We report the case of a 51-year-old female with stage IV advanced breast cancer accompanied by multiple bone metastases. A hard mass of about 3.0 cm in diameter was palpated just below the nipple. An excisional biopsy was performed and histological examination revealed infiltrated solid tubular adenocarcinoma. There were no estrogen or progesterone receptors in the tumor. Modified radical mastectomy was performed in October, 1998. Postoperative adjuvant therapy with 10 cycles of CEF therapy was undertaken for one year. Combined chemoendocrine therapy with 5'-DFUR and MPA was also conducted for 11 months. Bone scintigraphy showed that all bone metastatic lesions disappeared completely one year after the operation. Mild bone marrow suppression, alopecia and body weight gain were observed as side effects. It is suggested that this combination therapy may be useful for advanced breast cancer patients with multiple bone metastases.

The antitumor effect of cepharanthin (CR), a biscoclaurine alkaloid, was examined as to its direct action on tumor cells and inhibitory action on angiogenesis in tumors. The effect of CR on in vitro invasion by murine RL-[symbol: see text] 1 leukemia cells and Colon 26 tumor cells was studied using a biocoat matrigel invasion chamber. One hundred micrograms/ml of CR inhibited tumor cell invasion. Early induction of apoptosis was assayed by the binding of annexin V and phosphatidylserine (PS) in the cellular membrane CR (10 and 100 micrograms/ml) induced apoptosis in human Daudi and Raji B lymphoblastoid cells. Treatment with CR (1 and 10 micrograms/ml) also inhibited the in vitro growth of Daudi and Raji cells. Ten micrograms/ml of CR also inhibited the growth of human umbilical vein endothelial cells (HUVEC) and human dermal microvascular endothelial cells (HMVEC). These results indicate that CR has diversified antitumor functions, i.e., an enhancement of a sequential immune mechanism, a direct cytotoxic effect and inhibitory action of angiogenesis in tumors.

The purpose of this study was to investigate the correlation between efficacy and dose intensity of postoperative adjuvant chemotherapy with MMC and UFT. A total of 1,410 patients from 180 institutions were allocated into a low-dose group and a high-dose group. The patients in the low-dose group received MMC at 8 mg/m2 on the day of surgery and 3 capsules of UFT (300 mg in tegafur) daily for 6 months. The patients in the high-dose group received MMC at 8 mg/m2 on the day of surgery, and in weeks 4, 10, 16, and 22 after surgery and 6 capsules of UFT (600 mg in tegafur) daily for 6 months. The patients in the high-dose group tended to exhibit higher survival rates than those in the low-dose group, although the difference was not significant. For the n(+)ps(-) patients, however, the survival rates were significantly higher in the high-dose group (p = 0.043). The recurrence-free rates showed a similar tendency. The incidence rates of adverse events were significantly higher in the high-dose group than in the low-dose group. Compliance was poorer in the high-dose group. Although the number of adverse events increases, a better prognosis can be expected with a high dose. These results confirmed a dose-dependency in adjuvant chemotherapy with MMC and UFT.

The occurrence of delayed emesis induced 24 h after the administration of a non-platina chemotherapeutic agent, doxorubicin hydrochloride (doxorubicin), as well as behaviors such as feeding, drinking and defecation were examined in dogs. A single intravenous administration of 2 mg/kg doxorubicin induced emesis within 24 h of administration in some dogs, while delayed emesis was observed 24 h after administration in all dogs. This delayed emesis emerged strongly at day 3 or 4 and decreased at day 5. Hypophagia, the decreased frequency of drinking and the increased frequency of defecation were induced shortly after delayed emesis. Twenty-four hours after the administration of doxorubicin, a daily dose of 0.3 and 1 mg/kg/day, p.o. azasetron, a 5-HT3 antagonist, was administered for 4 days. Doxorubicin-induced delayed emesis was observed to decrease by about 30 and 50%, respectively. This result suggests that 5-HT3 receptors play a role in the mechanism of delayed emesis. Azasetron was found to improve the increased frequency of defecation, but exerted no obvious effect on hypophagia or on the decreased frequency of drinking. Taken together, we suggest that doxorubicin-induced emesis in dogs is a useful method to study further the mechanisms of delayed emesis and to investigate novel therapeutic agents against delayed emesis.

