TUMOR PROMOTERS AND CELL DIFFERENTIATION

肿瘤促进剂和细胞分化

• 批准号: 3166135
• 负责人: LEILA DIAMOND
• 金额: $ 5.52万
• 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/3166135
• 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 3 T3的脂肪转化 前脂肪细胞，而它们诱导人的分化， 早幼粒细胞白血病细胞系HL-60。 本提案的目的是 阐明PE可以通过生物化学和分子机制 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，已经被 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。 这些 探针将用于比较这些中的5种的mRNA的积累， 在分化的BALB/c 3 T3前脂肪细胞中的蛋白质和在 12-0-十四酰基佛波醇-13-乙酸酯（TPA）处理的非分化型 细胞 TPA诱导的特定mRNA或 将分析其蛋白质产物以确定肿瘤促进剂是否 影响信息的转录或周转或其可译性。 由于已知启动子通过这些酶刺激乳酸产生， 细胞，显着降低介质pH值，在一些可能性， TPA可能会产生抑制适当转录的培养条件 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）最初相互作用是 随后胞浆PK-C快速转运至膜成分。 为了研究这种易位的生物意义， 分化，适当的免疫学和分子探针将被 本实验室制备或从他人处获得。 抗体 纯化的PK-C将用于表征PK-C及其亚细胞 在对照和TPA处理的HL-60细胞中以及在 对PE诱导的分化具有可逆抗性。 的 抗体也将用于免疫筛选现有的人cDNA PK-CcDNA克隆的表达文库;这些PK-CmRNA的探针将 用于分析PK-C基因表达的转录调控， 亲本和变体HL-60细胞。 对这两种细胞的拟议研究 系统可以增加我们对初始相互作用的理解， 细胞和PE以及可能发生的后续事件的作用， 通过这些化合物控制细胞分化。