We studied an antiproliferative effect of cancer chemotherapeutic agents, interferon (IFN) and, in particular, their combination effect on renal cell carcinoma. Either of colony formation assay (CFA) or [3H]-thymidine incorporation assay ([3H]-TdR assay) was employed as an in vitro chemosensitivity testing system. When compared, these two systems produced similar results with a good correlation (r = 0.97, p less than 0.01), in the antiproliferative effect of 30 drugs for 4 primary renal cell carcinomas and 5 xenotransplantable renal cell carcinomas. In vitro chemosensitivity test (CFA or [3H]-TdR assay) screened successfully only 5 "sensitive" drugs (7.9%) out of a total 63 cancer chemotherapeutic agents for 24 human renal cell carcinoma. This confirmed the findings reported by others. In the study of the antiproliferative effect of a cancer chemotherapeutic agent, human lymphoblastoid interferon (HLBI) and their combination on human renal cel carcinoma cell line (SMK-R2). Each of VBL, MTX or HLBI tended to suppress [3H]-TdR uptake in a dose-dependent manner. The combination of VBL (0.05 microgram/ml) and HLBI (10(2) or 10(3) IU/ml) produced a subadditive effect, and that of MTX (0.1 microgram/ml) and HLBI 10(2) IU/ml produced a synergistic effect on the human renal cell carcinoma cell line, the effect which is evaluated by Valeriote and Lin's criteria of combination. In particular, the synergistic effect by MTX and HLBI under the clinically achievable drug concentration seems important, when its clinical application is considered.

Rapid and complete hematopoietic reconstitution was achieved in a child with T cell acute lymphoblastic leukemia who was autografted with peripheral blood stem cells (PBSC). A large number of PBSC was collected by two courses of 3-4 hour-lasting lymphopheresis during early remission induced by the second-line chemotherapy and then cryopreserved in liquid nitrogen. A myeloid progenitor cell dose of 203 X 10(4) CFU-GM/kg body weight was reinfused to the patient following marrow-ablative chemotherapy (MCNU 600 mg/m2, cytosine arabinoside 6 g/m2, etoposide 300 mg/m2, cyclophosphamide 160 mg/kg). Neutrophil count reached 0.5 X 10(9)/l by day + 7 and platelet count reached 20 X 10(9)/l by day + 9. Thereafter, white blood cell count continued to increase and reached a maximum of 38 X 10(9)/l on day + 14. Thus, the rapid recovery of hematopoiesis minimised marrow aplasia-related risks. This approach of stem cell rescue operation can be applied to the treatment of children with cancer, who otherwise have no hope to be cured, as an alternative to bone marrow transplantation.

Though Malignant Fibrous Histiocytoma (MFH) has been regarded as a relatively common neoplasm of the mesenchymal soft tissues, the issue of character is largely unresolvable at present. Our department formerly reported that produced experimental MFH by administering 9, 10-dimethyl-1, 2-benzanthracene (DMBA) in male hamsters. In this study I have examined the fine structure of transplantation and heterotransplantation of MFH. The following results were obtained. 1. Five different types of cells were found in the MFH: fibroblast-like cells, histiocyte-like cells, undifferentiated cells, multinucleated giant cells and xanthomatous cells. 2. In histiocyte-like cells, nonphagocytic and actively phagocytic histiocyte-like cells could be distinguished. 3. The multinucleated giant cells could be found two types, fibroblast-like and histiocyte-like. 4. In transplantation, the histology of the original tumor was retained. 5. In heterotransplantation, the tumor was composed mainly of nonphagocytic histiocyte-like cells and undifferentiated cells. 6. The possibility is suggested that both principal cell types in this tumor may derive from the same undifferentiated stem cells.

In vitro chemosensitivity of urological cancers was assessed by a human tumor colony forming assay (HTCA) and a 3H-thymidine incorporation assay. Primary tumor cells from 160 of 253 (63%) urological cancers showed adequate colony growth (greater than 30 colonies per well), and there was a 57% true positive and 100% true negative rate for predicting clinical response of anticancer agents. On the other hand, cells from 37 of 45 (82%) urologic cancers incorporated a sufficient amount of 3H-thymidine (greater than 300 cpm). However, when the positive control drug (chromomycin A3 100 micrograms/ml) was used for the assay quality control, the successful assay rate of the HTCA (38%) was lower than that of the 3H-thymidine incorporation assay (75%), while there was a significant correlation in drug sensitivities between the two assays. Thus, the 3H-thymidine incorporation assay seemed to be more useful than the HTCA for evaluating the chemosensitivity of urologic cancers.

