フローサイトメトリーによる小児癌の悪性度の判定と抗癌剤スクリーニングに関する研究

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We projected to study malignant petentiality and to screen anticancer agents for malignant tumors in childhood using flowcytometric analysis. Three kinds of human neuroblastoma cell lines were used for the screening test of anticnacer agents (MMC,ADR,CDDP) by flowcytometric DNA analysis and its results were compared with the screening test by growth inhibition test. Flowcytometric procedure is excellent by its rapidity and quantitative analysis of cell damages. Cultured cells are needed to turn around the cell cycle at least one time for flowcytometric analysis. Accordingly, growth promotion of tumor cells in primary culture was a problem to be solved for the clinical application of this analysis. We found that the medium of cultured human monocytes from bone marrow included a substance which promoted growth of cultured neuroblastoma cells. The activity of this substance is suggested to correlate to granulocyte-macrophage conlony-stimulating factor. We think that this substance is extremely useful for the screening test of primary cultured cells.We suceeded in producing of monoclonal antibodies against human neuroblastoma cell lines (TN-2,NB-1,GOTO). MWT-31 and T-93-13 are products of a cell fusion between X-63-Ag8-653 mouse myeloma cell line and spleen cells froma BALB/c mouse immunized with TN-2. MWT-31 binds to neuroblastoma cells and peripheral nerves. T-93-13 binds to neuroblastoma cells. N-14-4 is a product of those cells immunized with NB-1 and this antibody binds to neuroblastoma cells and peripheral nerves. G-16-2-D is a product of them immunized with GOTO and this binds to neuroblastoma cells. Antigens in neuroblastoma cell lines against MWT-31, N-14-4 and G-16-2-D increase by the addition of DB-cAMP or retinoic acid. These monoclonal antibodies seem to be useful for thediagnosis and treatment of neuroblastoma.