The effect of anticancer drugs on invasive capacity of Lu 135 human small-cell lung cancer cells was studied in vitro. the invasive capacity decreased in a dose-dependent manner when tumor cells were treated with the anticancer drugs cisplatin and etopside. The inhibition of tumor cell invasion was almost parallel with the inhibition of tumor cell growth. The anticancer drugs also suppressed tumor cell migration and the activity of the matrix-degrading protease, type IV collagenase, activity. But they did not suppress adhesion to basement membrane protein such as laminin, fibronectin, or type IV collagen. Because tumor cells express adhesion molecules on the cell surface, the effect of the anticancer drugs on the expression of one such molecule, integrin, was also studied. Lu 135 cells expressed alpha 4/beta 1 integrin on the cell surface, and the pattern of integrin expression did not change with exposure with the anticancer drugs. These data suggest that the anticancer drugs inhibit the invasive capacity by suppression of migration and type IV collagenase activity of tumor cells, and that they do not modulate adhesion to the extracellular matrix or cell surface adhesion molecules such as integrin. Furthermore, these findings indicate that anticancer drugs may be useful for anti-metastatic therapy in patients with small-cell lung cancer.

To evaluate the difference of anticancer drug sensitivities of cancer cells among altered cancer cell-populations in the MTT assay, we compared the results of MTT assay of cancer cells obtained by the conventional technique and of the cancer cell rich fraction from centrifugal elutriation (CE) system in 17 tumors and 3 peritoneal and pleural aspirates from 20 gastric cancer patients. CE raised the population of the cancer cells from 36.1% to 75.4% in tumors and from 37.6% to 94.9% in aspirates respectively. And the rate of yield of cancer cells after CE was 51.9% in tumors and 61.9% in aspirates. In 10 cases, anticancer drug sensitivities of cancer cell rich fraction were different from those of cancer cell obtained by the conventional technique, and mostly in these cases the population of the cancer cells before purification by CE was less than 30%. Pathologically, mostly in undifferentiated type, severe infiltrated type, and scirrhous type carcinoma the population of the cancer cells before purification was less than 30%. Therefore, in these cases, cancer cell rich samples should be used in determining the correct chemosensitivity of cancer cells.

A 64-year-old woman with multiple myeloma, IgG lambda type Durie-Salmon Stage II, was admitted because of gradually developing anemia and increased blasts with abnormal karyotype in her bone marrow after 10 years of treatment. The chromosomal analysis showed 44, XX, del(5q), del(7q), -9, add(12p), -21, typical of secondary MDS due to the cumulative alkylating agents. Thrombocytosis concomitantly occurred with emergence of chromosomal abnormality, but the serum interleukin 6 level was not elevated, which suggested that it was related to development of secondary MDS.

Resistance of tumors to a variety of chemotherapeutic agents presents a major problem in cancer treatment. The gene responsible for multidrug resistance, termed mdr1, encodes a membrane glycoprotein (P-glycoprotein) that acts as a pump to transport various cytotoxic agents. The P-glycoprotein has been shown to bind anticancer drugs and several resistance-reversing agents including calcium channel blockers, and to be an ATPase. The P-glycoprotein was found to function in the blood-brain barrier. The physiological function of the P-glycoprotein in relation to therapy is discussed.

A new human endometrial adenocarcinoma cell line, Sawano cells, was established from an endometrial adenocarcinoma from a 61-year-old woman and has been maintained in vitro for more than 2 years and 10 months. The cells were found to form a monolayer in a mosaic fashion and to tend to pile up. Population doubling time was 43.2 hours at the 10th generation. The modal chromosomal number of the cell fell in a diploid range. Histology of the tumor induced in athymic mice showed it to be a moderately differentiated adenocarcinoma which closely resembled the original human tumor. Estrogen and progesterone receptors were not demonstrated in the in vitro culture cells and in the tumors induced in nude mice, though they were positively demonstrated in the original tumor. The cells had intrinsic cisplatin-resistance (50% inhibition concentration:6.63 micrograms/ml) at 120 hours of contact. We believe this cell line with help us to investigate the biological mechanisms of CDDP resistance.

