Biomedical subjects

Tetsuya Toge

Publications and source records attributed to Tetsuya Toge.

84 records · Page 5Linked to original sources

PubMedDate not supplied

Quantitative detection of carcinoembryonic antigen messenger RNA in the peritoneal cavity of gastric cancer patients by real-time quantitative reverse transcription polymerase chain reaction.

BACKGROUND: Peritoneal dissemination is the most frequent cause of recurrence in gastric cancer patients after gastrectomy. We have previously demonstrated the prognostic factors in gastric cancer including growth factors and their receptors and intra-peritoneal CEA (1,2). MATERIALS AND METHODS: In the present study, to elucidate whether CEA mRNA in peritoneal lavage is a good marker for the prediction of peritoneal recurrence of gastric cancer patients, quantitative real-time RT-PCR with primers specific for CEA was performed by peritoneal lavage of 124 gastric cancer patients collected at the time of operation. RESULTS: CEA crossing-point values (CEA-CP) were significantly correlated with the depth of tumor invasion and the values were cut-off at 27.11, which was the minimum value of mucosal cancer patients. The CEA-CP was significantly correlated with lymph node metastasis, lymphatic invasion, venous invasion, stages and overall survivals. In prediction of peritoneal recurrence by CEA-CP, sensitivity and specificity were 76.9% and 67.7%, respectively. Cytology predicted peritoneal recurrence with 23.1% sensitivity and 98.1% specificity. CONCLUSION: Quantitative RT-PCR of CEA mRNA of peritoneal lavage is a useful factor for prognosis and predicting peritoneal recurrence.

Biomarkers, Tumor↗

PubMedDate not supplied

Clinical relevance of meta-analysis of survival benefit when deciding treatment of adjuvant chemotherapy in gastric cancer.

The results of meta-analysis regarding survival benefit in adjuvant chemotherapy for gastric cancer are very useful and informative in clinical practice. However, in considering the current status of adjuvant therapy the following items regarding the effects of adjuvant therapy need to be considered: (i) the assessment of survival effect to adjuvant chemotherapy might differ due to the different lymph node dissection protocols practiced; (ii) the actual effect of adjuvant therapy on residual cancer cells cannot be monitored during treatment without evaluating long-term survival, although the effect of adjuvant therapy is dependent on the amount of residual cancer cells and their drug-sensitivity. The development of new regimens that include molecular targeted drugs may help to demonstrate a survival benefit of adjuvant chemotherapy. The studies collected for a meta-analysis are heterogeneous in their design and should not be thought of as providing estimates of a single quality. Since actual potential survival benefit of adjuvant chemotherapy most likely does exist, a large randomized controlled trial using new rationalized regimens based on standardized lymph node dissection is required for the establishment of survival benefit of adjuvant therapy in patients with advanced gastric cancer.

Clinical Trials as Topic↗

PubMedDate not supplied

Factors that influence the incidence of microscopic carcinoma in frozen and preserved specimens from patients with breast carcinoma after breast-conserving surgery.

Because a positive margin is an important risk factor for decreased survival of patients with breast cancer after breast-conserving surgery (BCS), we examined the rate of positive surgical margins after BCS, determined from intraoperative frozen and preserved sections, and we assessed factors that influence the incidence of positive margins in surgical specimens. The cases of 172 patients who underwent BCS between June 1989 and March 2001 were studied. The pathologic features and positive margins were evaluated by microscopic examination of frozen and preserved sections. The grade of each positive margin was arbitrarily classified into one of four groups: i) cancer involved; ii) closed margin (<2 mm); iii) partially closed margin (>3 mm and <5 mm) and iv) free margin (>5 mm). Biomarkers were estimated by immunohistochemistry and DNA ploidy, and the relationship of clinical parameters was statistically analyzed using the chi(2) test. In 59 of 172 cases (34.3%), the surgical margin was positive: horizontal in 47 cases (27.3%) and vertical in 12 cases (6.9%). The free margin was significantly correlated with positive status of estrogen and progesterone receptors and middle age (35-50 years) in the patients, while clinicopathologic factors were not correlated with positive margins in surgical specimens. Although 17 cases of cancer were found in 9 frozen specimens and 8 preserved sections, the mammographic findings showed spiculated masses in 7 of the 9 frozen specimens (77.7%) and 4 of the 8 preserved sections (50%), and microcalcification in 2 frozen specimens (22.2%) and 4 preserved sections (50%). The overexpression (2+ or 3+) of the HER-2 protein was found in 8 of the 9 frozen specimens (87.5%) and in 7 of the 8 preserved sections (88.8%). In the 11 cases with spiculated masses, extensive intraductal components (EICs) of various grades were found; the presence of EICs in a spiculated lesion was observed in 5 of the 7 frozen specimens and in all 4 of the preserved sections. The local extension of breast carcinoma cells in BCS is associated in part with the findings of spiculation and microcalcification, which are associated with EICs. The overexpression of HER-2 protein appears to play a role in EICs in positive margins, and needs to be considered for avoiding positive margins in BCS on patients with breast carcinoma.