The anti-cancer effect of 5-fluorouracil (5-FU) is significantly affected by the intratumoral environment. Elevated expression of thymidylate synthase (TS), the target enzyme of 5-FU, and a lack of reduced folate or FdUMP results in insufficient inhibition of TS. Further, elevated expression of DPD in the tumor tissue results in a lack of FdUMP. TS-1, which contains tegafur and dihydroxypyridine (CDHP), a potent DPD inhibitor, is a promising anticancer drug. Isovorin has been available in Japan since autumn 1999. In cancer patients, folate deficiency may be derived from increased consumption and/or absorption disturbance of folate. In such situations, sufficient TS inhibition cannot be obtained if 5-FU is administered without supplementation of reduced folate. We describe in detail the metabolism of those substances in relation to the anticancer effect of 5-FU. Further, we show the theoretical construction of first-line and second-line chemotherapy with consideration of TS and DPD expression.

We investigated whether 5-fluorouracil (5-FU) can induce apoptosis in breast cancer cells. Preoperatively, 23 breast cancer patients were divided into a group treated with 5-FU at 200 mg/day for 2 weeks (Group A) and a non-treatment group (Group B), and breast cancer tissues were taken postoperatively. DNA fragmentation by agarose electrophoresis and the TUNEL method were used to investigate the induction of apoptosis. The labeling rate with Ki-67 was measured to study the reproductive activity of tumor cells. The involvement of p53 in the apoptosis decision mechanism was also studied. DNA was more fragmented in Group A than in Group B. The apoptosis index by the TUNEL method was 1.88 +/- 1.03 in Group A, which was significantly higher than 0.36 +/- 0.86 in Group B. The labeling rate with Ki-67 was significantly higher in Group B(62.3 +/- 21.7) than in Group A (29.8 +/- 16.0). There was no difference in the protein expression of p53 regardless of the presence or absence of DNA fragmentation. These results indicate that 5-FU administration induces apoptosis in breast cancer cells and significantly inhibits their reproductive activity. Involvement of p53 in the apoptosis decision mechanism was not demonstrated.

In our attempt to find characteristic genetic changes in resistant tumors, we screened the whole genome for gene aberrations in 28 primary ovarian cancers, using the comparative genomic hybridization (CGH) method. These cancers included 14 tumors from patients who did not respond to cisplatin-based combination chemotherapy in comparison with 14 tumors from patients who completely responded to the chemotherapy. We found gains in chromosomal region 1q21-q22 and 13q12-q14 to be related to the drug-resistant phenotype in ovarian cancer patients. Several genes encoding transcription factors, oncogenes, cell cycle regulators and regulators of the apoptotic pathway are found to be located on these regions of the chromosomes, and these genes are potential modulators for toxic insults in cancer cells. This is the first report that shows the relationship between certain genomic aberrations and clinical resistance for cisplatin-based chemotherapy in ovarian cancer patients based on the CGH analysis. Present findings suggest that these chromosomal gains may be potential indicators for prediction of resistance in ovarian cancer patients prior to cisplatin-based chemotherapy.

It has not clearly been elucidated how differently differentiation-inducing drugs act on tumor cells, whether they promote differentiation or apoptosis. To elucidate the mechanisms whether leukemic cells responding to ONO-4007, a lipid A derivative, undergo differentiation or apoptosis, we established two cell clones from a rat myelomonocytic leukemia c-WRT-7/P2 clone which undergoes differentiation followed by apoptosis by ONO-4007-treatment. One of the clones (1D6) showed the features of differentiation, such as phagocytosis when treated with ONO-4007 more than 24 hrs. The other clone (3B1) clearly showed the features of apoptosis, such as DNA ladder formation within 24 hrs after incubation with ONO-4007. We then examined expression of CD14, p21, p38MAPK, JNK/SAPK, and bcl-2, functional p53 statuses and cell cycle in these two clones, and revealed the following: Without treatment with ONO-4007; 1) CD14, p21, and bcl-2 proteins were equally expressed in both clones; 2) wild-type and non-functional mutated-type p53 were present in both clones and the p53 in 3B1 clone was recessive whereas that in 1D6 clone was dominant negative; 3) p38MAPK in 3B1 clone was already phospholyrated whereas that in 1D6 clone was not. After treatment with ONO-4007; 1) neither expressions of CD14 nor that of p21 protein was changed in any of the clones; 2) p38MAPK in 3B1 clone was dephospholyrated at 1 and 2 hrs after treatment whereas that in 1D6 clone was phospholyrated at 4 and 8 hrs after treatment; 3) the expression of bcl-2 protein in 3B1 clone was reduced. These findings suggest that p53 may be one of the key factors in leading these cells to differentiation or apoptosis, and that bcl-2 may suppress the apoptosis.