Chemosensitivity tests to anticancer agents using human tumor clonogenic assay (HTCA), novel dye exclusion method (NDE assay), sub-renal capsule assay (SRCA), and chick embryo method (CE method) were utilized to measure the sensitivity of urological malignancies. Surgical tumor specimens from 67 patients with urological malignancies were subjected to HTCA developed by Hamburger and Salmon. An appreciable growth of colonies was obtained in 20 out of 33 renal cancers, 20 out of 30 urothelial cancers and 1 out of 4 testicular tumors examined and colonial growth adequate for chemosensitivity was obtained in 30 of the 67 patients. More than 70% decrease in the plating efficiency after anticancer drug exposure according to Von Hoff's definition, or more than 1.0 value of the "in vivo -in vitro therapeutic index" in terms of the ratio of IC90 to the peak plasma concentration of the drug tested was defined as susceptible. According to Von Hoff's definition, susceptibility to vinblastine (VBL) and cis-dichlorodiammine platinum (CDDP), was seen in 4 out of 11 patients with renal cancer, in 4 out of 15 patients with urothelial cancer and 1 out of 4 patients with renal cancer, respectively. With adriamycin (ADM) it was seen in 3 out of the 15 patients with urothelial cancer, 2 out of 10 patients with renal cancer and 1 patient with testicular tumor. According to TI, susceptibility to VBL was seen in 3 out of 7 patients with renal cancer, and with CDDP it was seen in 2 out of 12 patients with urothelial cancer, and 1 out of 2 patients with renal cancer.(ABSTRACT TRUNCATED AT 250 WORDS)

A number of recent reports have now shown that hematopoietic stem cells are present in the circulation, and these cells have the potential to restore complete hematopoiesis following marrow aplasia. Peripheral stem cells have been collected by continuous flow leukapheresis during periods of hematopoietic regeneration after 2 to 3 weeks of intensive chemotherapy. Following marrow ablative chemotherapy or chemoradiotherapy, the autologous peripheral stem cells are thawed and infused intravenously. When a sufficient stem cell dose was given, rapid neutrophil and platelet recovery has been confirmed, as compared with bone marrow transplantation. Acute leukemia, malignant lymphoma and solid tumors may benefit from this approach, and long-lasting remission or cure may result in a significant number of patients.

Prostate cancer: Considering the stagnation in chemotherapy of prostate cancer in recent years, the following experiments were carried out to determine their clinical value. Surgical specimens from 6 patients, 2 permanent cell lines (EB 33 and PC 93) originated from human prostate cancer and a tumor line serially transplanted in nude mice (PC-NCC) were subjected to chemosensitivity tests such as human tumor cloning assay (HTCA) and/or in vivo tumor growth curve experiments using nude mice. The possible chemosensitive drugs screened by using surgical specimens and PC-NCC tumor were cisplatinum (CDDP), bleomycin (BLM), 5-FU, vincristine (VCR), adriamycin (ADM) and methotrexate (MTX). Most of these drugs were also judged as "effective" by HTCA using a permanent cell line. The minimal discrepancy among them may lead to the conclusion that an in vitro assay using a cell line can substitute for the assay using surgical specimens which can not be obtained frequently. Partly based on the data obtained a chemotherapy regimen, VPM-CisCF, consisting of VCR, peplomycin, MTX, CDDP, cytosine arabinoside (Ara-C) and 5-FU, was designed. The effectiveness of this regimen was demonstrated experimentally. Testis cancer: Two different lines of experiments were performed. A human testicular cancer serially transplanted in nude mice was repeatedly exposed to CDDP in vivo to obtain hyposensitivity to this drug. The synergistic effect of CDDP and VP-16 was demonstrated in the tumor thus obtained. One of its mechanisms has been suggested by partial accumulation of cancer cells in the G1-S and G2-M phase in which CDDP exerts its potential effect.(ABSTRACT TRUNCATED AT 250 WORDS)