Adult↗

PubMedDate not supplied

Death receptor-dependent and -independent pathways in anticancer drug-induced apoptosis of breast cancer cells.

Apoptosis is a crucial event for anticancer drug efficacy. The signal pathways activated by anticancer drugs are classified as death receptor (DR) -dependent or -independent. The mechanisms by which anticancer drugs induce apoptosis via DR-dependent pathways are not fully understood. In the present study, we assessed differential activation of signal transduction pathways leading to apoptosis by anticancer drugs in breast cancer cell lines. Three breast cancer cell lines, MDA-MB-231, MCF-7, and MCF-7ADM, which is drug-resistant, were used. Drug sensitivity and apoptosis were assessed by MTT assay and TUNEL, respectively. Expression of apoptosis-related protein was assessed by Western blotting and RT-PCR. The sensitivities of MDA-MB-231 and MCF-7 cells to mitomycin C (MMC) and adriamycin (ADM) were similar. In contrast, sensitivity of MDA-MB-231 cells to paclitaxel (TXL) was 30-fold greater than that of MCF-7 cells, 0.03 micro M in MDA-MB-231 and 0.9 micro M in MCF-7 cells, respectively. Treatment with MMC increased expression of DR4 and Fas in a time-dependent manner in MDA-MB-231 cells. Treatment with ADM increased expression of DR4 and DR5 but not Fas, whereas treatment with TXL increased expression of Fas but not DR4 and DR5 in MDA-MB-231 cells. Although treatment of MCF-7 cells with ADM increased expression of DR4 and DR5 but not Fas, expression of DR4, DR5, and Fas by the drug-resistant cells did not change following treatment with ADM. Activation of Fas, DR4, and DR5 in drug-sensitive cells in response to anticancer drugs is dependent on the cytotoxic effect of each drug. In drug-resistant cells, apoptosis is induced via DR-independent pathways mediated by mitochondrial dysfunction.

Antibiotics, Antineoplastic↗

PubMedDate not supplied

Host-oriented peptide evaluation using whole blood assay for generating antigen-specific cytotoxic T lymphocytes.

A whole blood assay using antigenic peptide was established to predict host cytotoxic T lymphocyte (CTL) precursor status. Blood samples from HLA-A24 donors and colorectal cancer patients were directly diluted with RPMI-1640 medium to a 20% blood concentration, then distributed to tubes and a peptide of an HLA-A24-restricted CEA peptide panel (20 microM) was added to the tubes. Incubation was performed for 4-5 days and supernatants were subjected to ELISA specific for IFN-gamma protein. It was observed that certain CEA peptides could stimulate the diluted blood samples to produce IFN-gamma. Only the peripheral blood mononuclear cells (PBMCs) that were purified from the IFN-gamma-positive samples of the whole blood assay showed positive spots, detected with IFN-gamma ELISPOT assay, and could proliferate with the stimulation of immobilized anti-CD3 antibody plus interleukin-2 (CD3/IL-2 system). The proliferating PBMCs expressed cytotoxic activity against HLA-A24+ CEA-expressing tumor cells and the TISI target cells pulsed with the CEA peptide that had been used to stimulate the PBMCs to produce IFN-gamma, but they did not kill the target cells pulsed with peptides that had failed to stimulate IFN-gamma production, nor did they kill the target cells alone. Theses findings suggest that the IFN-gamma production of the blood samples detected by the whole blood assay identifies the peptide that can induce the CEA antigen-specific CTL response. Detection of IFN-gamma gene expression using real-time-PCR analysis could identify the peptide within 6 hours, which is earlier than the protein analysis by ELISA. The whole blood assay using the CEA peptide panel for healthy donors and colorectal cancer patients revealed that IFN-gamma-inducible peptides were different among the individual samples tested, indicating that the CEA peptides that should be used for generating CTLs are different in individual patients. The whole blood assay using a CEA antigen peptide panel is simple and beneficial for identifying candidate peptides. The host-oriented peptide evaluation (HOPE) approach may provide hope for the augmentation of clinical efficacies for peptide-based cancer immunotherapy.