An 11-year-old boy was diagnosed as having acute lymphoblastic leukemia (ALL, L1) in 1987 and underwent treatment with an ALL high-risk protocol (prednisolone, vincristine (VCR), daunorubicin, 1-asparaginase), which resulted in complete remission. In 1990 he developed chronic hepatitis C and received interferon therapy. In December 1994, ALL recurred, and the patient was treated with VCR. He subsequently developed severe hemolysis (Hb 12.5 g/dl-->6.8 g/dl) with increases of indirect bilirubin, AST, and LDH. Furthermore, symptoms resembling a syndrome of inappropriate secretion of ADH (SIADH) and DIC developed. Upon incubation of the patient's red blood cells with VCR in vitro, extreme deformity of the cells was observed. These findings suggested that splenomegaly, due to liver cirrhosis which had developed rapidly from chronic hepatitis C while the patient was in an immunosuppressed state induced by anticancer drugs, had trapped the deformed red blood cells and resulted in severe hemolysis. The patient died on the 165th day after admission due to liver failure.

A 62-year-old man was diagnosed as being in the chronic phase of chronic myelogenous leukemia (CML) in 1990, and subsequently treated with hydroxyurea (HU). The total HU dose administered reached 2,929 g (average, 1.44 g/day). In December 1995, the patient was admitted to our hospital for control of the CML. Following HU therapy, he often experienced high fever (38-39 degrees C) due to infection or blastosis, and at that time his skin showed marked pigmentation, dryness and scaling with itching and anhidrosis. A skin biopsy sample from the left scapula showed atrophic change of the skin and epidermal tissues with fibrotic changes and damage to the subcutaneous glands. This was strongly suspected to have been caused by the continuous HU administration, and the anhidrosis and dryness was considered to have contributed to the patient's high body temperature. Frequent cooling of the patient's body was effective.

Molecular mechanisms of DNA recognition and functional expression by bioactive compounds such as bleomycin, enediyne antibiotics, and zinc finger proteins are an important research subject in the pharmaceutical scientific field. Cleavage of cellular DNA by bleomycin substantially contributes to the antitumor activity of this drug. Some physicochemical data have clearly demonstrated that the bleomycin-iron (II) complex forms a dioxygen adduct species. Of special interest is the fact that the bleomycin-iron complex and cytochrome P450 have a similar dioxygen activation cycle. Probably, the site-specific iron-peroxide species of bleomycin accounts for the action mechanism of selective DNA cleavage by bleomycin. The enediyne antibiotics possess an unprecedented chemical structure, potent anticancer activity, and a fascinating mode of biological action. A new compound, C-1027, consists of a carrier apoprotein and an enediyne chromophore. The high-resolution NMR experiments clarified that novel chromophore interacts with DNA through its benzoxazolinate and aminosugar moiety, and also with apoprotein through the benzoxazolinate and macrocyclic moiety. The results provide a molecular basis for the host-recognition mode, the reaction mechanism, and the drug delivery system of chromoprotein C-1027. In addition, we found unique reactivity of C-1027 chromophore toward a tRNA(Phe). RNA as well as DNA might be also a potent biological target of the enediyne antibiotics. On the basis of characteristic DNA binding mode of Cys2His2-type zinc finger motif, artificial zinc finger proteins have been created, and new functions such as DNA cleavage, long sequence binding, and DNA bending are produced. The first artificial His4-type zinc finger protein is also made from Cys-->His mutations of the Cys2His2-type transcription factor Sp1. Such novel zinc finger proteins may be useful as a gene therapeutic agent and a tool for genetic engineering.

