TIMP-1 REGULATORY FACTORS IN HUMAN PROSTATE CANCER

描述（改编自研究者摘要）： 组织金属蛋白酶抑制剂（TIMPs）的表达是 提出在预防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重点是 表征调节TIMP-1表达的信号转导途径， 人类前列腺癌。 在初步研究中，我们（a）确定了一个 新的顺式作用增强子元件HTE-1，和推定的沉默子元件， HTE-2，位于TIMP-1基因的5'启动子上游;（B） 鉴定了一种IL-10应答性新的信号转导因子（STAT 7）， 被磷酸化并转运到细胞核，在那里它与HTE-1结合 （c）克隆STAT 7基因，并将其转染到p53细胞中， 针对3种不同STAT 7肽的单克隆抗体;（d）证实了 STAT 7在稳定转染的PC-3 ML细胞中表达，2 × N.I. （e）发现TIMP-1和STAT 7均与PC-3细胞的低分化相关。 人类前列腺癌。 由于IL-10对STAT 7的激活与细胞凋亡相关， TIMP-1的上调，我们假设STAT 7蛋白协同 与其他信号转导因子的结合可能是上调 TIMP-1在体外和体内表达以阻断肿瘤细胞转移。 的 这项资助申请的具体目标是：目标1：描述 正、负转录调控的协同作用 元件（即，HTE-1和HTE-2）。 目标二： 研究IL-10受体（IL-10 R）信号转导通路在 TIMP-1在人前列腺肿瘤细胞中表达的调控 目标3： 评价IL-10对肿瘤生长、侵袭、转移的影响 利用rSTAT 7基因转染的PC-3细胞进行血管生成。 目标4：评估 STAT 7和TIMP-1在人前列腺癌中的共表达。 总之， 这项补助金申请的总体目的是双重的：评估是否 IL-10应答性STAT 7蛋白可以通过介导TIMP-1的上调来解释TIMP-1的上调。 与人TIMP-1启动子的独特增强子元件结合;以及 确定STAT 7表达在肿瘤发生中的潜在重要性 前列腺癌的症状

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.