B-Lymphocytes↗

PubMedDate not supplied

Enhancing effect of PS-K on IL-2-induced lymphocyte activation: possible involvement of antagonistic action against TGF-beta.

Effects of protein-bound polysaccharide (PS)-K on interleukin (IL)-2-induced responses of peripheral blood mononuclear cells (PBMCs) were studied. PS-K (50 mcg/ml) was observed to enhance proliferative responses, cytotoxic activities against K562 and Daudi target cells, CD25+ cell population and telomerase activity of PBMCs stimulated with IL-2. The cytotoxic effector cells could be generated in the presence of PS-K even with a minimum amount of IL-2. The enhancing effect of PS-K on the IL-2-induced lymphocyte activation was more evident in PBMCs from cancer patients than in those from healthy volunteers, suggesting that PS-K may be beneficial if combined in the IL-2-based immunotherapy of cancer. TGF-beta inhibited the IL-2-induced lymphocyte activation of proliferative responses, cytotoxic activities and CD25+ cell population, the inhibitions of which were abrogated with PS-K. PS-K also abrogated the TGF-beta-induced anchorage-independent growth of normal rat kidney cells. Flow cytometric analysis using a labeled TGF-beta revealed that PS-K blocked the binding of TGF-beta at its receptor level on the surface of PBMCs. It is suggested that PS-K enhances IL-2-induced lymphocyte activation through, in part, an antagonistic action against TGF-beta.

Adjuvants, Immunologic↗

PubMedDate not supplied

The ligands of peroxisome proliferator-activated receptor (PPAR) gamma inhibit growth of human esophageal carcinoma cells through induction of apoptosis and cell cycle arrest.

In the present study, we examined the expression of peroxisome proliferator-activated receptor gamma (PPARgamma) and the growth-inhibitory effects of Troglitazone and Pioglitazone, selective ligands for PPARgamma, using a series of human esophageal carcinoma cell lines (TE-1, -3, -7, -8, -12 and -13). PPARgamma expression was detected in all six human esophageal carcinoma cell lines. The esophageal carcinoma cell line TE-13 showed marked growth inhibition in response to Troglitazone and Pioglitazone. Flow cytometry performed on TE-13 cells exposed to Troglitazone showed that the cell cycle was arrested at the G1-phase. This result was confirmed by the finding of reduced cyclin D and cyclin E expression by Western blot analysis. DNA ladder formation was also detected, as was the induction of apoptosis-related proteins. Our results suggested that Troglitazone inhibited the growth of human esophageal carcinoma cell lines via G1 arrest and apoptosis and that PPARgamma ligands should be considered as possible target molecules in the treatment of human esophageal carcinomas.

Antineoplastic Agents↗

PubMedDate not supplied

Phase I study of combination therapy with S-1 and docetaxel (TXT) for advanced or recurrent gastric cancer.