We developed chitin-containing cisplatin (CDDP) albumin microspheres (MS). We also investigated physical properties of MS in vitro, anti-tumor effect of MS in VX2 tumor model rabbits in vivo and clinical evaluation of MS. The CDDP contents, specific surface areas and yield of MS increased with an increase in the concentration of chitin. These findings suggest that accumulation of chitin on the surface of microsphere was the cause of expansion of the coated area as the concentration of chitin increased. In vitro CDDP releases decreased as the concentration of chitin increased. The initial burst effect of the drug was to be controlled by the increase of concentration of chitin. The chitin-containing CDDP microspheres showed similar CDDP release patterns irrespective of the concentration of chitin between 1 and 6 h after the administration of CDDP microspheres. After 6 h, the blood Pt levels increased with an increase in the concentration of CDDP microspheres suggesting a significant effect of the concentration of chitin on microsphere decomposability in vivo. Tumor growth rate was increasingly suppressed as the concentration of chitin increased. Clinical findings indicated that antitumor activity of chitin-containing CDDP albumin microspheres would have a chemoembolic effect by embolizing the hepatic artery. In the tumor and necrosis portions phagocytosis of microspheres by macrophage-like giant cell was shown. Consequently, the biocompatibility and decomposable properties of chitin-containing CDDP albumin microspheres were demonstrated.

Paclitaxel has been reported to be effective for the treatment of CDDP resistant tumors. Thus, the efficacy of paclitaxel on CDDP resistant HEp-2 and KB head and neck squamous carcinoma cell lines was evaluated in monolayer and multicellular tumor spheroids (MTS). Cell lines with a tenfold resistance to CDDP were used in this study (Tanaka, K. et al, Keio J Med, 70. 1993). MTS were developed using the liquid overlay culture technique. After exposure to graded concentrations of drugs and different exposure time, the cells were subjected to a clonogenic assay. The effect of paclitaxel on both monolayer and MTS was dependent on the drug concentration and related to the exposure time. For HEp-2 MTS, 10(-7) M/L of paclitaxel resulted in a cell death rate of approximately 90% in both parent and resistant cells. For KB MTS, the cells were more resistant to paclitaxel than the HEp-2 cells, and a 72 hour exposure time was needed to achieve a cell death rate of approximately 90%. These data suggest that paclitaxel may be effective for treating CDDP resistant head and neck cancer.

A 58-year-old man had been treated with one intravenous injection of 120 mg of nimustine hydrochloride (ACNU), ten thrice-weekly doses of 3,000,000 U of interferon beta, and brain irradiation for cerebral glioblastoma. One month later he had fever, appetite loss, a productive cough and dyspnea. Chest radiography and CT showed diffuse, nonsegmental ground glass opacity in both lung fields. Hypoxemia and lung shadows were exacerbated day by day. Bronchoalveolar lavage revealed an increases in the total cell count and the percentages of lymphocytes and neutrophils, and a decrease of the CD 4/8 ratio. Interferon beta therapy was stopped, and steroid pulse therapy and prednisolone 40 mg administration were initiated. The symptoms, hypoxemia and lung shadows quickly improved. Reported cases of interferon beta-induced pneumonia are rare.

The adverse effects of combination chemotherapy with paclitaxel and carboplatin, including neurotoxicity, arthralgia and muscle pain, were evaluated in 21 patients (30 courses) using questionnaires of the Gynecologic Oncology Chemotherapy Joint Research Group. The scores of pain and numbness peaked from the third to fourth day of treatment. Although the pain score improved subsequently, the numbness score persisted at a high level. Compared to the first and second courses, the peak pain score was higher and persisted for a longer duration in the fifth and sixth courses. Using the questionnaires, we were able to recognize a high incidence of numbness and pain in patients on combination chemotherapy with paclitaxel and carboplatin, and identify the degree and temporal changes of the adverse effects. Our results suggest that the questionnaires used in this study are clinically useful for evaluating the degree and clinical course of pain and numbness in anticancer chemotherapy.

