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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

流式细胞术判断儿童癌症恶性程度及抗癌药物筛选研究

基本信息

批准号：
61570601
负责人：
HAYASHI Yutaka
金额：
$ 1.34万
依托单位：
TOHOKU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1986
资助国家：
日本
起止时间：
1986 至 1987
项目状态：
已结题

项目摘要

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.

我们计划用流式细胞术分析来研究儿童恶性肿瘤的恶性潜能和筛选抗癌药物。用3种人神经母细胞瘤细胞系进行MMC、ADR、CDDP抗癌药物的流式细胞DNA筛选试验，并与生长抑制试验筛选试验结果进行比较。流式细胞术具有快速、定量分析细胞损伤的优点。培养的细胞需要围绕细胞周期转动至少一次才能进行流式细胞术分析。因此，在原代培养中促进肿瘤细胞的生长是本分析在临床应用中需要解决的问题。我们发现培养的人骨髓单核细胞培养基中含有一种促进培养的神经母细胞瘤细胞生长的物质。该物质的活性可能与粒细胞-巨噬细胞集落刺激因子有关。我们认为该物质对原代培养细胞的筛选试验非常有用。我们成功生产了针对人神经母细胞瘤细胞系（TN-2,NB-1，GOTO）的单克隆抗体。MWT-31和T-93-13是X-63-Ag8-653小鼠骨髓瘤细胞系与经TN-2免疫的BALB/c小鼠脾细胞融合的产物。MWT-31与神经母细胞瘤细胞和周围神经结合。T-93-13与神经母细胞瘤细胞结合。N-14-4是被NB-1免疫的细胞的产物，这种抗体与神经母细胞瘤细胞和周围神经结合。G-16-2-D是它们用GOTO免疫后的产物，它与神经母细胞瘤细胞结合。添加DB-cAMP或维甲酸后，神经母细胞瘤细胞系中抗MWT-31、N-14-4和G-16-2-D的抗原增加。这些单克隆抗体似乎对神经母细胞瘤的诊断和治疗有用。