MOLECULAR REGULATION OF ANGIOGENIC PHENOTYPE

我的研究团队的长期目标是了解 人胶质瘤生长调节和肿瘤进展的机制。我们 已经关注了几种生长因子及其受体的作用 可能与两个主要的病理特征有直接关系 低级别星形细胞瘤进展为高级别星形细胞瘤的特征 星形细胞瘤：细胞增殖和血管增殖。最近 研究表明转化生长因子的表达增加 因子-α（TGF-α）、碱性成纤维细胞生长因子（bFGF），和 血管内皮生长因子（VEGF）在高级别恶性胶质瘤中的作用， 表明这些因子在血管生成中起主要作用。在 在接下来的五年里，我们计划专注于这些分子的调节， 三种血管生成因子作为未来分子治疗的基础， 恶性胶质瘤的临床表现。待检验的假设是：1） 血管生成生长因子的表达与恶性肿瘤的发生有关 表型，和2）血管生成生长因子表达调节 导致恶性表型的调节。具体目标是：（1） 检测bFGF、TGF-α和VEGF在正常对照组和正常对照组中的表达及变化。 应用PCR、原位杂交、 免疫组织化学方法; 2）调节bFGF的表达， 使用三重螺旋形成寡核苷酸（TFO），使用体外 细胞培养系统，以测试生长抑制、 通过PCR转录、RNA保护测定和mRNA的表达， 蛋白和裸鼠异种移植系统的体内研究; 3）调节 VEGF和TGF-α活性的反义方法，包括 核酶、反义寡核苷酸和反义载体;以及4） 确定wtp 53蛋白表达的抗血管生成作用， 其他新发现的抗血管生成剂。我们相信分子 本文提出的调节关键血管生成因子表达的方法， 生长因子bFGF，TGF-α和VEGF将提供深入了解 这些TFOs和反义寡核苷酸的分子治疗和治疗进展 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.