Male Small-obese mice (Small-ob) which derived from a C57BL/6 J-ob/ob mouse colony were examined histopathologically at 13-, 39-, and over 52-week-old. C57BL/6 J-ob mice (?/+: Non-ob, ob/ob: Ob) were studied as controls. In Small-ob mice, plasma glucagon concentration was higher than that of the Ob mice (this difference was highly significant), and serum levels for insulin was within normal limits. Microscopically, hypertrophy and hyperplasia of the islets of Langerhans were found only in the pancreas of Ob mice. The increase in the number of A-cells and the decrease in the number of B-cells were revealed immunohistochemically in the islets of Small-ob mice. These changes were more severe with advance of age. In the aged Small-ob mice, perivascular and periductular cell infiltration were found, but inflammatory change of islet tissue was not confirmed in any animals examined. Diabetic symptoms in Small-ob mice seems to stem from the disparity in insulin/glucagon (I/G) ratio associated with hyperglucagonemia which result from increased number of A-cells of pancreatic islets.

We investigated the effect of the ingredients in serum-free cultures, in which serum was completely replaced by albumin, cholesterol and transferrin, on the growth of murine granulocyte/macrophage progenitor cells (colony forming unit in culture: CFU-C) stimulated by serum-free PWM-SCM and the influences of purified granulocyte-macrophage colony stimulating factor (GM-CSF) in serum-free cultures. The results were as follows: 1) The number of CFU-C colonies reached a peak after 4 days of incubation. 2) A linear relationship was observed between the number of CFU-C colonies and the number of inoculated bone marrow cells. 3) Serum-free cultures could support CFU-C colony growth to the same degree as that in serum-containing cultures. Also, no significant differences were found in the types of colonies grown in both cultures. 4) Bovine serum albumin (BSA) and cholesterol were considered to play the most important roles among the ingredients in serum-free cultures. 5) Purified GM-CSF supported CFU-C colony growth in serum-free cultures and more than a half of the colonies formed were GM-colonies. These results showed the usefulness of our serum-free cultures for studying the granulopoiesis in vitro without the influences of various substances in the serum.

Using an in vitro colony forming assay system, cytotoxic effects of anticancer drugs, adriamycin (ADM) and peplomycin (PEP), and the combined effect of hyperthermia and anticancer drugs on cultivated KK-47 cells were investigated. From the response curves obtained at 42 and 43 degrees C hyperthermia, 20% growth inhibition time (IT20) at 42 and 43 degrees C hyperthermia and 50% growth inhibition time (IT50) at 43 degrees C were calculated. The IT20 and IT50 hyperthermia were combined with a 2-hour treatment of each of the anticancer drugs. When the hyperthermia was combined with various concentrations of ADM ranging from 0.005 to 0.1 microgram/ml, enhanced cell killing effects were obtained at the concentrations of less than 0.02 microgram/ml of ADM, whereas, there was no increase in cell killing effect at the concentrations of more than 0.05 microgram/ml of ADM. The combination of hyperthermia with PEP considerably enhanced the cell killing effects with an increase of PEP concentration.

Bone marrow transplantation (BMT) is now increasingly applied not only to hematologic disorders such as leukemias and aplastic anemia but also to some of solid tumors. We experienced ten cases of bone marrow harvest and six cases of autologous or allogeneic BMT during the period from June, 1986 to August, 1987. For autologous BMT cases, bone marrow cells were harvested and cryopreserved at -196 degrees C in liquid nitrogen tank and colony forming assays were examined to ascertain the colony forming ability of harvested bone marrow stem cells. Recovery of colony forming ability after cryopreservation in colony forming unit in culture and burst forming unit in erythrocyte is about 63 to 73% and well preserved. Among 6 cases undergone BMT, five cases are patients with advanced solid tumors including breast cancer, seminoma, lung cancer, malignant lymphoma and brain tumor and these 5 cases have undergone autologous BMT. Conditioning regimen for each cancer in autologous BMT differs based upon conventional chemotherapy regimen and two to four fold dose of conventionally used chemotherapy regimen were administered and after two or three days of high dose chemotherapy, cryopreserved bone marrow cells were thawed and infused. Peripheral white blood cells and platelets were recovered to more than 1000/microliters and 5 x 10(4)/microliters, respectively, within 21 days after bone marrow cell infusion. Disappearance or decrease in tumor size was observed in all cases except brain tumor even though those cases were advanced ones. After autologous BMT, recurrence occurred within three to five months in three cases.(ABSTRACT TRUNCATED AT 250 WORDS)