TUMOR PROMOTERS AND CELL DIFFERENTIATION

肿瘤促进剂和细胞分化

• 批准号: 3166134
• 负责人: LEILA DIAMOND
• 金额: $ 16.63万
• 依托单位: WISTAR INSTITUTE
• 依托单位国家: 美国
• 财政年份: 1981
• 资助国家: 美国
• 起止时间: 1981-09-01 至 1991-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3166134
• 关键词: binding proteins; carcinogenesis; cell differentiation; cell growth regulation; cell population study; cell type; chemical carcinogenesis; clone cells; cocarcinogen; environment related neoplasm /cancer; erythroid stem cell; gene expression; genetic transcription; genetic translation; human tissue; laboratory rat; membrane activity; monoclonal antibody; mutagen testing; mutagens; neoplastic cell; nucleic acid hybridization; phorbols; polyamines; radioimmunoassay; radiotracer; spectrometry; tritium; tumor promoters

Phorbol ester (PE) tumor promoters inhibit adipose conversion of BALB/c 3T3 preadipocytes whereas they induce differentiation of the human promyelocytic leukemia cell line, HL-60. The objective of this proposal is to elucidate the biochemical and molecular mechanisms by which PEs can affect differentiation. The biology of the differentiation program of the preadipocytes, and of the inhibition of differentiation by PEs, has been very well-characterized; and cDNA probes that encode some of the proteins that alter in amount during differentiation are now available. These probes will be used to compare the accumulation of the mRNAs for 5 of these proteins in differentiating BALB/c 3T3 preadipocytes and in 12-0-tetradecanoylphorbol-13-acetate (TPA)-treated, non-differentiating cells. TPA-induced alterations in the accumulation of a specific mRNA or its protein product will be analyzed to determine if the tumor promoter affects transcription or turnover of that message or its translatability. Since the promoter is known to stimulate lactic acid production by these cells which markedly decreases the medium pH, the possibility that in some cases TPA may create culture conditions that inhibit proper transcription or translation of the message will be investigated. In HL-60 cells, the initial interaction of PEs with their receptor, protein kinase-C (PK-C), is followed by rapid translocation of cytosolic PK-C to membrane components. To study the biological significance of this translocation in differentiation, the appropriate immunological and molecular probes will be prepared in this laboratory or obtained from others. Antibodies to purified PK-C will be used to characterize PK-C and its subcellular localization in control and TPA-treated HL-60 cells and in variants that are reversibly resistant to the induction of differentiation by PEs. The antibodies will also be used to immunoscreen an existing human cDNA expression library for PK-C cDNA clones; these probes for PK-C mRNA will be used to analyze the transcriptional control of PK-C gene expression in the parental and variant HL-60 cells. The proposed studies with these two cell systems can increase our understanding of the initial interaction between cells and PEs and the role of subsequent events that may occur, in the control of cell differentiation by these compounds.

佛波酯 (PE) 肿瘤促进剂抑制 BALB/c 3T3 的脂肪转化 前脂肪细胞，而它们诱导人类的分化 早幼粒细胞白血病细胞系，HL-60。 该提案的目标是 阐明 PE 可以发挥作用的生化和分子机制 影响分化。 分化程序的生物学 前脂肪细胞以及 PE 对分化的抑制作用已被 非常有特点；和编码一些蛋白质的 cDNA 探针 现在可以使用在分化过程中数量发生变化的成分。 这些 探针将用于比较其中 5 个的 mRNA 积累 分化 BALB/c 3T3 前脂肪细胞和 12-0-十四烷酰佛波醇-13-乙酸酯 (TPA) 处理，非分化 细胞。 TPA 诱导的特定 mRNA 积累的改变或 将分析其蛋白质产物以确定肿瘤促进剂是否 影响该消息的转录或周转或其可翻译性。 由于已知启动子可以通过这些促进乳酸的产生 显着降低培养基 pH 值的细胞，在某些情况下可能 TPA 可能会创造抑制正常转录的培养条件 或消息的翻译将被调查。 在 HL-60 细胞中， PE 与其受体蛋白激酶 C (PK-C) 的初始相互作用是 随后细胞质 PK-C 快速易位至膜成分。 研究这种易位的生物学意义 分化，适当的免疫学和分子探针将是 在本实验室制备或从其他实验室获得。 抗体 纯化的 PK-C 将用于表征 PK-C 及其亚细胞 对照和 TPA 处理的 HL-60 细胞以及变体中的定位 对 PE 诱导分化具有可逆性抵抗。 这 抗体也将用于免疫筛选现有的人类 cDNA PK-C cDNA 克隆表达文库；这些 PK-C mRNA 探针将是 用于分析 PK-C 基因表达的转录控制 亲本和变体 HL-60 细胞。 拟议的这两种细胞的研究 系统可以增加我们对之间最初相互作用的理解 细胞和 PE 以及后续可能发生的事件的作用， 这些化合物控制细胞分化。