Molecular Analysis of PTHrP in Vascular Smooth Muscle Cell.

为阐明甲状旁腺激素相关肽（PTHrP）在血管系统中的病理生理作用，有必要研究PTHrP受体介导的血管平滑肌细胞（VSMC）或内皮细胞信号转导的分子机制。在目前的项目中，我们已经初步集中分析PTHrP在各种血管系统疾病中的表达。本实验研究了PTHrP的不同片段对原代培养的影响以及转染PTHrP的细胞系的功能，并根据PTHrP在血管平滑肌细胞中的病理生理作用，探讨了PTHrP反义核酸对血管平滑肌细胞的治疗作用。因此，我们研究了PTHrP及其受体在大鼠模型和人类斑块切除术样本中的表达。PTHrP是调节细胞运动性的关键细胞因子（Arterioscl Thromb Vascul Bio 116：565-575 1996）。 关于我们 PTHrP（1-34）、hPTHrP（1-141）、hPTHrP（100-114）和hPTH（1-34）对胸苷掺入、α（1）Ⅱ型胶原基因表达、细胞内cAMP和[Ca^2+]i水平的影响相似。反义寡核苷酸减少PTHrP mRNA翻译，特异性抑制DNA合成。这些数据推测，外源性添加的hPTH和hPTHrP之间没有显著差异，另一方面，细胞内PTHrP除了具有经典的PTH/PTHrP受体介导的功能之外，可能还具有未知的生物学作用（J Endocrinology 150：359-368 1996）。为了进一步扩大基础研究，PTHrP的过度表达在多种与恶性进展相关的癌症中被发现，而不伴有高钙血症。在大鼠中植入PTHrP表达载体产生细胞系的体内实验设计已经证明了体内肿瘤进展和细胞周围钙化基质的形成（Endocrine J 43：527-535 1996）。此外，反义PTHrP寡核苷酸疗法已被报道为针对产生PTHrP的肿瘤的抗血管生成疗法（Cancer Res 56：77-86，1996）。少

To elucidate the patho-physiological role of parathyroid-hormone-related peptide (PTHrP) in vascular system, it is essential to investigate on the molecular mechanism of PTHrP receptor mediated signal transductionin in vascular smooth muscle cells (VSMC) or endotherial cells. In the present projects, we have initially concentrated the analysis of PTHrP expression in various disease involeved with vascular systems. Next, we have studied the effect of various fragments of PTHrP in primary culture and function of the cell lines transfected PTHrP.Finally, we have try to develope the potentially therapeutic approach using antisense PTHrP,based on the pathophysiological role of PTHrP in VSMC.The restenosis is a major problem after baloon angioplasty. Thus, we examined the expression of PTHrP and its receptor in both rat model and human atherectomy samples. PTHrP is a key cytokine to regulate the cell motility (Arterioscl Thromb Vascul Bio1 16 : 565-575 1996).The action of exogenously added h … More PTHrP (1-34), hPTHrP (1-141), hPTHrP (100-114) and hPTH (1-34) on thymidine incorporation, alpha (1) type II co11agen gene expression, intracellular cAMP and [Ca^<2+>]i level was similar. Antisense oligonucleotides decreased PTHrP mRNA translation specifically inhibited DNA synthesis. These data are speculated that there is no significant difference among exogenously added hPTH and hPTHrP,on the other hand intracellular PTHrP may have a yet unknown biological role, in addition to a classical PTH/PTHrP receptor-mediated function (J Endocrinology 150 : 359-368 1996).To further extend the basic research, we have focused on the critical events of cancer by abnormal expression of PTHrP.PTHrP overexpression have been found in various cancers associated with malignant progression without hypercalcemia. The in vivo experimental design which was implanted with PTHrP expresion vector producing cell lines in rat have demanstrated in vivo tumor progression and caltified matrix formation around the cells (Endocrine J 43 : 527-535 1996). Furthermore, the antisense PTHrP oligo-nucleotide therapy have been reported as a anti-angiogenic therapy against PTHrP producing tumor (Cancer Res 56 : 77-86, 1996).Finally, we acknowledge a support from this grant and a valuable contribution of our post doc fellows and staffs. Less