Sixteen patients with refractory small cell lung cancer (SCLC) were treated with an irinotecan starting dose of 100 mg/m2 given as a 90-minute iv infusion every week with subsequent doses based on toxicity. Mean age was 64 years; 14 male and 2 female; 4 with limited disease and 12 with extensive disease; all patients pretreated with combination chemotherapy containing etoposide. The overall response rate was 50% (95% CI 25-75%) with no CR and 8 PR. The median duration of response was 46 days. Major toxicities were leukopenia, diarrhea and pulmonary toxicity. Irinotecan was thus an effective agent in refractory SCLC.

Tilivalline (1a), a metabolite isolated from Klebsiella pneumoniae var. oxytoca, belongs to a group of pyrrolo[2,1-c][1,4]benzodiazepines, a characteristic skeleton of anthramycin-type antitumor antibiotics. We have accomplished a completely stereoselective, efficient and convenient synthesis of 1a utilizing a new Mannich type intramolecular cyclization as a key step. Further, a computational chemical analysis clarified the effect of zinc chloride on the high stereoselectivity in the tilivalline synthesis. To aim both the extension of the scope of the new Mannich type intramolecular cyclization and the studies on the structure-biological activity relationship, we further extended the method to the synthesis of tilivalline derivatives and 2-(3'-indolyl)-1,4-benzodiazepines (50). Investigation on the cytotoxicity of 1a and its analogs has revealed that 1a shows the strong cytotoxicity toward mouse leukemia L 1210 cells and the replacement of the indole function of 1a with cyano one increases the cytotoxicity of 1a about 100 times (IC50 = 0.05 microgram/ml).

We performed primary immature Sertoli cell culture to investigate whether or not anti-cancer agents would have a direct effect on rat Sertoli cells. Sertoli cells, isolated from testes of 18-day-old rats, were cultured in pellets with medium for 5 days. The concentration of transferrin in cultured medium were measured as the function of Sertoli cells. The anti-cancer agents cis-diamminedichloroplatinum (CDDP), adriamycin and vinblastine were selected for this study, and added to the culture medium. CDDP decreased the level of transferrin concentration in cultured medium, namely 0.5 microgram/ml of CDDP resulted in 54.9% of the transferrin concentration in the medium compared with that without any anti-cancer agents (p < 0.05), and 1.0 micrograms/ml of CDDP produced transferrin concentrations of 57.5% and 46.2% (p < 0.05), respectively. Adriamycin (0.4 microgram/ml) and vinblastine (0.5 microgram/ml) produced transferrin concentrations of 35.2% and 31.3% in cultured medium (p < 0.05), respectively. These findings revealed that anti-cancer agents have direct damaging effects on rat Sertoli cells.

We investigated the spermatogenic function of patients with testicular cancer, and the influences of anti-cancer chemotherapy on testicular function in these patients. Fifty-one patients with testicular cancer were selected for evaluation of their testicular function, including spermatogenesis and endocrinological function, before and after chemotherapy with anti-cancer agents. Before chemotherapy with anti-cancer agents, 22 of 49 patients (44.8%) had a sperm concentration of less than 20 x 10(6)/ml, and 8 patients (16.3%) showed azoospermia. The mean sperm concentration of the patients with testicular cancer was 29.0 x 10(6)/ml before therapeutic chemotherapy with anti-cancer agents, but within 3 months after chemotherapy, it decreased to 3.86 x 10(6)/ml (p < 0.01). Fifteen of 19 patients (73.7%) were revealed to have azoospermia. Damage to spermatogenesis became more severe with the number of chemotherapy treatment. No patients had a detectable sperm count at the completion of 3 or more courses of chemotherapy. But some patients who received 3 courses or more of chemotherapy showed recovered sperm counts after 2 or more years. Thus, a lack of sperm after chemotherapy for cancer did not necessarily indicate inability to recover spermatogenesis over 2 years after chemotherapy. Serum FSH levels of the patients were 5.62 +/- 3.43 mIU/ml before chemotherapy, and 19. 70 +/- 17.06 mIU/ml (p < 0.05) at the time of its completion. Serum FSH levels could reflect damage to spermatogenesis in these cases. Cases in which spermatogenesis did not recover may have higher serum FSH levels than those with recovery of spermatogenesis.