BACKGROUND: S-1, an oral fluorouracil antitumor drug, and docetaxel have both been identified as effective agents for the treatment of gastric cancer. The two drugs have incompletely overlapping principal toxicities, which constitute the rationale for evaluating the effects of a combination of S-1 and docetaxel in this phase I study. The aim of this phase I study was to determine the maximum-tolerated dose (MTD) and the recommended dose of docetaxel with a fixed dose of S-1 in patients with advanced or recurrent gastric cancer. PATIENTS AND METHODS: The pharmacokinetics of both drugs were evaluated on Day 1 of treatment. Patients with a performance status (PS) of 0 to 2 received docetaxel at the starting dose of 40 mg/m2 by i.v. infusion over 1 hour on Day 1 and S-1 at the fill dose of 80 mg/m2 daily for two weeks every three weeks. Nine patients were treated with increasing dose levels of docetaxel as follows: (docetaxel/S-1, mg/m2): 40/80 (Level 1), 50/80 (Level 2) and 60/80 (Level 3) and all the cases were found to be assessable for drug safety, while 7 were assessable for response. Colony-stimulating factor (CSF) was not used in this study. The adverse effects of the treatment were analyzed according to NCI-CTC version 2, and the response was assessed according to the Japanese Classification of Gastric Cancer, 13th Ed. RESULTS: The MTD was reached at the 50/80 mg/m2 dose level in three patients out of six, who experienced a dose-limiting toxicity (DLT). The DLTs were neutropenia and allergic reactions. No hematological or non-hematological adverse effects (nore severe than Grade 2) were observed in any of the Level 1 patients. However among the Level 2 patients, 50% developed neutropenia (more severe than Grade 2), 33% developed loss of appetite, 17% developed diarrhea, 33% developed stomatitis and 17% developed allergic reactions. On the other hand, partial response was achieved in 5 (71.4%) of the 7 patients with evaluable lesions. The pharmacokinetics of docetaxel were not altered as compared to that in the historical controls by the administration of S-1. These results indicate that the recommended doses of the two drugs in the combination therapy would be 40 mg/m2 for docetaxel and 80 mg/m2 for S-1. CONCLUSION: The drug combination showed a good safety profile, with neutropenia being a common but manageable adverse reaction. Moreover, the responses observed in the study suggest that the drug combination shows a high degree of efficacy in patients with advanced and or recurrent gastric cancer.

Aged↗

PubMedDate not supplied

The spleen plays an immunosuppressive role in patients with gastric cancer: involvement of CD62L+ cells and TGF-beta.

CD62L is the human homologue of the murine lymphocyte homing receptor, mel-14. We investigated CD62L + cells in the spleen from patients with gastric cancer. Flow cytometric analysis revealed that CD62L + cells were decreased in the peripheral blood, but inversely increased in the spleen in parallel with disease progression in gastric cancer patients. The increased CD62L+ cells resided in the CD4+ suppressor-inducer phenotype, and the removal of CD62L+ cells from spleen cells resulted in a decrease of concanavalin-A-induced suppressor activity in vitro in one-way allogeneic mixed lymphocyte reaction. The CD62L+ cells included CD4+CD25+ regulatory T cells. The culture supernatant of CD62L + cells showed TGF-beta activity that permitted anchorage-independent growth of normal rat kidney (NRK) cells in a soft agar. TGF-beta activity was more significantly detectable in the splenic vein than in the peripheral blood, and TGF-beta mRNA was detectable in the spleen from advanced gastric cancer patients. These results suggest that CD62L+ cells migrate into the spleen with disease progression of gastric cancer and serve as suppressor-inducer cells with TGF-beta production to induce regulatory T cells, contributing to disease-associated immunosuppression in advanced gastric cancer patients.

Aged↗

PubMedDate not supplied

Experimental study on fluorescent microspheres as a tracer for sentinel node detection.

BACKGROUND: Common tracers for sentinel node navigation surgery are blue dye and technetium-99m-labelled colloids. However, in most esophageal or lung cancer patients, it is impossible to detect the sentinel node among mediastinal nodes by blue dye because of the anthracotic pigmentation of mediastinal nodes. The use of technetium-99m-labelled colloids requires a special facility, while a large hot-spot at the injection site prevents detection of the sentinel node around the primary lesion. To overcome these problems, we investigated the use of fluorescent microspheres (0.1-20 microm in diameter) as tracers in animals and detected fluorescence-positive nodes by a simple ultraviolet light irradiation method. MATERIALS AND METHODS: Two milliliters of fluorescent microspheres 0.1-20 microm in diameter diluted to 2.5% weight per volume was injected via the tail vein of 30 rats; systemic side-effects were examined. One milliliter of 1.0 microm-diameter microspheres dilution was injected on the backs of 30 rats; local side-effects were examined. A microsphere dilution (0.2 ml, 1.0 microm-diameter microspheres) was injected into the footpad of 18 rats; the lymphatic pathway and drainage were examined. Five milliliters of 1.0 microm-diameter fluorescent microspheres was injected endoscopically into the submucosa of the esophagus, stomach and small and large bowels of 6 domestic pigs, and 5 ml was injected into the subadventitia of the esophagus or subserosa of the stomach, and small and large bowels. Fluorescence-positive lymph ducts or nodes were carefully observed under ultraviolet light irradiation. RESULTS: No systemic side-effects were observed in rats. Only mild edema and a mild inflammatory reaction were observed on the backs of rats. Fluorescent microspheres 0.1, 0.5 and 1.0 microm in diameter were detected in lymph ducts or nodes of pigs within 1 hour after injection. CONCLUSION: Sentinel node navigation surgery with the use of fluorescent microspheres might be feasible and advantageous for patients with esophageal or lung cancer, especially in the mediastinum.

