Gap junctional regulation of prostate cancer progression

前列腺癌进展的间隙连接调节

• 批准号: 6969536
• 负责人: MAGNUS EDLUND
• 金额: $ 13.16万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-15 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6969536
• 关键词: androgen receptor; androgens; cell adhesion; cell cell interaction; cell growth regulation; clinical research; focal adhesion kinase; gap junctions; hormone regulation /control mechanism; human tissue; immunoprecipitation; membrane channels; metastasis; neoplasm /cancer classification /staging; neoplasm /cancer invasiveness; neoplastic cell; neoplastic transformation; polymerase chain reaction; prostate neoplasms; protein isoforms; protein protein interaction; proteomics; receptor expression; small interfering RNA; stromal cells; tissue /cell culture

DESCRIPTION (provided by applicant): Developing cancers are actively guided by their microenvironments, and at the same time they influence those surroundings. During metastatic invasion, the stromal cells that surround the cancer cells become a local "invasion field," whose micro-vascularization and extracellular matrices differ from normal. Because changes in microenvironment occur relatively rapidly, they have most often been attributed to secreted (indirect) factors exchanged between the two cell populations. Direct cell-cell communication, however, including cytoplasmic channels known as gap junctions, must not be overlooked as a regulatory mechanism during cancer progression. Recent studies of prostate cancer in our laboratory, together with others' work on several carcinomas, reveal increases in the direct gap junctional connection between cancer and stromal cells during cancer progression, as well as increases in the expression levels of specific connexin protein isoforms (out of which the gap junctions are constructed). Evidence is now accumulating that profiles of connexin proteins are critical for invasive behavior. How connexins are regulated during aging, cancer progression and invasion, and how connexins themselves regulate cell behaviors are the focuses of this proposal. We have found that androgen levels (which change as men age) affect connexin profiles, that these profiles affect cell invasion into stromal cell organoids, and that connexins are able to complex with Focal Adhesion Kinase (FAK), a molecule long-known to regulate cell motility via integrin cell adhesion. We aim: 1) to further characterize regulatory effects of cell co-culture, conditioned media, and androgens on connexin expression profiles in cell lines representing different stages of prostate cancer, 2) to use fusion proteins and siRNA to experimentally adjust expression levels of connexin isoforms, searching for effects on cell phenotype and behavior, and 3) to further identify connexin isoform binding-partners, using immunoprecipitation. Not only will these studies clarify connexin protein and gap junctional roles in prostate epithelial-to-mesenchymal transformation (EMT), but they may also reveal diagnostic, connexin "fingerprints" for each cancer stage, and suggest new candidate targets for therapy aimed at decreasing prostate cancer cell invasion and metastasis. Finally, these studies may provide a long-sought link between andropause and prostate cell behavior-between long-distance, hormonal regulatory mechanisms, and those far less-studied mechanisms that depend upon direct cell-cell contact.

描述（由申请人提供）：正在发展的癌症受到其微环境的积极引导，同时它们也影响周围环境。 在转移性侵袭过程中，癌细胞周围的基质细胞成为局部“侵袭场”，其微血管化和细胞外基质与正常细胞不同。 由于微环境的变化发生相对较快，因此它们通常归因于两个细胞群之间交换的分泌（间接）因子。 然而，直接的细胞间通讯，包括被称为间隙连接的细胞质通道，作为癌症进展过程中的一种调节机制，决不能被忽视。 我们实验室最近对前列腺癌的研究，以及其他人对几种癌症的研究，揭示了在癌症进展过程中癌症和基质细胞之间的直接间隙连接连接的增加，以及特定连接蛋白亚型的表达水平的增加（out其中构造了间隙连接）。 现在越来越多的证据表明，连接蛋白的分布对于入侵行为至关重要。 连接蛋白在衰老、癌症进展和侵袭过程中如何受到调节，以及连接蛋白本身如何调节细胞行为是该提案的重点。 我们发现雄激素水平（随着男性年龄的增长而变化）会影响连接蛋白的分布，这些分布会影响细胞侵入基质细胞类器官，并且连接蛋白能够与粘着斑激酶（FAK）复合，FAK是一种长期以来已知的调节分子通过整合素细胞粘附进行细胞运动。 我们的目标是：1) 进一步表征细胞共培养、条件培养基和雄激素对代表前列腺癌不同阶段的细胞系中连接蛋白表达谱的调节作用，2) 使用融合蛋白和 siRNA 通过实验调整连接蛋白的表达水平同种型，寻找对细胞表型和行为的影响，3) 使用免疫沉淀进一步鉴定连接蛋白同种型结合伙伴。 这些研究不仅将阐明连接蛋白和间隙连接在前列腺上皮间质转化（EMT）中的作用，而且还可能揭示每个癌症阶段的诊断连接蛋白“指纹”，并提出新的候选治疗靶点，旨在减少癌症的发生。前列腺癌细胞的侵袭和转移。 最后，这些研究可能提供了男性更年期与前列腺细胞行为之间长期寻求的联系——长距离的激素调节机制，以及那些依赖于直接细胞与细胞接触的、研究较少的机制之间的联系。