been suggested to contribute to development of prostatic hyperplasia, was increased by the combined treatment with T and E2, while it was suppressed by T alone. Expression levels of two androgen‐responsive genes, probasin and kallikrein S3, were increased in the ventral prostate of rats treated with T plus E2 for 4 weeks in a dose‐dependent manner, while short‐term treatment did not alter the expression. These results suggested that enhancing effects of E2 on transcription of androgen‐responsive genes, as well as an increased level of ERα may play roles in the synergistic effects of E2 on T‐induced prostate hyperplasia.
Sonodynamic therapy on chemically induced mammary tumor: Pharmacokinetics, tissue distribution and sonodynamically induced antitumor effect of porfimer sodium
Authors: Yumita Nagahiko , Okuyama Nobuo , Sasaki Kazuaki , Umemura Shin‐ichiro
Published: August 2005

The sonodynamically induced antitumor effect of porfimer sodium (PF) was evaluated on a chemically induced mammary tumor in Sprague‐Dawley rats. The timing of 24 h after the administration of PF was chosen for the ultrasonic exposure, based on pharmacokinetic analysis of the PF concentrations in the tumor, plasma, skin and muscle. At a PF dose not less than 2.5 mg/kg and at a free‐field ultrasonic intensity not less than 3 W/cm2, the synergistic effect between PF administration and ultrasonic exposure on the tumor growth inhibition was significant. The ultrasonic intensity showed a relatively sharp threshold for the synergistic antitumor effect, which is typical of an ultrasonic effect mediated by acoustic cavitation. These results suggest that PF is a potentially useful as a sonosensitizer for sonodynamic treatment of chemically induced tumors.
The effect of cimetidine or omeprazole on the pharmacokinetics of escitalopram in healthy subjects
Authors: Malling D. , Poulsen M. N. , Søgaard B.
Published: August 2005

Aims

To investigate the effects of co‐administration of cimetidine or omeprazole on the pharmacokinetics of escitalopram.

Methods

Two randomized placebo‐controlled crossover studies were carried out. Sixteen healthy subjects were administered placebo, or cimetidine (400 mg twice daily) for 5 days (study 1) or omeprazole (30 mg once daily) for 6 days (study 2). On day 4 (study 1) or day 5 (study 2), a single dose of escitalopram (20 mg) was administered. Blood samples were taken at predetermined times for the measurement of serum concentrations of escitalopram and its demethylated metabolite (S‐DCT). Treatment‐emergent adverse events were also monitored.

Results

Co‐administration with cimetidine caused a moderate increase in the systemic exposure [AUC(0, ∞)] to escitalopram (geometric mean ratio = 1.72, [95% CI 1.34, 2.21]) and a small increase in t½ from 23.7 to 29.0 h (5.24 h [3.75, 6.70]). Co‐administration with omeprazole also resulted in a moderate increase in the escitalopram AUC(0, ∞) (1.51 [1.39, 1.64]) and a small increase in t½ from 26.5 to 34.8 h (8.3 h [6.44, 10.2]). There was no significant change in S‐DCT AUC(0, ∞) after co‐administration of either cimetidine or omeprazole. Co‐administration of cimetidine or omeprazole had no effect on the incidence of treatment‐emergent adverse events.

Conclusions

In view of the good tolerability of escitalopram, the pharmacokinetic changes observed on co‐administration with cimetidine or omeprazole are unlikely to be of clinical concern.
Dwarf and giant geckos from the cellular perspective: the bigger the animal, the bigger its erythrocytes?
Authors: STAROSTOVÁ ZUZANA , KRATOCHVÍL LUKÁŠ , FRYNTA DANIEL
Published: August 2005

Although evolutionary and ecological consequences of body size changes are relatively well understood, the proximate mechanisms of body size alteration at a cellular level are often surprisingly neglected, especially in vertebrates. The question of whether larger animals are made from larger cells is rarely tested in an explicit phylogenetic framework, i.e. among closely related species with known phylogeny.

Here we explore the relationship between erythrocyte and body size in a small gecko family (Eublepharidae) exhibiting large body size variation.

We found positive interspecific correlation of cell and body size. Assuming that size of other cell types changed in a similar way to red blood cell size, we can conclude thanc.15–20% of body size change in this group could be attributed to cell size variation.

As larger cells are generally more frugal than smaller cells, we hypothesise that a macro‐evolutionary trade‐off exists between body complexity and energetic efficiency.

We believe that knowing how particular animal lineages have solved this trade‐off during body size evolution will help us explain much of the variation in ecophysiological traits among clades as well as within them.