We investigated the mechanisms of CPT-11-induced diarrhea. 1) CPT-11 (80 mg/kg, i.v.) induced watery diarrhea within 1 hr after dosing in saline-loaded (10 ml/kg, p.o.) rats. This was partially inhibited by subcutaneous injection of atropine (1 mg/kg) or ondansetron (1 mg/kg) and almost completely inhibited by a combination of atropine and ondansetron or by clonidine (0.3 mg/kg) or morphine (10 mg/kg) alone. 2) CPT-11 at the same dose reduced intestinal fluid absorption, which was blocked by the anti-diarrheal agents mentioned above. Intraluminal injection of CPT-11 (20 mg/2 ml) inhibited fluid absorption and induced fluid secretion. 3) CPT-11, 60 mg/kg, by single intravenous injection induced fewer enzymological and histological changes in the small intestine than 5-FU at 270 mg/kg, while 4 consecutive dosings of CPT-11 induced delayed diarrhea (days 5-7) associated with disruption of intestinal integrity. Co-administration with anti-diarrheal agents, except for ondansetron, protected against watery diarrhea appearing within 1 hr after CPT-11 on days 3 and 4, but worsened delayed diarrhea. These results suggest that single injection of higher doses of CPT-11 causes watery diarrhea at an acute phase at least partly by reducing fluid absorption or increasing secretion, and that while conventional anti-diarrheal agents protect against watery diarrhea, their co-administration in repeated CPT-11 administration has no ameliorative effect on CPT-11-induced delayed diarrhea.

Combined effect of bleomycin (BLM) and N-solanyl-N, N'-bis (3, 4-di-methoxy-benzyl)-ethylenediamine (SDB-ethylenediamine), a synthetic isoprenoid was studied on 3 oral squamous carcinoma cell lines (SCCTF, SCCKN, SCCRY) and clonal cell lines from SCCRY in vitro and on transplantable nude mouse tumors. The cytotoxicity of SDB-ethylenediamine varied among those cell lines in vitro. SDB-ethylenediamine potentiated more than 2-fold the cytotoxic effect of BLM in 6 cell lines, but no distinct correlation was found between cytotoxicity of SDB-ethylenediamine and potentiation by BLM. Compared with verapamil, the cytotoxic effect of BLM was potentiated 3.9-fold by SDB-ethylenediamine in SCCTF. The growth of the transplantable nude mouse tumors (SCCRY and SCCTF) was strongly suppressed when BLM was combined with SDB-ethylenediamine. Correlation between BLM resistance and potentiation of BLM by SDB-ethylenediamine was not observed in this experiment.

Twenty-six patients with cerebral malignant glioma (7 cases of astrocytoma Grade 3, and 19 of astrocytoma Grade 4) were treated by intra-arterial and local administration of MCNU. Nineteen patients were treated in combination with local radiation in a dose of 60 Gy. Intra-arterial administration of MCNU was performed by puncture of the ipsilateral common carotid artery and injection of 25mg of MCNU in 20 ml of physiological saline. Local administration of MCNU was performed by puncture of the Ommaya reservoir placed within the cavity after tumor resection. Objective tumor regression was observed on computerized tomography (CT) scans after intra-arterial and/or local administration of MCNU combined with radiotherapy in three of seven patients who had evaluable enhanced lesions on CT after surgery. It was also observed after chemotherapy alone in one of three patients with evaluable lesions. The response rate was 42.9% among patients treated with MCNU in combination with radiotherapy, and 33.3% in patients treated with MCNU alone. In two patients, local administration of MCNU induced brain edema, which was transient and caused no neurological sequelae. One patient suffered mild thrombocytopenia after seven intra-arterial doses of MCNU, however, no myelosuppression requiring administration of gamma-GCSF or blood transfusions was observed. These findings suggest that intra-arterial and local administration of MCNU can be expected to serve as effective and non-myelosuppressive chemotherapy in patients with cerebral malignant gliomas.

