MOLECULAR REGULATION OF ANGIOGENIC PHENOTYPE

我的研究团队的长期目标是了解分子 人脑胶质瘤生长调控和肿瘤进展的机制。我们 关注于几种生长因子及其受体的作用 这可能与两个主要的病理特征有直接关系 低级别星形细胞瘤向高级别发展的特征 星形细胞瘤：细胞增殖和血管增殖。近期 研究表明，转化生长的表达增加 因子-α(转化生长因子-α)、碱性成纤维细胞生长因子(BFGF)和 血管内皮生长因子在高级别恶性胶质瘤中的表达 提示这些因素在血管生成中起主导作用。在……里面 在接下来的五年里，我们计划将重点放在这些分子的调节上 三种血管生成因子作为未来分子治疗的基础 临床上有恶性胶质瘤。需要检验的假设是1) 血管生成生长因子的表达与恶性肿瘤的关系 表型，以及2)血管生成生长因子表达的调节 导致恶性表型的调节。具体目标是L) 检测碱性成纤维细胞生长因子、转化生长因子-α和血管内皮生长因子的表达及变化 用聚合酶链式反应、原位杂交和原位杂交技术检测脑胶质瘤和癌旁脑组织 免疫组织化学方法；2)调节碱性成纤维细胞生长因子和 使用三螺旋形成寡核苷酸(TFOS)的血管内皮生长因子，使用体外 细胞培养系统用于检测对生长的抑制、对 通过聚合酶链式反应转录、RNA保护实验及基因表达 蛋白质和裸鼠体内移植系统；3)调节 反义方法对血管内皮生长因子和转化生长因子-α活性的影响 核酶、反义寡核苷酸和反义载体； 检测wtP53蛋白表达和两种蛋白的抗血管生成作用 其他新发现的抗血管生成药物。我们相信分子 这里提出的方法来调节关键的血管生成因子的表达 生长因子碱性成纤维细胞生长因子、转化生长因子-α和血管内皮生长因子将为 TFOS和反义TFOS的分子治疗及治疗进展 未来的特工。

The long-term objective of my research team is to understand the molecular mechanism of growth regulation and tumor progression in human gliomas. We have focused on the role of several growth factors and their receptors that may have direct relationships to the two main pathologic features that characterize the progression of low grade astrocytoma to high grade astrocytoma: cellular proliferation and vascular proliferation. Recent studies have demonstrated increased expression of transforming growth factor-alpha (TGF-alpha), basic fibroblast growth factor (bFGF), and vascular endothelial growth factor (VEGF) in high grade malignant gliomas, suggesting that these factors play a predominant role in angiogenesis. In the next five years, we plan to focus on the molecular modulation of these three angiogenic factors as a basis for future molecular therapy for malignant gliomas in the clinic. The hypotheses to be tested are 1) that expression of angiogenic growth factors correlates with malignant phenotype, and 2) that regulation of angiogenic growth factor expression leads to modulation of malignant phenotype. The specific aims are l) to determine the expression and alteration of bFGF, TGF-alpha and VEGF in glioma and brain adjacent to tumor by PCR, in situ hybridization and immunohistochemical methods; 2) to modulate the expression of bFGF and VEGF using triple helix-forming oligonucleotides (TFOs), using an in vitro cell culture system to test for growth inhibition, inhibition of transcription by PCR, RNA protection assay and expression of the mRNA and protein and an in vivo with nude mice xenograft system; 3) to modulate the activity of VEGF and TGF-alpha by antisense approaches including ribozymes, antisense oligonucleotides, and antisense vectors; and 4) to determine the anti-angiogenic effects of wtp53 protein expression and two other newly discovered anti-angiogenic agents. We believe the molecular approaches proposed here to modulate the expression of critical angiogenic growth factors bFGF, TGF-alpha and VEGF will provide insight into molecular therapy and therapeutic development of these TFOs and antisense agents in the future.