TUMOR PROMOTERS AND CELL DIFFERENTIATION

肿瘤促进剂和细胞分化

• 批准号: 3166133
• 负责人: LEILA DIAMOND
• 金额: $ 15.84万
• 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/3166133
• 关键词: 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; membrane activity; monoclonal antibody; mutagen testing; mutagens; neoplastic cell; 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)促癌剂抑制BALB/c 3T3细胞脂肪转化 前脂肪细胞，而它们诱导人类分化 早幼粒白血病细胞系HL-60。这项提议的目标是 阐明PES的生化和分子机制 影响差异化。细胞分化程序的生物学研究 前脂肪细胞以及PES对分化的抑制作用 以及编码某些蛋白质的cdna探针 在分化过程中数量发生变化的产品现已问世。这些 将使用探针来比较其中5个的mRNAs的积累 分化BALB/c 3T3前体脂肪细胞的蛋白质 12-0-十四酰佛波醇-13-乙酸酯(TPA)处理，非分化 细胞。TPA诱导的特定mRNA或蛋白积累的改变 将对其蛋白质产物进行分析，以确定肿瘤促进剂 影响该消息的转录或周转或其可译性。 因为已知启动子通过这些刺激乳酸的产生 细胞显著降低了介质的pH值，可能在某些情况下 病例TPA可能会产生抑制正常转录的培养条件 否则将对该消息的翻译进行调查。在HL-60细胞中， PES与其受体蛋白激酶C(PK-C)的初始相互作用是 随后胞质中的PK-C迅速转运到膜组分。 为了研究这种易位的生物学意义 分化后，适当的免疫学和分子探针将 本实验室制备的或从其他实验室获得的。抗病毒抗体 纯化的PK-C将用于鉴定PK-C及其亚细胞 对照和TPA处理的HL-60细胞及其变异体的定位 可逆地抵抗PES诱导分化。这个 抗体也将用于免疫筛选现有的人类cdna。 PK-C cDNA克隆的表达文库；这些PK-C mRNA的探针将被 用来分析PK-C基因表达的转录调控 亲本和变异型HL-60细胞。关于这两个细胞的拟议研究 系统可以增加我们对初始交互作用的理解 细胞和PE以及可能发生的后续事件的作用 这些化合物对细胞分化的控制。