Molecular Analysis of PTHrP in Hepatic Regeneration

PTHrP 在肝再生中的分子分析

• 批准号: 09670556
• 负责人: OHTSURU Akira
• 金额: $ 2.37万
• 依托单位: Nagasaki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09670556/
• 关键词: PTHrP; liver regeneration; liver specific promoter; Hepatoma; liver cirrhosis; Transgenic model; PTHrP受容体; 肝細胞癌; 癌化; 血管新生

To elucidate the patho-physiological role of parathyroid-hormone-related peptide (PTHrP) in liver disease it is essential to investigate on the molecular mechanism of PTHrP receptor mediated signal transduction in hepatic parenchemal cells and non-parenchemal cells including endothelial cells. In the present projects we have initially concentrated the analysis of PTHrP expression in various disease (J Pathol 182 : 174-9, J Pathol 187 : 217-22). 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 develop the PTHrP transgenic animal using tissue specific promoter of the liver, analyze the pathophysiological role of PTHrP in liver regeneration.The action of exogenously added hPTHrP (1-34), hPTHrP (1-141), hPTHrP (100-114) and hPTH (1-34) on thymidine incorporation, alpha(l) type II collagen gene expression, intracellular cAMP and [Ca^2^+]i level was similar. Antisense oligonucleotides de … More creased 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 Endocrinol 150 : 359-368).To further extend the basic research, we have focused on the critical events induced by abnormal expression of cytokines including PTHrP escaping from physiological regeneration or apoptosis, which can eventually induce gene instability and abnormal cell proliferation. The experimental design is in vivo rat animal model, which is subcutaneously implanted with pituitary tumor producing PTHrP.Surprisingly, these rats arc developed the liver fibrosis and splenoniegaly in 3 months after tumor cell inoculation, and also showed the significantly increased number of GST-P positive focus.Further studies are now continuing on PTHrP transgenic animal model, which was already, established positive transgene clone.Finally, we acknowledge a support from this grant and a valuable contribution of our post doe fellows and staffs. Less

为了阐明甲状旁腺激素相关肽(PTHrP)在肝脏疾病中的病理生理作用，有必要研究PTHrP受体介导的肝旁细胞和包括内皮细胞在内的非旁细胞信号转导的分子机制。在目前的项目中，我们初步集中分析了PTHrP在各种疾病中的表达(J Pathol 182：174-9，J Pathol 187：217-22)。最后，我们利用肝脏组织特异性启动子建立了PTHrP转基因动物，分析了PTHrP在肝再生中的病理生理作用。外源添加hPTHrP(1-34)、hPTHrP(1-141)、hPTHrP(100-114)和hPTH(1-34)对胸腺嘧啶核苷掺入、α(L)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de…更多的PTHrP mRNA翻译特异性地抑制DNA合成。这些数据推测，外源添加的hPTH和hPTHrP之间没有显著差异，另一方面，细胞内的PTHrP可能具有未知的生物学作用，除了经典的PTH/PTHrP受体介导的功能(J Endocrinol 150：359-368)。为了进一步扩大基础研究，我们集中在包括PTHrP在内的细胞因子的异常表达导致的关键事件上，包括PTHrP逃避生理再生或凋亡，最终导致基因不稳定和细胞异常增殖。实验设计为体内动物模型，将产生PTHrP的垂体瘤移植到大鼠体内，接种肿瘤细胞后3个月内出现肝纤维化和脾肿大，GST-P阳性灶明显增多。目前已建立转基因阳性克隆的PTHrP转基因动物模型的进一步研究仍在继续。最后，我们感谢这笔资金的支持和我们的同事和工作人员的宝贵贡献。较少

项目成果

A.Gabit et.al.: "Expression of parathyroid hormone-related paptide in human gastric tumours." J Pathol. 182. 174-179 (1997)

A.Gabit 等人：“甲状旁腺激素相关肽在人胃肿瘤中的表达。”

Nishihara M, Ohtsuru A, et al.: "Clinicopathological implications of parathyroid hormone-related protein in human colorectal tumors."J Pathol. 187. 217-222 (1999)

Nishihara M、Ohtsuru A 等人：“甲状旁腺激素相关蛋白在人类结直肠肿瘤中的临床病理学意义。”J Pathol。

K.Akino et.al.: "Distribution of parathyroid hormone-related peptide and tis receptor in the saccus vasculosus and choroid plexus in the red stingray." Cell Mol Neurobiology. 18. 361-368 (1998)

K.Akino 等人：“红黄貂鱼血管囊和脉络丛中甲状旁腺激素相关肽和 tis 受体的分布。”

Kawashita Y, Ohtsuru A, et al: "Regression of hepatocellular carcinoma in vitro and in vivo by radio-sensitive suicide gene therapy under the inducible and spatial control of radiation." Human Gene Therapy. (in press). (1999)

Kawashita Y、Ohtsuru A 等人：“在辐射的诱导和空间控制下，通过放射敏感的自杀基因疗法在体外和体内消退肝细胞癌。”

Yasuda M, Ohtsuru A, et al: "Stimulation of angiogenesis by low concentration of hydrogen peroxide and the relation with ets-1 in endothelial cells."Life Science. 64. 249-258 (1999)

Yasuda M、Ohtsuru A 等人：“低浓度过氧化氢刺激血管生成及其与内皮细胞中 ets-1 的关系。”生命科学。