TIMP-1 REGULATORY FACTORS IN HUMAN PROSTATE CANCER

说明(改编自调查员摘要)：规则 金属蛋白酶组织抑制因子(TIMPs)的表达 提出在防止肿瘤细胞扩散方面发挥开创性作用 和转移。这项拨款申请的主要焦点是 调控TIMP-1表达的信号转导途径的研究 人类前列腺癌。在初步研究中，我们(A)确定了 新型顺式作用增强子元件hte-1，以及假定的消音器元件， Hte-2，位于TIMP-1基因5‘端启动子上游；(B) 发现了一种IL-10反应的新型信号处理因子(STAT7)，它可以 被磷酸化并运输到核，在那里它与hte-1结合 激活TIMP-1表达的元件；(C)克隆了STAT7基因并表达 针对3种不同的STAT7多肽的单抗；(D) 稳定表达STAT7的PC-3ML细胞和2×N.I. PC-3细胞；(E)发现TIMP-1和STAT7都与低度恶性有关 人类前列腺癌。由于STAT7的IL-10激活与 TIMP-1的上调我们假设STAT7蛋白协同作用 与其他信号转导因子一起可能在上调中起关键作用 在体外和体内表达TIMP-1以阻断肿瘤细胞的转移。这个 这项赠款申请的具体目标是：目标1：确定 正负转录调控的协同作用 人TIMP-1启动子的元件(即hte-1和hte-2)。目标2： 白介素10受体(IL-10R)信号转导通路在血管内皮细胞癌中的作用 TIMP-1在人前列腺肿瘤细胞中的表达调控目标3： IL-10对肿瘤生长、侵袭、转移及肿瘤影响的研究 转导rSTAT7基因的PC-3细胞的血管生成。目标4：评估 前列腺癌中STAT7和TIMP-1的共同表达总而言之， 这项赠款申请的总体目的有两个：评估是否有 IL-10应答的STAT7蛋白可通过 与人TIMP-1启动子的独特增强子元件结合；以及 确定STAT7表达在肿瘤发生中的潜在重要性 前列腺癌。

DESCRIPTION (Adapted from the Investigator's Abstract): Regulation of expression of the tissue inhibitors of metalloproteinases (TIMPs) is proposed to play a seminal role in the prevention of tumor cell spreading and metastasis. The major focus of this grant application is to characterize the signal transduction pathway regulating TIMP-1 expression in human prostate cancer. In preliminary studies, we have (a) identified a novel cis-acting enhancer element HTE-1, and a putative silencer element, HTE-2, located upstream of the 5' promoter of the TIMP-1 gene; (b) identified an IL-10 responsive novel signal transacting factor (STAT7) that is phosphorylated and transported to nucleus where it binds the HTE-1 element to activate TIMP-1 expression; (c) cloned the STAT7 gene and raised monoclonal antibodies against 3 different STAT7 peptides; (d) demonstrated that STAT7 is expressed in stably transfected PC-3 ML cells and 2 x N.I. PC-3 cells; (e) found both TIMP-1 and STAT7 are associated with low grade human prostate cancer. Since IL-10 activation of STAT7 correlates with the up regulation of TIMP-1 we hypothesize that the STAT7 protein in cooperation with other signal transduction factors might be critical in up regulating TIMP-1 expression in vitro and in vivo to block tumor cell metastasis. The specific aims of this grant application are: Aim 1: Characterize cooperative interactions of positive and negative transcriptional regulatory elements (i.e., HTE-1 and HTE-2) of the human TIMP-1 promoter. Aim 2: Study the role of the IL-10 receptor (IL-10R) signal transduction pathway in the regulation of TIMP-1 expression in human prostatic tumor cells. Aim 3: Evaluate the influence of IL-10 on growth, invasion, metastasis and tumor angiogenesis utilizing rSTAT7 gene transfected PC-3 cells. Aim 4: Evaluate STAT7 and TIMP-1 co-expression in human prostate cancer. In sum, the overall purpose of this grant application is two-fold: to assess whether an IL-10 responsive STAT7 protein can account for up regulation of TIMP-1 via binding to a unique enhancer element of the human TIMP-1 promoter; and to determine the potential importance of STAT7 expression in the tumorigenesis of prostate cancer.