The pharmacokinetics of CPT-11 administered into rats to evaluate the future possibility of i.p. administration was investigated. Serum, bile juice, and intraperitoneal fluid was collected to measure the concentration of CPT-11, its metabolite SN-38, and SN-38 glucuronized (SN-38 glu). The concentration of CPT-11 was elevated after CPT-11 administration into the peritoneal cavity. The serum and biliary concentrations of SN-38 and SN-38 glu were increased and prolonged shortly after i.p. administration of CPT-11. These results suggest the possibility of i.p. administration of CPT-11 in the future.

Vinorelbine (VNR) is a new vinca alkaloid derivative semi-synthesized by Potier et al. The antitumor activity of VNR was superior to other vinca alkaloid antitumor agents, and the neuro-toxicity of VNR was weaker than those of other vinca alkaloids. In nude mice xenografted human tumor models, VNR showed antitumor activity against eight of eleven tumor models (non-small cell lung cancer: 4/4, breast cancer: 2/3, colon cancer: 0/2, stomach cancer: 2/2). Especially, VNR showed tumor-regressive activity against LC-6 non-small cell lung cancer and MX-1 breast cancer. The antitumor activity of VNR against non-small cell lung cancer was superior to that of vindesine (VDS), which had been one of the key drugs of non-small cell lung cancer in the clinic. In combination chemotherapy, VNR plus cisplatin (CDDP) was better than VDS plus CDDP, which had been one of the standard regimens of non-small cell lung cancer chemotherapy. The potent antitumor effect of VNR with minor neurotoxicity was explained by VNR having stronger activity on mitotic microtubules than axonal microtubules. It was supposed that less activity of VNR against mitotic microtubules would be related to different composition of microtubule-associated TAU isoforms in the two types of microtubules. In non-small cell lung cancer, VNR resulted in a significantly higher response rate than VDS. In combination with CDDP, VNR resulted in longer survival than VDS with a significant log-rank test. In advanced breast cancer, VNR resulted in a high response rate in 1st line and 2nd line treatment. VNR is effective in combination with chemotherapeutic agents such as anthracycline, fluorouracil and Taxol. In Japan, the clinical trial in breast cancer is now ongoing.

LN382, a glioblastoma cell line, has a temperature-sensitive mutant p53. At the permissive temperature (34 degrees C), arrest of cell growth at the G1 phase occurred with recovered p53's transcriptional activity, and restored p53 protein turnover. In order to understand the influence of the functional status of p53 on the sensitivity to anticancer agents in glioblastoma cells, I analyzed responses of LN382 cells and U251MG cells with a mutant p53 as a control at 34 degrees C and 37 degrees C to etoposide, paclitaxel, and cisplatin, and 1-(4-amino-2-methyl-5-pyrimidinyl)methyl-3-(2-chloroethyl)-3-nitrosourea hydrochloride (ACNU). Restoration of p53 protein function in LN382 cells at 34 degrees C reduced the cytotoxicity of etoposide and paclitaxel, whereas that of cisplatin and ACNU was not affected. In contrast, the temperature shift to 34 degrees C did not alter the cytotoxicity of etoposide, paclitaxel, cisplatin, or ACNU in U251MG cells. Transduction of wild-type p53 in LN382 cells also reduced the sensitivity of the cells to etoposide. Cell cycle analysis using flow cytometry revealed that this decrease in sensitivity was associated with an impaired transition to the G2M phase subsequent to the addition of etoposide or paclitaxel. These results indicate that cell cycle arrest induced by wild-type p53 function may abrogate the cytotoxic effects of etoposide and paclitaxel, which are dependent on G2M-associated apoptosis.

Depression is well known as one of the psychiatric complications of interferon (IFN) therapy in patients with chronic active hepatitis C. We compared the psychiatric status during interferon therapy between patients with renal cell cancer (RCC) and those with chronic active hepatitis C.