MODULATION OF CELL GROWTH BY ANTISENSE AND ANTIGENE REAGENTS

这个项目的重点是三个方面：(L)表征吸收和 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橄榄摄取 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正常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， 包括人类和大鼠成胶质细胞。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 序列特异性的、非反义介导的对细胞附着的影响 细胞外基质。

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.