MODULATION OF CELL GROWTH BY ANTISENSE AND ANTIGENE REAGENTS

通过反义和抗原试剂调节细胞生长

• 批准号: 3752349
• 负责人: L M NECKERS
• 金额: --
• 依托单位: DIVISION OF CANCER TREATMENT
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3752349
• 关键词: antineoplastics; antisense nucleic acid; binding proteins; biological models; cell adhesion; cell differentiation; cell growth regulation; drug screening /evaluation; endocytosis; extracellular matrix; gene expression; human tissue; intracellular transport; neoplastic cell; nucleic acid inhibitor; oligonucleotides; protooncogene; tissue /cell culture; virus replication

The focus of this project is three-fold: (l) to characterize uptake and intracellular processing of unmodified and modified oligonucleotides; (2) to utilize antisense and antigene technology in several in vitro model systems to identify critical events in cell proliferation/viral replication; and (3) to study the efficacy of antisense and antigene reagents as in vivo modulators of gene expression. (l) We have characterized the uptake of modified oligos as an energy-dependent, endocytic process, mediated by at least one cell surface-binding protein. We have devised a novel technique to study olig uptake, intrcellular localization, and association with protein and nucleic acids. This non- invasive technique will permit subcellular localization over time of an internalized oligo. (2) We have confirmed that c-myc inhibition is cytostatic for normal and malignant lymphoid cells and sane Burkitt lymphoma cells can be specifically growth-arrested in vitro with a novel c-myc antisense. We have confirmed that N-myc inhibition leads to reduction in growth secondary to alteration in differentiative status of neuroectoderm-derived cell lines. (3) We have demonstrated that c-myc antisense is particularly effective against several solid tumors, including human and rat glioblastana. In solid tumors, the c-myc antisense oligonucleotide has, in addition to its antisense effects, a sequence-specific, non-antisense mediated, effect on cellular attachment to extracellular matrix.

该项目的重点有三个：(l) 描述吸收和 未修饰和修饰寡核苷酸的细胞内加工； (2) 在多种体外模型中利用反义和反基因技术 识别细胞增殖/病毒关键事件的系统 复制； (3) 研究反义和反基因的功效 作为体内基因表达调节剂的试剂。 (l) 我们有 将修饰寡核苷酸的吸收表征为能量依赖性， 由至少一种细胞表面结合蛋白介导的内吞过程。 我们设计了一种新技术来研究细胞内寡核苷酸的摄取 定位以及与蛋白质和核酸的关联。 这个非 侵入性技术将允许随着时间的推移进行亚细胞定位 内化寡核苷酸。 (2) 我们已经证实c-myc抑制作用是 对正常和恶性淋巴细胞具有细胞抑制作用，并保持理智的伯基特 (Burkitt) 淋巴瘤细胞可以在体外用一种新的特异性生长抑制 c-myc反义。 我们已经证实 N-myc 抑制会导致 继发于分化状态改变的生长减少 神经外胚层衍生的细胞系。 (3) 我们证明了c-myc 反义对几种实体瘤特别有效， 包括人和大鼠的胶质母细胞。 在实体瘤中，c-myc 反义寡核苷酸除了具有反义作用外，还具有 序列特异性、非反义介导的对细胞附着的影响 至细胞外基质。