MODULATION OF CELL GROWTH BY ANTISENSE AND ANTIGENE REAGENTS

该项目的重点有三个方面：(1)表征摄取和 未修饰和修饰的寡核苷酸的胞内加工；(2) 反义和抗基因技术在几种体外模型中的应用 识别细胞增殖/病毒关键事件的系统 复制；(3)研究反义和反义基因的作用 作为体内基因表达调节剂的试剂。L：我们有 表征了修饰低聚糖的摄取是能量依赖的， 至少由一种细胞表面结合蛋白介导的内吞过程。 我们设计了一种新的技术来研究细胞内的IOG摄取 定位，以及与蛋白质和核酸的联系。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 内在化的寡头。(2)我们已经证实c-myc抑制是 对正常和恶性淋巴样细胞及某些Burkitt细胞的细胞抑制作用 一种新的方法可以在体外特异性地抑制淋巴瘤细胞的生长 C-myc反义。我们已经证实，N-myc抑制会导致 由于分化状态的改变而导致的生长减慢 神经外胚层来源的细胞系。(3)我们已经证明了c-myc 反义对几种实体肿瘤特别有效， 包括人和大鼠胶质母细胞瘤。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 序列特异性的、非反义介导的对细胞附着的影响 细胞外基质。(4)我们已经确定了能够 特异性抑制bcr-abl酪氨酸激酶和其他酪氨酸激酶 通过与蛋白质的直接相互作用。(5)我们已经能够 可显著延长动物的存活时间 注射c-myc反义基因处理的肿瘤细胞。同时 反义治疗对正常骨髓细胞无影响。

The focus of this project is three-fold: (1) 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, intracellular 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 some 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 glioblastoma. 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. (4) We have identified sequences capable of specifically inhibiting bcr-abl tyrosine kinase and other tyrosine kinases via direct interaction with the proteins. (5) We have been able to demonstrate significant prolongation of animal survival following injection of tumor cells treated with antisense to c-myc. At the same time such antisense treatment has no effect on normal bone marrow cells.