Animals↗

PubMedDate not supplied

HER2-specific cytotoxic activity of lymphokine-activated killer cells in the presence of trastuzumab.

We investigated whether trastuzumab, a humanized anti-HER2 monoclonal antibody, could induce HER2-specific cytotoxic activity on lymphokine-activated killer (LAK) cells. Trastuzumab alone was not toxic to the HER2-positive breast cancer cell lines MDA-MB453 and ZR75-1, nor to the HER2-negative breast cancer cell lines MDA-MB468 and MCF-7. LAK cells, which were activated with 1000 U/ml IL-2 for 4 days (4-day LAK), showed cytotoxic activity against the MDA-MB453, ZR75-1 and MCF-7 cells, but not against MDA-MB468 cells. LAK cell cytotoxic activity against the HER2-positive breast cancer cell lines MDA-MB453 and ZR75-1 was significantly augmented in the presence of 10 nM trastuzumab, but that against the HER2-negative breast cancer cell lines MDA-MB468 and MCF-7 was not. The cytotoxic activity of LAK cells plus trastuzumab against the MDA-MB453 cells was significantly inhibited by the addition of cold MDA-MB453 cells or cold ZR75-1 cells, but not by addition of cold MDA-MB468 cells. Twenty-nine percent of the 4-day LAK cells were CD16+, and the cytotoxicity of LAK cells plus trastuzumab was abrogated with the anti-CD16 antibody treatment of the LAK cells in the cytotoxicity assay. Only 7% of the 10-day LAK cells were CD16+, and the 10-day LAK cells failed to exhibit cytotoxicity even with trastuzumab. These results suggest that HER2-specific cytotoxic activity, which is mediated by an antibody-dependent cellular cytotoxicity (ADCC) mechanism, can be induced on LAK cells by the addition of trastuzumab.

Antibodies, Monoclonal↗

PubMedDate not supplied

Feasibility study of adoptive immunotherapy for metastatic lung tumors using peptide-pulsed dendritic cell-activated killer (PDAK) cells.

We have established a novel culture system to generate effector lymphocytes designated as peptide-pulsed dendritic cell-activated killer (PDAK) cells using cultured dendritic cells (DCs), synthetic peptide, peripheral blood lymphocytes, and interleukin-2 plus immobilized anti-CD3 antibody. A feasibility study of an adoptive immunotherapy trial using PDAK cells was conducted on HLA-A2 and HLA-A24 cancer patients with antigen-positive lung metastasis that was defined by serological analysis or PCR analysis. Eleven patients with lung metastasis participated in the study: 6 with colorectal cancer, 2 with pancreatic cancer, 1 each with breast and lung cancer, and 1 with melanoma. The patients received either Muc-1, CEA, gpl00, Her-2 or SART-3-PDAK cells generated in vitro, intravenously in combination with 350,000 U IL-2 weekly for 9 weeks, together with a planned dose-escalation schedule of three transfers each of 1 x 10(7), 3 x 10(7) and 1 x 10(8) PDAK cells/kg for 6 patients, and with a uniform dose of 3 x 10(7) PDAK cells/kg for the remaining 5 patients. Peptide/HLA-specific cytotoxic activity and TCRVbeta gene usage of PDAK cells were analyzed. All transfers of PDAK cells, which showed peptide/HLA-specific lysis, were well-tolerated in all patients, and adverse effects (elevation of transaminase, fever, and headache) were observed primarily at grade 1, but in no case greater than grade 2. The generation of sufficient cells to treat the patients with 3 x 10(7) PDAK cells/kg was feasible using our culture system, but we were able to generate and administer the dose of 1 x 10(8) PDAK cells/kg in only one patient. One partial response (PR) of lung metastasis occurred in a pancreatic cancer patient who received 3 x 10(7) Muc-1-PDAK cells/kg. The cytolytic units of PDAK cells in this patient appeared to be substantially higher compared to those in PD patients. TCR gene usage analysis on PDAK cells revealed preferential usage of TCRVbeta segments. These results suggest that adoptive immunotherapy using PDAK cells for cancer patients with antigen-positive lung metastasis is safe and feasible, and tumor response should be examined in a future clinical trial

Adult↗