MATURATION OF HUMAN MYELOID LEUKEMIA

我们正在继续研究磷脂和 钙依赖的蛋白激酶(蛋白激酶C)在细胞成熟中的作用 白血病细胞转化为巨噬细胞。过去一年在许多方面生成的数据 实验室，包括我们自己的实验室，证实了我们在 原用途：蛋白激酶C是佛波二酯受体。 我们已经合成了化学定义的甘油二酯，并证明了它们是 “内源性酚类”有能力：(1)激活 蛋白激酶C；(2)竞争性抑制放射性标记佛波二酯 与可溶性或细胞性佛波二酯受体结合；(3)诱导 白血病细胞分化为巨噬细胞；以及(4)模仿佛波醇 双酯诱导的巨噬细胞内磷蛋白的变化 差异化。生物活性佛波醇双酯和细胞渗透性 甘油二酯可诱导MR=21，52，65， 32P负载的HL-60细胞80千道尔顿。今年，重点将是 从HL-60细胞中分离纯化蛋白激酶C及其抗体 制作。主要的磷蛋白将被提纯和32P氨基 测定了酸图和肽图。 这个项目的长期目标是开始了解髓系 在生化水平上成熟。主要科学学科 涉及的学科包括血液学、肿瘤学和细胞生物学。体外成熟 早幼粒细胞白血病细胞作为骨髓的模型 差异化。这样的研究可能会定义分子缺陷。 这会导致髓系成熟停止，从而导致白血病。这个 研究与健康有关，因为它探索了一种 白血病中受阻的分化。作为诱导者活跃的代理人 细胞成熟将很快在实验中得到临床应用 白血病和其他恶性肿瘤的治疗。(B)

We are continuing to investigate the role of phospholipid and Ca2+-dependent protein kinase (protein kinase C) in the maturation of leukemia cells into macrophages. Data generated over the past year in many laboratories, including our own, confirm the hypothesis put forth in our original application: protein kinase C is the phorbol diester receptor. We have synthesized chemically defined diacylglycerols and shown them to be the "endogenous phorbols" as evidenced by their ability to: (1)\activate protein kinase C; (2)\competitively inhibit radiolabeled phorbol diester binding to the soluble or cellular phorbol diester receptor; (3)\induce leukemic cells to differentiate into macrophages; and (4)\mimic phorbol diester-induced phosphoprotein changes that occur during macrophage differentiation. Biologically active phorbol diesters and cell-permeable diacylglycerols induce phosphorylation of proteins with Mr = 21, 52, 65, and 80 kilodaltons in 32P-loaded HL-60 cells. This year, emphasis will be placed on purification of protein kinase C from HL-60 cells and antibody production. The major phosphoproteins will be purified and 32P amino acids and peptide maps determined. The long-term objective of this project is to begin to understand myeloid maturation at a biochemical level. The major scientific disciplines involved are hematology, oncology, and cell biology. In vitro maturation of promyelocytic leukemia cells serves as a model for bone marrow differentiation. Studies such as these may define the molecular defect which leads to cessation of myeloid maturation and hence, leukemia. The investigation is health-related because it explores a molecular pathway of differentiation which is blocked in leukemia. Agents active as inducers of cell maturation will soon find clinical application in the experimental treatment of leukemia and other malignancies. (B)