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。 (1)我们将未经修饰的寡聚糖的摄取表征为 能量依赖的内吞过程，至少由一个细胞介导 表面结合蛋白。我们设计了一种新的技术来研究 寡聚摄取、细胞内定位及其与蛋白质的关系 和核酸。这种非侵入性技术将允许亚细胞 随着时间的推移，内在化的寡头的本地化。 (2)我们已证实c-myc抑制对正常和非小细胞肺癌具有细胞抑制作用。 恶性淋巴样细胞和一些Burkitt淋巴瘤细胞可以 具体地说，具有c-myc反义基因的体外生长抑制作用。我们 已证实N-myc抑制会导致生长减慢 仅次于分化状态的变化 神经外胚层来源的细胞系。我们已经证明了这种干扰 转化生长因子和自分泌环对上皮细胞和间充质细胞的膀胱抑制作用 细胞。 (3)我们已经证明，持续的皮下灌流 在体内可以显著影响其靶基因的表达和 复制用DNA或RNA观察到的其他体外现象 不合常理。该模型允许对反义寡核苷酸进行体内测试 药效和毒性。我们已经证明了 鞘内持续灌流模型研究丹参的临床疗效 在一个更相关的临床前体内模型系统中反义。

The focus of this project is three-fold: (1) to characterize uptake and intracellular processing of unmodified and modifies 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. (1) We have characterized the uptake of unmodified oligos as an energy-dependent, endocytic process, mediated by at least one cell surface-binding protein. We have devised a novel technique to study oligo 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 that 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. We have demonstrated that interruption of TGF and autocrine loops is cystostatic for epithelial and mesenchymal cells. (3) We have demonstrated that continuous subcutaneous perfusion of an oligo can significantly affest in vivo its targeted gene expression and reproduces other in vitro phenomena observed with either DNA or RNA antisense. This model permits in vivo testing of antisense oligos for efficacy and toxicity. We have demonstrated the feasibility of continuous perfusion intrathecal model to study the clinical efficacy of antisense in a more relevant pre-clinical in vivo model system.