Role of Raf Kinase Inhibitory Protein in Prostate Cancer

描述（由申请人提供）：生长信号级联失调是前列腺癌发生和恶性进展的关键步骤。在增殖细胞中信号的主要介质是促分裂原活化蛋白激酶（MAPK），其是高度同源的脯氨酸指导的丝氨酸/苏氨酸激酶的超家族，其参与生长和分化促进信号的转导。Ras/Raf激酶级联导致ERK 1和ERK 2（MAPK的ERK亚家族的两个成员）的活化，是正常细胞生长以及肿瘤过程所需的。该信号级联的激活与前列腺癌进展和雄激素非依赖性相关，因此是重要的治疗靶点。Raf激酶的调节剂之一是Raf激酶抑制蛋白（RKIP）。我们实验室最近的研究表明，RKIP通过与S153的相互作用调节Raf的激活;蛋白激酶C激活剂对S153的磷酸化释放RKIP，使Raf随后激活。我们还表明，RKIP调节有丝分裂进程中的一些细胞类型。最近有报道称RKIP在抑制前列腺癌转移中具有令人兴奋的新生物学功能。具体而言，临床组织样本的免疫组织化学分析显示RKIP在原发性前列腺癌中表达，但在转移性病变中不表达。体内功能研究表明，RKIP的异位表达抑制了在体外的侵袭和在体内的转移性C42人前列腺癌细胞模型的良好特征。相反，降低非转移性LNCaP细胞中RKIP的水平促进其体外侵袭能力。总之，这些研究暗示RKIP是一种转移抑制蛋白。我们预计，识别RKIP信号抑制转移的机制将为了解前列腺癌转移的分子基础提供必要的信息。该提案的目标是将RKIP结构与其作为转移抑制剂的功能相关联，然后使用该信息开发调节或模拟RKIP生物功能的治疗试剂。具体来说，我们计划：1）确定前列腺肿瘤细胞中RKIP作用的靶点; 2）产生Raf-1或RKIP的抑制剂或增强剂。在这些研究中开发的试剂可以与其他肿瘤疗法联合使用，以抑制转移级联反应中的特定步骤。

DESCRIPTION (provided by applicant): Dysregulation of growth signaling cascades is a key step in the initiation and malignant progression of prostate cancers. Among the main mediators of signals in proliferative cells are the mitogen-activated protein kinases (MAPKs), a super-family of highly homologous proline-directed serine/threonine kinases that participate in the transduction of growth and differentiation-promoting signals. The Ras/Raf kinase cascade that leads to activation of ERK1 and ERK2, two members of the ERK subfamily of MAPKs, is required for normal cell growth as well as neoplastic processes. Activation of this signaling cascade is correlated with prostate cancer progression and androgen-independence and thus is an important therapeutic target. One of the modulators of Raf kinase is Raf Kinase Inhibitory Protein (RKIP). Recent studies from our laboratory have shown that RKIP regulates Raf activation via interaction at S153; phosphorylation of S153 by activators of protein kinase C release RKIP, enabling subsequent activation of Raf. We have also shown that RKIP regulates mitotic progression in a number of cell types. An exciting new biologic function for RKIP in the suppression of prostate cancer metastasis has recently been reported. Specifically, immunohistochemical analysis of clinical tissue samples revealed expression of RKIP in primary prostate cancer but not in metastatic lesions. Functional in vivo studies showed that ectopic expression of RKIP suppressed invasion in vitro and metastasis in vivo in the well-characterized metastatic C42 human prostate cancer cell model. Conversely, decreasing the level of RKIP in nonmetastatic LNCaP cells promoted their invasive ability in vitro. Taken together, these studies implicate RKIP as a metastasis suppressor protein. We anticipate that identification of the mechanism by which RKIP signals to suppress metastasis will provide information necessary to understanding the molecular underpinnings of prostate cancer metastasis. The goal of this proposal is to relate RKIP structure to its function as a metastasis suppressor and then use this information to develop therapeutic reagents that modulate or mimic RKIP biologic function. Specifically, we plan to: 1) Identify the targets of RKIP action in prostate tumor cells; and 2) Generate inhibitors or potentiators of Raf-1 or RKIP. The reagents developed in these studies could be used in combination with other tumor therapies to suppress specific steps in the metastatic cascade.