In SDI test using tetrazolium salt, we compared MTT with XTT, which has been recently developed and is more sensitive than MTT. Further, a chemosensitivity test with endoscopic biopsy specimens using XTT was conducted. The reproducibility of the MTT assay was assessed under various conditions. The I.I of MTT assay did not vary according to type or concentration of anticancer drugs and the biopsy site of specimens. Then MTT and XTT assay were compared, revealing the results of these 2 assays were significantly correlated. Thus, XTT assay was performed with tissues from surgical specimens using biotome. Of the 16 specimens subjected to XTT assay, 6 showed an OD value of 0.100 or more, which was well within assessment. XTT assay was also performed in biopsy specimens from patients using a biotome manipulated under endoscopic guidance before operation. Three among 6 specimens showed an OD value of 0.100 or more. Our findings have demonstrated that the SDI test using XTT can be used for the chemosensitivity test, even when these specimens contain a relatively small number of cells collected using a biotome.

The concentrations of carmofur, 1-hexyl-carbamoyl-5-fluorouracil (HCFU), in the serum, bile, pancreatic juice and pancreatic tumor tissue were studied in 18 cases of peri-pancreatic head cancer with drainages of the pancreatic and biliary ducts after pancreatico-duodenectomy. As a result, high concentrations of HCFU and 5-FU were detected in the serum, bile and pancreatic juice after a per-oral administration of HCFU, 200 mg: HCFU was high in the order of serum > bile and pancreatic juice, and 5-FU in the order of bile, serum and pancreatic juice. Two hours after administration, 5-FU concentration in the bile and serum attained to the maximum levels of 0.45 and 0.19 micrograms/ml, respectively, which demonstrated a lasting transfer of 5-FU to the bile at high concentration. HCFU and 5-FU levels in the tumor tissue were 0.079 and 0.024 micrograms/g, respectively. In conclusion, antitumor effect against the malignant tumors of the pancreatobiliary system can be expected by peroral administration of carmofur.

Head and neck cancer has hypoxic compartment. The hypoxic cells are resistant to anticancer drugs and thought to be one of the causes of recurrence. We examined effects of anticancer drugs on the hypoxic cells using HEp-2 human laryngeal cell line. Anticancer drugs, CDDP, peplomycin (PEP), 5-FU, THP-adriamycin (THP-ADR), and adriamycin (ADR), were incubated with cells in hypoxic condition (5% CO2 and 95% N2) for 72 hours. Drug effects were measured by proliferation rates (IC50). IC50 of PEP increased 22.89 folds in hypoxic cells. IC50 of CDDP and 5-FU also increased 1.86 and 2.27 folds respectively. However, IC50 of THP-ADR and ADR remained same in the hypoxic cells. It was proved that THP-ADR and ADR were effective for the hypoxic cells. The combination effect with THP-ADR and CDDP or PEP were more than additive against the hypoxic cells. It was suggested that the combination with THP-ADR and CDDP or PEP are effective for head and neck cancer which contains hypoxic cell compartment.

Toxicity criteria used by the "Japan Society for Cancer Therapy" (JSCT) were reviewed and compared with that of WHO, ECOG, NCI (USA), and JCOG. During the past ten years, these toxicity criteria of the major cooperative study groups have undergone significant revision, but, toxicity criteria of JSCT have never been revised for more than ten years. Thus criteria of JSCT now show a several inconsistency. Grading of toxicity is also indistinct. Therefore, toxicity criteria used by JSCT should be revised. Furthermore, in the assessment of toxicity, duration of toxicity, effects of toxicity on the quality of life and psychosocial problems should also be considered. Standard criteria for the assessment of long-term complications are necessary. To study early signs and symptoms of late effects and to know risk factors for toxicity are also important.

In the pre-clinical study of anticancer candidate compounds, the following three points should be carefully evaluated before decision to enter the clinical trials. Namely, 1) the data on the antitumor efficacies (in vitro and in vivo), 2) animal toxicities and 3) pharmacological studies. Since more attention has been placed on the clinical trials to protect the right of cancer patients, the toxicity data are especially examined carefully using small and large experimental animals. Feasibility to enter in the clinical studies should be examined by above three points in the preclinical studies.